@article{maly_edwards_farin_koester_crosier_2021, title={Assessing puberty in female cheetahs (Acinonyx jubatus) via faecal hormone metabolites and body weight}, ISSN={["1448-5990"]}, DOI={10.1071/RD21169}, abstractNote={With fewer than 7500 cheetahs remaining in the wild, ex situ cheetah populations serve as an insurance policy against extinction and a resource to study species’ biology. This study aimed to identify the age of pubertal onset in ex situ female cheetahs using non-invasive faecal steroid hormone monitoring and body weights. Faecal samples from nine female cheetahs were collected two to three times weekly from 2 to 36 months of age and body weights were recorded every 3 months. Faecal oestrogen metabolites (FOM) and faecal glucocorticoid metabolites (FGM) were analysed using enzyme immunoassays and samples were categorised into 6-month intervals to compare endocrine characteristics. Faecal hormone and body weight data were analysed using generalised linear mixed models. Age was a significant predictor of mean and baseline FOM concentrations, number of FOM peaks, mean and maximum FOM peak concentrations and the number of cycles. Female cheetahs aged 24–30 months exhibited a marked rise in mean FOM concentration and the number of FOM peaks and cycles increased with age until 24–30 months. Females attained adult body weight by 21 months of age. Mean and baseline FGM concentrations were highest at the 0–6 and 12–18 months of age groups and did not follow the same FOM patterns. Based on body weight data, the FOM concentrations and peak patterning, females were considered pubertal from 24 to 30 months of age. Characterisation of cheetah puberty has direct and significant implications for the improvement of management and reproductive success of cheetahs under human care. This information is particularly informative for identifying important windows of development, littermate dispersal and breeding introductions.}, journal={REPRODUCTION FERTILITY AND DEVELOPMENT}, author={Maly, Morgan A. and Edwards, Katie L. and Farin, Charlotte E. and Koester, Diana C. and Crosier, Adrienne E.}, year={2021}, month={Nov} }
 @article{llanes_whisnant_knox_farin_2019, title={Assessment of ovulation synchronization protocols in goats and use of pregnancy specific protein B (PSPB) enzyme linked immunsorbent assay (ELISA) to determine kid number at birth}, volume={67}, ISSN={["1879-0054"]}, DOI={10.1016/j.domaniend.2018.11.002}, abstractNote={The efficacy of several protocols for ovulation synchronization and timed artificial insemination (TAI) in goats was examined. In addition, the relationship between levels of pregnancy specific protein B (PSPB) during gestation assessed with a commercially available ELISA and the number of offspring at birth was determined. In Experiment 1, 70 does were randomized into four treatments: (1) breed by estrus [BBE], (2) 6-d treatment with a new [C6N], (3) once-used [C61], or (4) twice-used Controled Internal Drug Release (CIDR) device [C62)]. BBE does received two 15 mg doses of prostaglandin-F2α (PGF) at a 10-d interval and were bred 12 h after estrus onset. CIDR groups received a CIDR for 6 d with 15 mg PGF given at CIDR removal. TAI was performed 48 h after CIDR removal and does were given 50 μg GnRH. All does were inseminated with a single dose of frozen semen using a non-surgical, transcervical technique. Pregnancy rates for the BBE, C6N, C61 and C62 treatment groups were 39% ± 12%, 64% ± 12%, 77% ± 12% and 57% ± 12%, respectively, and did not differ. Reuse of CIDRs, even with reuse extending for a total of 21 d, was as effective as new CIDRs for synchronization of ovulation. In Experiment 2, 68 does were randomized into four treatments: (1) BBE, (2) C6N, (3) NC.Synch [NCS], (4) modified NCS [NCSM]. The BBE and C6N groups were as described for Experiment 1. The NCS and NCSM groups received 15 mg PGF on Day 1, 50 μg GnRH on Day 8 and 15 mg PGF on Day 15 (NCS) or Day 15.5 (NCSM). Does were bred by TAI at 72 h (NCS) or 60 h (NCSM) after the second PGF injection. All does in the NCS and NCSM groups received 50 μg GnRH at TAI. Pregnancy rates were 53% ± 12%, 30% ± 11%, 50% ± 11% and 41% ± 12% for does in the BBE, C6N, NCS and NCSM group, respectively, and did not differ. In Experiment 3, 62 does pregnant to TAI were bled at Days 48 and 85 post-insemination for PSPB. Data on kid numbers and birth weights were subsequently recorded. At Day 48 of gestation, PSPB levels for does birthing singletons were lower than for does birthing twins or triplets (25.0 ± 0.1a, 28.8 ± 0.1b and 30.7 ± 0b ng/mL, respectively, abP<0.05). At Day 85 of gestation, PSPB levels were progressively greater for does birthing singletons versus twins versus triplets (27.0 ± 0.1a, 28.5 ± 0.1b and 31.6 ± 0c ng/mL, abcP<0.05). In conclusion, PSPB concentrations detected using a commercially available ELISA at Day 48 or 85 of gestation could distinguish does carrying single versus multiple fetuses.}, journal={DOMESTIC ANIMAL ENDOCRINOLOGY}, author={Llanes, A. and Whisnant, C. S. and Knox, W. B. and Farin, C. E.}, year={2019}, month={Apr}, pages={54–62} }
 @article{brown_bray_carlstead_dickey_farin_ange-van heugten_2019, title={Individual and environmental risk factors associated with fecal glucocorticoid metabolite concentrations in zoo-housed Asian and African elephants}, volume={14}, ISSN={["1932-6203"]}, DOI={10.1371/journal.pone.0217326}, abstractNote={A recent large-scale welfare study in North America involving 106 Asian (Elephas maximus) and 131 African (Loxodonta africana) elephants at 64 accredited facilities identified links (i.e., risk factors) between zoo environmental factors and a number of welfare outcomes (stereotypic behavior, ovarian acyclicity, hyperprolactinemia, walking and recumbence, body condition, health status, serum cortisol). For this population of elephants, we used the same epidemiological methods to examine associations between those risk factors and two additional welfare outcomes, mean concentration and individual variability (CV) of fecal glucocorticoid metabolite concentrations (FGM) as indicators of stress. Results indicate that African elephants are more responsive to social stressors than Asians, and that poor joint health is a stress-related welfare problem for Asian, but not African elephants in the North American population. For both species, higher FGM concentrations were associated with zoos located at more northern latitudes, whereas lower FGM concentrations were associated with having free access to indoor/outdoor spaces, and spending more time in managed interactions with staff. Also important for captive management, elephants having diverse enrichment options and belonging to compatible social groups exhibited reduced intra-individual variability in FGM concentrations. Our findings show that aspects of the zoo environment can be potential sources of stress for captive elephants, and that there are management activities that may facilitate coping with zoo conditions. Given species differences in factors that affected FGM, targeted, species-specific management approaches likely are needed to ensure good welfare for all elephants.}, number={9}, journal={PLOS One}, publisher={Cold Spring Harbor Laboratory}, author={Brown, Janine L. and Bray, Jessica D. and Carlstead, Kathy and Dickey, David and Farin, Charlotte and Ange-van Heugten, Kimberly}, editor={Hillmann, EdnaEditor}, year={2019}, month={May}, pages={e0217326} }
 @article{maly_edwards_farin_koester_crosier_2018, title={Assessing puberty in ex situ male cheetahs (Acinonyx jubatus) via fecal hormone metabolites and body weights}, volume={268}, ISSN={["1095-6840"]}, DOI={10.1016/j.ygcen.2018.07.011}, abstractNote={Cheetahs are one of the most heavily studied felid species, with numerous publications on health, disease, and reproductive physiology produced over the last 30 years. Despite this relatively long history of research, there is a paucity of crucial biological data, such as pubertal onset, which has direct and significant applications to improved management of ex situ cheetah populations. This study aimed to determine age of pubertal onset in ex situ male cheetahs using non-invasive fecal steroid hormone monitoring and body weights. Fecal samples from 12 male cheetahs from four institutions were collected 2–3 times weekly from 1 to 42 months of age. Fecal androgen and glucocorticoid metabolites were analyzed using enzyme immunoassays previously validated for use with cheetah feces. Animal body weights were recorded monthly. Fecal hormone and body weight data were analyzed using generalized linear mixed models. Androgen concentrations exhibited an increase to levels similar to those observed in adult males by 18–24 months of age, and males attained adult body weights by 21 months of age. Based on these weight data and the initial increase in androgens toward adult concentrations, males were considered pubertal from 18 to 24 months of age. Glucocorticoid concentrations and amplitude of concentration over baseline were also increased during this period. Knowledge about the physiological changes associated with puberty is useful for management and improving reproductive success of cheetah populations under human care, particularly for determining timing of litter separation from dam, littermate dispersal and when to introduce potential breeding pairs.}, journal={GENERAL AND COMPARATIVE ENDOCRINOLOGY}, author={Maly, Morgan A. and Edwards, Katie L. and Farin, Charlotte E. and Koester, Diana C. and Crosier, Adrienne E.}, year={2018}, month={Nov}, pages={22–33} }
 @article{llanes_knox_farin_2016, title={11 COMPARISON OF CONTROLLED INTERNAL DRUG-RELEASE INSERT–BASED AND PROGESTERONE-FREE METHODS FOR OVULATION SYNCHRONIZATION AND TIMED ARTIFICIAL INSEMINATION OF GOATS}, volume={28}, ISSN={1031-3613}, url={http://dx.doi.org/10.1071/RDV28N2AB11}, DOI={10.1071/RDV28N2AB11}, abstractNote={

A CIDR synchronization program is an important tool for facilitating ovulation synchronization and timed AI (OvSynch-TAI). The objective of this study was to test the efficacy of reusing CIDR for OvSynch-TAI compared with that for a progesterone-free OvSynch-TAI protocol (NCS) or a breed by oestrus detection (ED) control. A total of 87 does were randomised into 1 of 5 treatments: (1) ED (control, n = 18), (2) NCS (n = 18), (3) CIDR6-New (n = 17), (4) CIDR6–1X (n = 17), and (5) CIDR6–2X (n = 17). Does in the ED group received two 15-mg doses of PGF2α at a 10-d interval and were bred 12 h after the onset of oestrus following the second PGF2α injection. The NCS group received 15 mg of PGF2α on Day 0, 50 μg of gonadotropin-releasing hormone on Day 8, 15 mg of PGF2α on Day 15, and 50 μg of gonadotropin-releasing hormone on Day 18, concurrently with TAI. The CIDR groups (new, 1X-used, or 2X-used) received a P4 device for a 6-d period, and 15 mg of PGF2α was administered at CIDR removal. Does were bred 48 h after CIDR removal and were given 50 μg of gonadotropin-releasing hormone at TAI. The CIDR in the CIDR6–1X group were previously in place for 10 days before use, whereas CIDR in the CIDR6–2X group were previously in place for 16 days before use. Reused CIDR were rinsed in a diluted Nolvasan solution, followed by a clean water rinse, allowed to air dry, and stored in a refrigerator until time of use. The experiment was conducted in 2 replicates. Within each replicate, all treatments were scheduled so that does were bred during the same 2-day period, and all does were inseminated with a single dose of frozen semen using a nonsurgical (transcervical) technique. Blood samples were taken daily in all treatment groups to monitor concentrations of serum progesterone until the time of breeding. Pregnancy was determined by ultrasonography at 54 and 85 days of gestation. Data were analysed using GLM procedures of SAS (SAS Institute Inc., Cary, NC, USA). Data for pregnancy rates were analysed with a model that included effects of treatment, replicate, and their interactions. Data for serum progesterone concentrations were analysed with a model that included the effects of treatment, replicate, day, and their interactions. Means were separated by Duncan’s test. Mean progesterone differed (P < 0.001) with treatment (5.3 ± 0.8bc, 3.5 ± 0.8c, 7.0 ± 0.8ab, 7.9 ± 0.8a, 6.2 ± 0.8ab ng mL–1 for ED, NCS, CIDR6-New, CIDR6–1X, and CIDR6–2X, respectively; least squares mean ± standard error of the mean). Pregnancy rates for the ED, NCS, CIDR6-New, CIDR6–1X, and CIDR6–2X treatment groups were 39 ± 11%bc, 22 ± 11%c, 64 ± 12%ab, 77 ± 12%a, and 57 ± 12%ab, respectively. In conclusion, reused CIDR were as effective as new CIDR for attaining satisfactory pregnancy rates. Timed AI using a once-used CIDR was more effective for establishing pregnancy than ED and NCS treatments. The lower pregnancy rates in the ED and NCS groups were associated with lower mean progesterone levels during the Ovsynch treatment period before breeding.
This research was supported by the North Carolina Agricultural Experiment Station.
}, number={2}, journal={Reproduction, Fertility and Development}, publisher={CSIRO Publishing}, author={Llanes, A. and Knox, W. B. and Farin, C. E.}, year={2016}, pages={135} }
 @article{barnwell_farin_ashwell_farmer_galphin_farin_2016, title={Differences in mRNA populations of short and long bovine conceptuses on Day 15 of gestation}, volume={83}, ISSN={["1098-2795"]}, DOI={10.1002/mrd.22640}, abstractNote={SUMMARY}, number={5}, journal={MOLECULAR REPRODUCTION AND DEVELOPMENT}, author={Barnwell, Callie V. and Farin, Peter W. and Ashwell, Christopher M. and Farmer, William T. and Galphin, Samuel P., Jr. and Farin, Charlotte E.}, year={2016}, month={May}, pages={424–441} }
 @article{farin_barnwell_farmer_2015, title={Abnormal offspring syndrome}, DOI={10.1002/9781118833971.ch67}, journal={Bovine Reproduction}, author={Farin, C. E. and Barnwell, C. V. and Farmer, W. T.}, year={2015}, pages={620–638} }
 @article{barnwell_farin_whisnant_alexander_farin_2015, title={Maternal serum progesterone concentration and early conceptus development of bovine embryos produced in vivo or in vitro}, volume={52}, ISSN={["1879-0054"]}, DOI={10.1016/j.domaniend.2015.03.004}, abstractNote={The hormone progesterone is essential for proper embryonic development. The objective of this study was to examine the relationship between recipient serum concentrations of progesterone, at the time of embryo transfer and at conceptus recovery, on conceptus development from in vivo- or in vitro-produced embryos. Embryos were produced in vivo by superovulation of Holstein cows (IVO; n = 17) or in vitro with either serum-containing (IVPS; n = 27) or serum-restricted medium (IVPSR; n = 34). Single grade I blastocysts from each embryo production system were transferred into heifers on day 7 of development. Conceptuses were recovered on day 17 of gestation and classified as complete, degenerated, or no conceptus. Compared with the IVO group, in vitro-produced embryos had more (P = 0.055) degenerated conceptuses (IVO, 0%; IVPS, 18.5%; and IVPSR, 20.6%). There were no differences in progesterone concentrations at the time of transfer when recipients received either male or female embryos (P > 0.05). Progesterone concentrations in recipients receiving in vivo-produced embryos were higher (P < 0.05; 3.74 ± 0.4 ng/mL; least-squares mean ± standard error of the mean) on day 7 compared with those receiving in vitro-produced embryos (IVPS, 2.4 ± 0.2; IVPSR, 2.58 ± 0.3 ng/mL). However, there was no difference in progesterone concentration on day 7 between treatment groups for heifers from which short conceptuses (≤194 mm) were recovered on day 17. In contrast, when longer (>194 mm) conceptuses were recovered on day 17, heifers receiving in vitro-produced embryos had lower (P = 0.05) serum concentrations of progesterone on day 7 compared with those receiving in vivo-produced embryos (IVPS, 2.2 ± 0.5; IVPSR, 2.3 ± 0.5; IVO, 3.9 ± 0.5 ng/mL). In conclusion, differences in autonomy may exist between in vitro- and in vivo-produced embryos during the period of conceptus elongation with in vitro-produced embryos relying more on intrinsic factors to influence elongation.}, journal={DOMESTIC ANIMAL ENDOCRINOLOGY}, author={Barnwell, C. V. and Farin, P. W. and Whisnant, C. S. and Alexander, J. E. and Farin, C. E.}, year={2015}, month={Jul}, pages={75–81} }
 @article{farmer_farin_piedrahita_bischoff_farin_2013, title={Expression of antisense of insulin-like growth factor-2 receptor RNA non-coding (AIRN) during early gestation in cattle}, volume={138}, ISSN={0378-4320}, url={http://dx.doi.org/10.1016/J.ANIREPROSCI.2013.01.009}, DOI={10.1016/j.anireprosci.2013.01.009}, abstractNote={The insulin-like growth factor type 2 receptor (IGF2R) regulates fetal growth by removing IGF2 from circulation. In mice, expression of the Igf2r gene is only imprinted after implantation and is associated with expression of the antisense non-coding (nc)RNA, Airn. The objectives of this study were, first, to determine if bovine AIRN was expressed during developmentally important stages of gestation, and second, to determine if expression of bAIRN was affected by method of embryo production. Control reactions confirmed that sequence verified bAIRN PCR amplicons resulted from RNA within the sample and not from genomic DNA contamination. IGF2R mRNA was expressed in all fetal liver samples at Days 35-55 and 70 of gestation as well as in 8 of 9 Day 15 conceptuses, 10 of 10 Day 18 conceptuses, and in all day 7 blastocyst pools. bAIRN was expressed in all samples of fetal liver at Days 35-55 and 70 of gestation. The proportion of conceptuses that expressed bAIRN increased from 1 of 9 at Day 15 of gestation to 8 of 10 at Day 18 of gestation. No bAIRN was expressed in any blastocyst pools. The relative level of bAIRN was greater (P<0.05) in fetal liver from embryos produced in vivo compared to that from embryos produced in vitro. In summary bAIRN was not expressed in blastocyst-stage embryos, was expressed in an increasing proportion of embryos around the time of maternal recognition of pregnancy and was expressed following implantation. Furthermore, relative levels of bAIRN in bovine fetal liver can be altered by method of embryo production.}, number={1-2}, journal={Animal Reproduction Science}, publisher={Elsevier BV}, author={Farmer, W.T. and Farin, P.W. and Piedrahita, J.A. and Bischoff, S.R. and Farin, C.E.}, year={2013}, month={Apr}, pages={64–73} }
 @article{bowdridge_knox_whisnant_farin_2013, title={NCSynch: A novel, progestagen-free protocol for ovulation synchronization and timed artificial insemination in goats}, volume={110}, ISSN={["1879-0941"]}, DOI={10.1016/j.smallrumres.2012.07.025}, abstractNote={The objective of this study was to compare overall pregnancy success achieved using a combined ovulation synchronization-timed artificial insemination (NCSynch-TAI) protocol with that obtained using estrus synchronization and artificial insemination (AI). Multiparous Boer and Boer-cross does (n = 132) were randomly assigned within age (year 1) or parity (years 2 and 3) to one of two treatments. Control does received 15 mg prostaglandin F (PGF; Lutalyse®) on days 1 and 10 of treatment. Estrus onset was checked twice daily with separately penned, intact bucks (year 1) or a vasectomized buck penned with the does (years 2 and 3). Control does were bred by AI 12 h after estrus onset using frozen semen. NCSynch-TAI does received 15 mg PGF on day 1 of treatment. Does received 50 μg GnRH (Cystorelin®) on day 8, and 15 mg PGF was given on day 15. On day 18, NCSynch-TAI does were appointment bred using frozen semen and received 50 μg GnRH at breeding. Pregnancy status was determined using transabdominal ultrasonagraphy with kidding rates also recorded. In year 1, 13 of 15 control does (87%) were detected in estrus with 8 pregnant to AI for an overall pregnancy rate of 53%. For NCSynch-TAI, 15 does were bred and 11 (73%) became pregnant. In year 2, 24 of 26 controls (92%) were detected in estrus with 19 pregnant to AI (73% overall pregnancy rate). For NCSynch-TAI, 26 does were bred and 20 (77%) became pregnant. In year 3, 21 of 25 controls (84%) were detected in estrus with 8 pregnant to AI (32% overall pregnancy rate). For NCSynch-TAI, 25 does were bred and 14 (56%) became pregnant. Across all years, the overall pregnancy rate for NCSynch-TAI does (45/66, 68%) did not differ (P = 0.075) compared to controls (35/66; 53%). Kidding rates for the two treatments (68% vs. 51%, respectively) also did not differ (P = 0.075). In summary, pregnancy success following the use of NCSynch-TAI was comparable to that obtained when does were bred based on detection of estrus.}, number={1}, journal={SMALL RUMINANT RESEARCH}, author={Bowdridge, E. C. and Knox, W. B. and Whisnant, C. S. and Farin, C. E.}, year={2013}, month={Feb}, pages={42–45} }
 @article{farmer_farin_farin_2012, title={140 CHARACTERIZATION OF THE BOVINE ANTISENSE TO INSULIN-LIKE GROWTH FACTOR TYPE 2 RECEPTOR (bAIRN) NONCODING RNA}, volume={24}, ISSN={1031-3613}, url={http://dx.doi.org/10.1071/RDv24n1Ab140}, DOI={10.1071/RDv24n1Ab140}, abstractNote={
Bovine insulin-like growth factor type 2 receptor (IGF2R) is an imprinted gene whose aberrant expression has been implicated in development of abnormal offspring syndrome. Gene IGF2R is a member of the IGF2R/AIRN imprinted gene cluster located on chromosome 9 in cattle. This imprinting cluster is flanked on the proximal 5′ end by MAS1. In the mouse, the antisense to Igf2r noncoding RNA, Airn, is a 108-kb polyadenylated transcript that regulates imprinted expression of Igf2r and other genes within the Igf2r/Airn cluster in association with implantation. Bovine AIRN (bAIRN) is expressed in post-implantation fetal tissues with expression coincident with imprinted expression of IGF2R. Further characterisation of bAIRN is needed before investigation of its potential role in regulating imprinted expression of IGF2R in cattle can be pursued. Therefore, the objective of this study was to identify the length and key characteristics of the bAIRN ncRNA transcript. The PCR primer sets (n = 26) were designed based on genomic sequences to walk down the predicted bAIRN ncRNA sequence by amplifying key segments located along the transcript. Total RNA, extracted from gestational day 150 bovine fetal liver, was DNase-treated to eliminate contaminating genomic DNA. The DNase-treated RNA was then used to generate cDNA as a template for PCR amplification. A putative promoter region containing the transcriptional start site for bAIRN was identified based on NCBI sequence data. This bAIRN promoter is located 441 bp upstream of differentially methylated region 2 (DMR2) within intron 2 of IGF2R and contains a CAAT-box, GC-box and TATA-box. The PCR primer sets (n = 2) designed to amplify products upstream of the putative promoter failed to produce any PCR amplicons. However, PCR primer sets (n = 6) designed to amplify segments located from 4 to 10 kb downstream of the putative promoter region produced amplicons that were sequence verified to be bAIRN ncRNA. Additional PCR primer sets (n = 3) produced sequence-verified amplicons that were located ∼27, 45 and 52 kb downstream of the putative bAIRN transcription start site. Between 27 and 44 kb downstream of the bAIRN promoter, a series of sequence duplications from MAS1 were identified within the bAIRN ncRNA. The role of these duplicated regions is currently unknown. Three PCR primer sets (n = 3) designed to amplify consecutive regions spanning from 93.4 to 101 kb downstream of the bAIRN promoter failed to produce amplicons, suggesting prior termination of the bAIRN transcript. In addition, a strong putative polyadenylation signal was identified at 65.4 kb downstream from the bAIRN promoter. In summary, the full-length bAIRN ncRNA has been characterised. The bAIRN promoter is located ∼440 bp upstream of DMR2 on the IGF2R antisense strand. Transcription from the bAIRN promoter results in a noncoding RNA with a length of ∼65 kb. The central third of the bAIRN transcript contains a series of sequence duplications from MAS1. The function of these duplications within the bAIRN ncRNA is currently unknown.
This research was supported by the North Carolina Agricultural Experiment Station.
}, number={1}, journal={Reproduction, Fertility and Development}, publisher={CSIRO Publishing}, author={Farmer, W. T. and Farin, P. W. and Farin, C. E.}, year={2012}, pages={182} }
 @article{barnwell_whisnant_farin_alexander_farin_2012, title={80 RECIPIENT SERUM PROGESTERONE CONCENTRATION AND EARLY CONCEPTUS DEVELOPMENT IN BOVINE EMBRYOS PRODUCED IN VIVO OR IN VITRO}, volume={24}, ISSN={1031-3613}, url={http://dx.doi.org/10.1071/RDv24n1Ab80}, DOI={10.1071/RDv24n1Ab80}, abstractNote={
The majority of pregnancy loss in cattle occurs during the first 2 to 3 weeks of pregnancy. This loss can be studied by the transfer of in vivo- and in vitro-produced embryos. The objective of this study was to examine the relationship between recipient serum progesterone levels both at the time of embryo transfer and at conceptus recovery on conceptus development from in vivo- or in vitro-produced embryos. Embryos were produced in vivo by superovulation of Holstein cows (IVO; n = 17) or in vitro with either serum-containing (IVPS; n = 27) or serum-restricted medium (IVPSR; n = 34). Single grade-1 blastocysts from each embryo production system were transferred into heifers at Day 7. Conceptuses were recovered at Day 17 of gestation (number recovered/number transferred: IVO, 11/17; IVPS, 16/27; IVPSR, 18/34) and classified as complete, degenerated, or no conceptus. Recipient serum progesterone concentrations were determined by radioimmunoassay and compared with conceptus development outcomes. Sex of conceptus was determined by PCR using a Y-chromosome specific probe. Data were analysed using Fisher's exact test or ANOVA and Duncan's multiple range test. Compared with the IVO group, in vitro-produced embryos had more (P = 0.055) degenerated conceptuses (IVO, 0%; IVPS, 18.5%; IVPSR, 20.6%). There were no differences (P > 0.05) in serum progesterone concentrations in recipients assigned to different treatments at Day 7. There was also no effect (P > 0.05) of treatment on progesterone levels in recipients with either male or female conceptuses at the time of transfer. Interestingly, heifers in the in vitro treatment groups had lower (P < 0.01) progesterone concentrations at Day 7 when no conceptus was recovered at Day 17 (IVPS, 2.1 ± 0.4 ng mL–1; IVPSR, 2.7 ± 0.4 ng mL–1; Least squares means ± standard error of the mean) compared with the IVO group (4.5 ± 0.6 ng mL–1). There was no difference in progesterone concentration between treatment groups for heifers with shorter conceptuses (≤194 mm). However, when longer (>194 mm) conceptuses were recovered, heifers with in vitro produced embryos had lower (P < 0.05) progesterone levels at Day 7 compared with those with in vivo produced embryos (IVPS, 2.2 ± 0.6 ng mL–1; IVPSR, 2.3 ± 0.5 ng mL–1; IVO, 3.9 ± 0.6 ng mL–1). In summary, serum progesterone concentrations in recipients at the time of transfer of in vivo- or in vitro-produced embryos were associated with conceptus development at Day 17 of gestation.
Research supported by NC State University GAANN Biotechnology Fellowship (C. V. Barnwell) and the College of Veterinary Medicine.
}, number={1}, journal={Reproduction, Fertility and Development}, publisher={CSIRO Publishing}, author={Barnwell, C. V. and Whisnant, C. S. and Farin, C. E. and Alexander, J. E. and Farin, P. W.}, year={2012}, pages={152} }
 @article{bing_pratt-phillips_gillen_farin_2011, title={Undergraduate performance in a domestic animal laboratory taught via distance education}, volume={89}, ISSN={["1525-3163"]}, DOI={10.2527/jas.2010-3114}, abstractNote={The objective of this study was to determine if laboratory modules of an undergraduate animal anatomy course offered in distance education (DistEd) format were as effective as face-to-face (F2F) format in helping students learn. Students (n = 159) completed an anatomy pretest as well as a presurvey to assess prior DistEd experience. Alternating each week, laboratory topics were presented either as F2F or as virtual DistEd laboratories. Two laboratory examinations were administered and included material from both laboratory formats (DistEd and F2F). Questions from the pretest were also included and used to generate the posttest scores. At the end of the semester, students completed a postsurvey to determine if DistEd was a viable alternative to F2F. Student grades on each examination were compared using an ANOVA model that included main effects of presentation method (DistEd, F2F), semester (fall, spring), and their interaction. Learning was evaluated based on the performances of students on pre- and posttests using unpaired t-tests. There was an increase (P < 0.0001) in anatomy post- vs. pretest scores for both semesters, indicative of student learning, although there was no effect of presentation method (F2F or DistEd). On exam 1, students achieved greater scores in fall 2008 (P < 0.0001) on material presented via DistEd compared with that presented as F2F. However, in spring 2009 students scored better on material presented as F2F. There was no effect of presentation method on exam 2 scores for either semester. Based on the postsurvey, 79.3% of students in fall 2008 and 52% of students from spring 2009 agreed that DistEd laboratories were a viable alternative to F2F laboratories. The results of this study support the conclusion that anatomy material can be taught effectively by distance education methods.}, number={1}, journal={JOURNAL OF ANIMAL SCIENCE}, author={Bing, J. and Pratt-Phillips, S. and Gillen, L. -A. and Farin, C. E.}, year={2011}, month={Jan}, pages={297–301} }
 @article{wrench_pinto_klinefelter_dix_flowers_farin_2010, title={Effect of season on fresh and cryopreserved stallion semen}, volume={119}, ISSN={["1873-2232"]}, DOI={10.1016/j.anireprosci.2010.02.007}, abstractNote={The objective of this study was to determine the effect of season on sperm quality variables, expression of the fertility-related protein SP22 and selected mRNA transcripts in fresh and cryopreserved stallion sperm. Four stallions were collected in each of the four seasons: summer, fall, winter and spring. Ejaculates were divided and then evaluated for motility, morphology, SP22 staining and expression of selected mRNAs as either fresh semen samples or cryopreserved samples. A significant interaction between season and cryopreservation status was found for total and progressive sperm motility. RNA yield from sperm was not affected by any variable examined. There was no effect of season or cryopreservation on the relative amounts of mRNA for PGK2, TPX1, TIMP3 or ACTB. There was a tendency (P=0.1) for an effect of stallion on the relative amount of ACTB mRNA. The proportion of sperm immunostained for SP22 over the equatorial segment was affected (P<0.05) by stallion. In addition, there was an interaction (P<0.05) between season and cryopreservation status on the percentage of sperm staining for SP22 on the equatorial segment. The correlation among total motility, progressive motility and SP22 immunostaining was much greater (P<0.05) during the breeding season (March and June) than during the non-breeding season (September and December). Based on data analyzed, semen collected in the Northern Hemisphere between March and June may be best suited for cryopreservation.}, number={3-4}, journal={ANIMAL REPRODUCTION SCIENCE}, author={Wrench, N. and Pinto, C. R. F. and Klinefelter, G. R. and Dix, D. J. and Flowers, W. L. and Farin, C. E.}, year={2010}, month={Jun}, pages={219–227} }
 @article{farin_alexander_farin_2010, title={Expression of messenger RNAs for insulin-like growth factors and their receptors in bovine fetuses at early gestation from embryos produced in vivo or in vitro}, volume={74}, ISSN={["0093-691X"]}, DOI={10.1016/j.theriogenology.2010.05.035}, abstractNote={The objective of this study was to determine the effects of in vitro embryo production on physical development and levels of expression of mRNAs for insulin-like growth factor (IGF) ligands (IGF1, IGF2), their receptors (IGF1R, IGF2R), and IGF binding protein-2 (IGFBP2) in bovine fetuses during early gestation. In vivo embryos were recovered from superovulated Holstein cows. For production of embryos in vitro, Holstein oocytes were matured, fertilized, and subsequently cultured in M199 with 10% serum to 168 hpi. On Day 70 of gestation, fetuses (in vivo, n = 14; in vitro, n = 13) were recovered, serum samples collected, and physical measurements recorded. Semi-quantitative RT-PCR assays were used to determine the levels of expression of mRNAs for IGF1, IGF2, IGF1R, and IGF2R in fetal liver and skeletal muscle. Western blots were used to assess levels of IGFBP2 in fetal serum. Fetal body weight did not differ with treatment; however, production of embryos in vitro was associated with decreased crown-nose length and a tendency for increased paired kidney weight, which became significant when expressed on a per bodyweight basis. There was no effect of treatment on levels of IGFBP2 in fetal serum. Levels of IGF1 mRNA in fetal liver were decreased (P < 0.001) in the in vitro group. Levels of IGF2R mRNA in both liver and skeletal muscle were also decreased (P < 0.01) in fetuses from the in vitro group. In summary, fetuses at Day 70 of gestation from embryos produced in vitro had shortened crown-nose length and increased kidney weight on a per bodyweight basis, as well as decreased expression of mRNAs for IGF1 in liver and IGF2R in both liver and skeletal muscle, compared with fetuses from embryos produced in vivo. In conclusion, in vitro embryo culture was associated with subtle changes in fetal development as well as altered expression of both imprinted and non-imprinted genes.}, number={7}, journal={THERIOGENOLOGY}, author={Farin, C. E. and Alexander, J. E. and Farin, P. W.}, year={2010}, month={Oct}, pages={1288–1295} }
 @article{farin_farmer_farin_2010, title={Pregnancy recognition and abnormal offspring syndrome in cattle}, volume={22}, ISSN={["1448-5990"]}, DOI={10.1071/rd09217}, abstractNote={Development of the post-hatching conceptus in ruminants involves a period of morphological expansion that is driven by complex interactions between the conceptus and its intrauterine environment. As a result of these interactions, endometrial physiology is altered, leading to establishment of the pregnancy and continued development of the placenta. Disruption of normal fetal and placental development can occur when embryos are exposed to manipulations in vitro or when inappropriate endocrine sequencing occurs in vivo during the pre- and peri-implantation periods. The present review addresses the development of the post-hatching bovine conceptus, its interactions with the maternal system and changes in development that can occur as a result of in vivo and in vitro manipulations of the bovine embryo.}, number={1}, journal={REPRODUCTION FERTILITY AND DEVELOPMENT}, author={Farin, C. E. and Farmer, W. T. and Farin, P. W.}, year={2010}, pages={75–87} }
 @article{hicks_farin_2009, title={247 CANDIDATE mRNA REGULATING MEIOTIC RESUMPTION IN BOVINE CUMULUS-OOCYTE COMPLEXES}, volume={21}, ISSN={1031-3613}, url={http://dx.doi.org/10.1071/RDv21n1Ab247}, DOI={10.1071/RDv21n1Ab247}, abstractNote={
When cumulus–oocyte complexes (COC) are cultured with follicle-stimulating hormone (FSH), germinal vesicle breakdown (GVBD) occurs and is transcriptionally dependent. In mice, expression of mRNA for nuclear receptor subfamily 4, member 1 (Nr4A1), and early growth response 1 (Egr1) is reduced when GVBD is blocked by the transcriptional inhibitor DRB. The objectives of this study were to define the period required for transcription initiation in bovine COC, determine the pattern of expression for Nr4A1 and Egr1 mRNAs in bovine COC, and reduce Nr4A1 mRNA expression using small interfering RNA (siRNA) to determine the effect on GVBD. In Experiment 1, pools of COC were randomly assigned to mature in TCM-199 supplemented with 10% estrous cow serum, 1 μg mL–1 estradiol, 200 nm pyruvate, 5 μg mL–1 FSH, and 50 μg mL–1 gentamicin. The DRB (120 μm) was added at 0, 30, 60, 90, 120, 150, or 180 min of culture (n = 27 ± 2 COC/treatment per replication; n = 5 replications). The COC were harvested at 20 h and assessed for meiotic stage. In Experiment 2, COC were randomly assigned to maturation medium with FSH and were harvested at 0, 30, 60, 90, and 180 min of culture. Semiquantitative RT-PCR was used to assess Nr4A1, Egr1, and GAPDH (housekeeper) mRNA levels (n = 39 ± 2 COC/treatment per replication; n = 10 replications). In Experiment 3a, COC were matured for 30 min with either FSH, FSH + DRB, or FSH + 25 nm, 50 nm, or 100 nm siRNA for bovine Nr4A1 (siNr4A1). Relative levels of Nr4A1 mRNA were assessed (58 ± 4 COC/treatment per replication; n = 4 replications). In Experiment 3b, COC were matured 9 h in medium containing either FSH, FSH + DRB, FSH + 50 nm siNr4A1, or FSH + 50 nm nonspecific siRNA (siNS). Oocytes were assessed for meiotic stage (n = 13 ± 1 COC/treatment per replication; n = 4 replications). Data were analyzed by one-way ANOVA and Duncan’s test. For Experiment 1, when DRB was added after the start of culture, progressively more oocytes underwent GVBD (least squares mean ± SEM; 99 ± 7%A, 6 ± 7%E, 33 ± 7%D, 64 ± 7%C, 77 ± 7%B,C, 91 ± 7%A,B, 87 ± 7%A,B, 90 ± 7%A,B for control, 0 + DRB, 30 + DRB, 60 + DRB, 90 + DRB, 120 + DRB, 150 + DRB, 180 + DRB; A,B,C,D,EP < 0.05). Thus, the gene transcription required for GVBD occurred between 0 and 60 min of culture. For Experiment 2, relative levels of Nr4A1 mRNA increased (P < 0.05) at 30 min of culture (100 ± 18% v. 192 ± 18%, for 0 v. 30 min). Relative levels of Egr1 mRNA did not change during culture. For Experiment 3a, the relative expression of Nr4A1 mRNA was decreased in cultures containing siNr4A1 (100 ± 6%A, 32 ± 6%D, 62 ± 6%C, 59 ± 6%C,D, 86 ± 6%B for FSH, FSH + DRB, FSH + 25 nm siNr4A1, FSH + 50 nm siNr4A1, FSH + 100 nm siNr4A1; A,B,C,DP < 0.05). For Experiment 3b, there were no differences in the proportions of COC undergoing GVBD at 9 h of culture in the FSH and FSH + siNS treatments. There was a decreased percentage of GVBD oocytes at 9 h of culture with either DRB or 50 nm siNr4A1 (95 ± 4%A, 4 ± 4%C, 65 ± 4%B, 97 ± 4%A for FSH, FSH + DRB, FSH + 50 nm siNr4A1, FSH + 50 nm siNS; A,B,CP < 0.05). These data are consistent with the hypothesis that Nr4A1 may play a role in regulating GVBD in bovine COC.

This research was supported by USDA-NRI Grant 35205-12810, NIH Grant 1 R03 HD043875, and the North Carolina Agricultural Research Service.

}, number={1}, journal={Reproduction, Fertility and Development}, publisher={CSIRO Publishing}, author={Hicks, J. E. and Farin, C. E.}, year={2009}, pages={221} }
 @article{farin_dowdall_hicks_farin_whisnant_2009, title={SUBCUTANEOUS ADMINISTRATION OF FOLLICLE STIMULATING HORMONE FOR SUPEROVULATION OF HOLSTEIN COWS}, volume={21}, ISSN={["1031-3613"]}, DOI={10.1071/RDv21n1Ab293}, abstractNote={
Follicle stimulating hormone (FSH) is usually administered in a series of intramuscular (IM) injections to induce multiple ovulations for embryo production in cattle and other species. The objective of this study was to determine the superovulatory response of dairy cows to subcutaneous (SC) administration of FSH using a reduced number of injections in combination with a progesterone-releasing device. Eighteen non-lactating Holstein cows initially received 25 mg Prostaglandin F2α IM (PGF; Lutalyse; Pfizer Animal Health, Groton, CT, USA) on Day –7. All cows then received an intravaginal progesterone-releasing device (CIDR-B, 1.38 mg progesterone; Pfizer Animal Health) on Day 0, and 100 μg GnRH IM (Cystorelin; Merial Ltd, USA) on Day 2. Cows were randomly assigned to receive a total of 400 mg (20 mL) of FSH (Folltropin-V; Bioniche Animal Health, USA) either by IM injection (IM Group, n = 9 cows) given at 12 h intervals on Days 4 (60 mg, 60 mg), 5 (55 mg, 55 mg), 6 (45 mg, 45 mg) and 7 (40 mg, 40 mg), or by SC injection (SC Group, n = 9 cows) given at 24 h intervals on Days 4 (140 mg), 5 (140 mg) and Day 6 (120 mg). On Day 7, CIDR-B inserts were removed and cows received two 25 mg PGF IM injections given 12 h apart. Cows were artificially inseminated at 12 and 24 h after standing estrus. Blood samples were obtained from all cows at 0, 2, 4, 8, 12, 24, 36, 48, 60, 72, and 84 h after the first FSH injection for determination of serum FSH concentrations. Ovarian follicles and CL were monitored using ultrasonography on Days 4, 7, and 16. Embryos were recovered non-surgically on Day 16 (7 days after estrus). The effects of treatment on follicular response and embryo yield were analyzed by Wilcoxon test, and the response of cows to treatment was analyzed by chi-square test. The effects of treatment on concentrations of serum FSH were analyzed using ANOVA for repeated measures. There was no effect (P > 0.05) of route of FSH administration on the concentrations of serum FSH at any time point. The superovulatory response of cows to treatment, defined as greater than 2 CL per cow, did not differ (P > 0.05) between the IM (77.8%, 7/9 cows) and SC (88.9%, 8/9 cows) Groups. There was also no difference (P > 0.05) between the IM and SC Groups for the number of 5 to 10 mm follicles prior to FSH treatment (mean ± SEM; 0.6 ± 0.2 v. 0.9 ± 0.4), the total number of follicles after FSH treatment (12.4 ± 1.6 v. 12.7 ± 2.2) or the number of CL at embryo recovery (6.4 ± 1.5 v. 10.4 ± 2.1). Similarly, there were no differences (P > 0.05) between the IM and SC Groups for total number of oocytes/embryos (5.6 ± 2.6 v. 13.0 ± 4.3), transferable embryos (Grade 1, 2, 3; 3.0 ± 1.4 v. 6.1 ± 2.9) or Grade 1 embryos (2.9 ± 1.4 v. 4.3 ± 2.5). In conclusion, administration of FSH using 3 SC injections in combination with a progesterone-releasing device was an effective method for superovulation of Holstein cows.

Supported by USDA Animal Health Formula Funds and the State of North Carolina.

}, number={1}, journal={REPRODUCTION FERTILITY AND DEVELOPMENT}, author={Farin, P. W. and Dowdall, K. M. and Hicks, J. E. and Farin, C. E. and Whisnant, C. S.}, year={2009}, pages={243–244} }
 @article{farmer_farin_bischoff_alexander_piedrahita_farin_2008, title={2 DETECTION OF ANTISENSE TO Igf2r (AIR) RNA IN CATTLE}, volume={20}, ISSN={1031-3613}, url={http://dx.doi.org/10.1071/RDv20n1Ab2}, DOI={10.1071/RDv20n1Ab2}, abstractNote={ The insulin-like growth factor type 2 receptor (Igf2r) regulates fetal growth by removing Igf2 from circulation, thus preventing overgrowth. In mice, expression of the Igf2r gene is imprinted only after implantation and is associated with expression of the antisense non-coding (nc)RNA, Air. In contrast, the human IGF2R gene is not imprinted and AIR ncRNA does not exist. Because it is known that the Igf2r gene is imprinted in cattle, the objectives of this study were to determine if Air ncRNA exists in cattle and, if so, whether bovine Air (bAir) is expressed during both pre- and post-implantation development. For objective 1, primer sets were designed for bAir based on bovine genomic sequence. The primer set, bAir3, was used to amplify a region of bAir corresponding to an antisense segment within intron 1 of Igf2r. Primer set bAir4 amplified a segment of bAir ncRNA corresponding to an antisense region upstream of the 52-untranslated region of Igf2r. Pools of whole-cell RNA were extracted from bovine fetal liver and subjected to DNase treatment, reverse transcription (RT), and PCR. Control RT reactions included RT without superscript and RT without superscript or DNase. Controls confirmed that amplification products resulted from RNA present in the sample and not from genomic DNA contamination. Amplicons were obtained for both the bAir3 and the bAir4 primer sets and were sequence verified, demonstrating that bAir ncRNA does exist in cattle. For objective 2, conceptuses (n = 4; mean � SEM length: 2.8 � 0.3 mm) derived from transfer of frozen-thawed in vivo-produced blastocysts were recovered from cows on Day 15 of gestation and snap-frozen for RNA extraction. Samples of liver from in vivo-produced bovine fetuses recovered at Day 70 of gestation (n = 7) were snap-frozen for RNA extraction. Semi-quantitative RT-PCR assays were performed to assess levels of mRNA for Igf2r and H2a, as well as ncRNA for bAir. All conceptus and fetal liver cDNA samples were run in duplicate within the same assay. Relative RNA expression was calculated as the ratio of band intensities of the RNA of interest to that of H2a. Data for relative RNA expression were analyzed by Student's t-test. H2a and Igf2r mRNAs were expressed in all Day 70 fetal liver and Day 15 conceptus samples. Relative levels of Igf2r did not differ (P = 0.19) with stage of development (0.15 � 0.09 v. 0.36 � 0.12 for Day 70 v. Day 15). bAir ncRNA was expressed in 7 of 7 samples of Day 70 fetal liver, whereas only 1 of 4 conceptuses expressed a faint bAir ncRNA signal based on either the bAir3 or bAir4 primer sets (χ2 = 7.23, P < 0.01). Relative levels of bAir ncRNA were greater (P < 0.001) in Day 70 fetal liver compared to those in Day 15 conceptuses for amplicons bAir3 (0.376 � 0.039 v. 0.028 � 0.051) and bAir4 (0.101 � 0.008 v. 0.003 � 0.010). In conclusion, the antisense ncRNA, Air, does exist in cattle and its relative expression is greatest following implantation. These observations are consistent with murine data and suggest that bAir may be involved in regulating imprinted expression of Igf2r in cattle.
}, number={1}, journal={Reproduction, Fertility and Development}, publisher={CSIRO Publishing}, author={Farmer, W. T. and Farin, P. W. and Bischoff, S. R. and Alexander, J. E. and Piedrahita, J. A. and Farin, C. E.}, year={2008}, pages={81} }
 @article{farin_malarkey_alexander_farin_2008, title={201 LIVER AND KIDNEY FUNCTION IN BOVINE FETUSES FROM EMBRYOS PRODUCED IN VIVO OR IN VITRO}, volume={20}, ISSN={1031-3613}, url={http://dx.doi.org/10.1071/RDv20n1Ab201}, DOI={10.1071/RDv20n1Ab201}, abstractNote={Abnormal offspring syndrome can occur in fetuses and calves resulting from embryos produced in vitro or by nuclear transfer procedures. This study was conducted to determine the effects of in vitro embryo culture on fetal biochemistry profiles and histology of the liver and kidneys during late gestation. Embryos were produced in vivo by using superovulated cows (In Vivo) or in vitro by using a serum-containing culture system (In Vitro) as previously described (Miles et al. 2004 Biol. Reprod. 71, 1919–1926). Single blastocysts from each embryo production system were transferred nonsurgically into heifers. On Day 222 of gestation, fetuses from the In Vivo group (n = 12) and the In Vitro group (n = 12) were recovered in utero. Samples of fetal serum were collected for biochemical analysis. Samples of liver and kidney were prepared for histological evaluation. Stereological methods were used to determine the volume density of hepatocytes as well as kidney glomeruli and kidney tubules. Fetuses from the In Vitro group were heavier (P = 0.03) than those from the InVivo group (17.3 � 1.0 kg and 20.7 � 1.0 kg for InVivo and InVitro, respectively; least squares means � SEM). Liver and paired kidney weights per kilogram of body weight did not differ (P  ≥ 0.10) with treatment (26.4 � 0.6 g kg–1  v. 27.6 � 0.6 g kg–1 and 7.8 � 0.5 g kg–1  v. 9.1 � 0.5 g kg–1 for liver and kidney, respectively). In addition, there was no effect of treatment on the volume densities of hepatocytes, kidney glomeruli, and kidney tubules. However, compared with the In Vivo group, fetuses from the In Vitro group had increased (P  ≤ 0.02) concentrations of blood urea nitrogen (BUN; 13.8 � 1.8 mg dL–1  v. 19.8 � 1.8 mg dL–1) and BUN:creatinine ratio (4.6 � 0.8 v. 7.9 � 0.8). No differences were observed between the In Vivo and In Vitro groups for serum levels of creatinine, total bilirubin, total protein, albumin, alanine aminotransferase, aspartate aminotransferase, gamma-glutamyltransferase, glucose, insulin, and insulin-like growth factor-I. In summary, compared with bovine fetuses from in vivo-produced embryos, fetuses from in vitro-produced embryos had increased body weight, normal liver and kidney morphology, and increased concentrations of BUN during late gestation.
Supported by the State of North Carolina.}, number={1}, journal={Reproduction, Fertility and Development}, publisher={CSIRO Publishing}, author={Farin, P. W. and Malarkey, D. E. and Alexander, J. E. and Farin, C. E.}, year={2008}, pages={180} }
 @article{wrench_pinto_klinefelter_dix_flowers_farin_2008, title={Effect of season on sperm membrane protein 22 and selected mRNAs in fresh and cryopreserved stallion sperm}, volume={107}, ISSN={0378-4320}, url={http://dx.doi.org/10.1016/j.anireprosci.2008.05.134}, DOI={10.1016/j.anireprosci.2008.05.134}, abstractNote={The objective was to investigate expression patterns of proteins in pyriform sperm, a common morphological abnormality in bull sperm. Ejaculates were collected from sexually mature Holstein bulls (n = 3) twice weekly for 10 weeks (pre-thermal insult samples). Testicular temperature was elevated in all bulls by scrotal insulation for 72 consecutive hours during week 2. Total sperm proteins were extracted from pre- and post-thermal insult sperm samples and subjected to two-dimensional gel electrophoresis. Among the protein spots detected, 131 spots were significantly expressed (False Detection Rate < 0.01) with ≥ 2 fold changes between normal and pyriform sperm. Among them, 25 spots with ≥ 4 fold difference in expression patterns were identified using liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS). Expression of several proteins involved in sperm capacitation, sperm–egg interaction and sperm cytoskeletal structure was decreased in pyriform sperm, whereas proteins regulating antioxidant activity, apoptosis and metabolic activity were increased. Contents of reactive oxygen species and ubiquitinated proteins were higher in pyriform sperm. In addition to understanding the molecular basis of functional deficiencies in sperm with specific morphological abnormalities, comparing normal versus morphologically abnormal sperm appeared to be a suitable experimental model for identifying important sperm functional proteins.To our knowledge, this study is the first report on differential expression of proteins in pyriform bovine sperm versus morphologically normal sperm. We report that expression of several proteins involved in sperm capacitation, sperm–egg interaction and sperm cytoskeletal structure was decreased in pyriform sperm, whereas proteins which regulate antioxidant activity, apoptosis and metabolic activity were increased. Contents of reactive oxygen species and ubiquitinated proteins were higher in pyriform sperm. In addition to understanding the molecular basis of functional deficiencies in sperm with specific morphological abnormalities, our results suggest that comparing normal versus morphologically abnormal sperm appeared to be a suitable experimental model for identifying important sperm functional proteins.}, number={3-4}, journal={Animal Reproduction Science}, publisher={Elsevier BV}, author={Wrench, N. and Pinto, C.R.F. and Klinefelter, G.R. and Dix, D.J. and Flowers, W.L. and Farin, C.E.}, year={2008}, month={Sep}, pages={357} }
 @article{whang_hunt_wrench_hockney_farin_tonelli_2007, title={Nonoxynol-9-alpha-cyclodextrin inclusion compound and its application for the controlled release of nonoxynol-9 spermicide}, volume={106}, ISSN={["1097-4628"]}, DOI={10.1002/app.26956}, abstractNote={Abstract}, number={6}, journal={JOURNAL OF APPLIED POLYMER SCIENCE}, author={Whang, Hyun Suk and Hunt, Marcus A. and Wrench, Nicola and Hockney, Jessica E. and Farin, Charlotte E. and Tonelli, Alan E.}, year={2007}, month={Dec}, pages={4104–4109} }
 @article{farin_rodriguez_alexander_hockney_herrick_kennedy-stoskopf_2007, title={The role of transcription in EGF- and FSH-mediated oocyte maturation in vitro}, volume={98}, ISSN={["1873-2232"]}, DOI={10.1016/j.anireprosci.2006.10.007}, abstractNote={Understanding mechanisms responsible for meiotic resumption in mammalian oocytes is critical for the identification of strategies to enhance developmental competence of in vitro-matured oocytes. Improvement of in vitro oocyte maturation systems is dependent on a better understanding of mechanisms that regulate oocyte maturation both in vivo and in vitro as well as on the identification of methods to manipulate the meiotic progression of oocytes matured in vitro in a physiological manner. The purpose of this review is two-fold: first, to examine the mechanisms that underlie the acquisition of oocyte developmental competence and regulation of oocyte maturation in vivo and in vitro; second, to present data examining the role of transcription in mediating the ability of EGF and FSH to induce oocyte maturation in vitro. Results presented support the conclusions that (1) EGF-induced oocyte maturation does not require nascent gene transcription in both mice and domestic cats; (2) FSH requires gene transcription to induce oocyte maturation in both species; (3) EGF must be present in the maturation medium to optimize the effectiveness of FSH to promote oocyte maturation; (4) the mechanism used by FSH to induce oocyte maturation in vitro appears to predominate over that used by EGF when both EGF and FSH are present in maturation medium used for either murine or feline cumulus oocyte complexes.}, number={1-2}, journal={ANIMAL REPRODUCTION SCIENCE}, author={Farin, C. E. and Rodriguez, K. F. and Alexander, J. E. and Hockney, J. E. and Herrick, J. R. and Kennedy-Stoskopf, S.}, year={2007}, month={Mar}, pages={97–112} }
 @article{farin_sommer_rodriguez_petters_alexander_2006, title={324 FUNCTIONAL ANALYSIS USING SHORT-INTERFERING (si)RNA OF TRAM-6, A NOVEL TRANSCRIPT ASSOCIATED WITH OOCYTES MATURATION IN CATTLE}, volume={18}, ISSN={1031-3613}, url={http://dx.doi.org/10.1071/RDv18n2Ab324}, DOI={10.1071/RDv18n2Ab324}, abstractNote={Maturation of cumulus–oocyte complexes (COCs) with gonadotropins requires transcription of new mRNAs. When COCs are cultured with FSH and the transcriptional inhibitor DRB, cumulus expansion and germinal vesicle breakdown (GVBD) are arrested. Differential mRNA display was used to identify a novel transcript associated with maturation of COCs (TRAM-6) that is expressed during early maturation but not when maturation is inhibited by DRB. The objective of this study was to use siRNAs targeted against TRAM-6 mRNA to assess its functional role in oocyte maturation. Exp. 1: Pools of 60–10 bovine COCs (n = 5/trt) were randomly assigned to culture in maturation medium consisting of 0.5 mL TCM-199 with 2.5 μg FSH, 0.5 μg estradiol, and 10% estrous cow serum with or without DRB (120 μM) or with increasing doses of siTRAM-6 (25, 50, or 100 nM). Exp. 2: COC pools (n = 4/trt) were cultured in maturation medium with one of the following treatments: control, 120 μM DRB, 100 nM non-specific siRNA (siNS), or 100 nM siTRAM-6. After 4 h of culture, COC pools were used to assess levels of mRNA for TRAM-6, VEGF (vascular endothelial growth factor), and GAPD (glyceraldehyde-3-phosphate dehydrogenase) by semiquantitative RT-PCR. Exp. 3: COCs were cultured in maturation medium for either 8 h (n = 9 pools/trt) or 20 h (n = 7 pools/trt) with one of the following treatments: control, 120 µM DRB, 100 nM siNS, or 100 nM siTRAM-6. Cumulus expansion was graded every 4 h. At the end of culture, a subset of COCs from each treatment was used to assess oocyte meiotic stage. Remaining COCs were used to assess TRAM-6, VEGF, and GAPD mRNA. Data were analyzed using ANOVA and Duncan's test. At 4 h of culture, relative expression of TRAM-6:GAPD mRNA (least squares means ± SEM) was decreased in the DRB and the 100 nM siTRAM-6 treatments but was unaffected by 25 nM TRAM-6, 50 nM siTRAM-6, or 100 nM siNS (Exp. 1: 100 ± 13%a, 17 ± 13%b, 101 ± 13%a, 60 ± 13%ab and 48 ± 13%b for control, DRB, 25 nM siTRAM-6, 50 nM siTRAM-6, and 100 nM siTRAM-6, respectively; abcP < 0.05; Exp. 2: 100 ± 10%a, 14 ± 10%b, 106 ± 10%a and 68 ± 10%c for control, DRB, 100 nM siNS, and 100 nM siTRAM-6, respectively; abcP < 0.05). There was no effect of treatment on expression of VEGF and GAPD mRNA, both of which are present in COCs at the start of culture and are unrelated to TRAM-6 mRNA. At 8 h, GVBD was inhibited by treatment with DRB and siTRAM-6 (68 ± 8%a, 3 ± 8%b, 72 ± 8%a, and 30 ± 8%c for control, DRB, siNSs and siTRAM-6, respectively; abcP < 0.05). At 20 h, the incidence of metaphase II (MII) oocytes was decreased in the DRB and siTRAM-6 groups (96 ± 8%a, 9 ± 8%b, 94 ± 8%a, and 56 ± 8%c for control, DRB, siNS, and siTRAM-6, respectively; abcP < 0.05). Cumulus expansion was inhibited (P < 0.05) by DRB but was not affected by any other treatment. In summary, TRAM-6 siRNA decreased the expression of TRAM-6 mRNA in bovine COCs at 4 h of culture as well as the proportions of oocytes in GVBD at 8 h and in MII at 20 h of culture, but did not affect cumulus expansion. In conclusion, these data are consistent with the identification of a novel mRNA transcript, TRAM-6, that has a functional role in regulating meiotic maturation in bovine COCs.
This work was supported by USDA Grant #2002–35205–12810.}, number={2}, journal={Reproduction, Fertility and Development}, publisher={CSIRO Publishing}, author={Farin, C. E. and Sommer, J. R. and Rodriguez, K. F. and Petters, R. M. and Alexander, J. E.}, year={2006}, pages={269} }
 @article{farin_piedrahita_farin_2006, title={Errors in development of fetuses and placentas from in vitro-produced bovine embryos}, volume={65}, ISSN={["1879-3231"]}, DOI={10.1016/j.theriogenology.2005.09.022}, abstractNote={In vitro systems for oocyte maturation, fertilization and embryo culture [in vitro production (IVP)] have the potential for more wide-spread use in creative breeding programs for dairy and beef cattle. However, one negative consequence of both IVP and somatic cell nuclear transfer (SCNT) in cattle and other species is that embryos, fetuses, placentas, and offspring can differ significantly in morphology and developmental competence compared with those from embryos produced in vivo. Fetuses and placentas derived from IVP and SCNT embryos may fall within the normal range of development, may have obvious abnormalities such as increased fetal and placental weights, or may have subtle abnormalities such as aberrant development of fetal skeletal muscle, placental blood vessels, and altered metabolism. Failures in physiologic and/or genetic mechanisms essential for proper fetal growth and survival outside of the uterus contribute significantly to pregnancy and neonatal losses. Oversized fetuses are at increased risk of death during parturition and the adverse consequences of severe dystocia may compromise the dam. Collectively, these abnormalities have been referred to as ‘large offspring syndrome’ or ‘large calf syndrome’. Abnormal phenotypes resulting from IVP and SCNT embryos are stochastic in occurrence and they have not been consistently linked to aberrant expression of single genes or specific pathophysiology. Thus, reliable methods of early diagnosis of the condition are not yet available. The objective of this paper is to examine abnormal development of fetuses and placentas resulting from embryos produced using in vitro systems. The term ‘abnormal offspring syndrome (AOS)’ is introduced and a classification system of developmental outcomes is proposed to facilitate research efforts on the mechanisms of the various abnormal phenotypes. We also discuss potential genetic and physiologic mechanisms that may contribute to abnormal phenotypes following transfer of IVP and SCNT embryos.}, number={1}, journal={THERIOGENOLOGY}, author={Farin, PW and Piedrahita, JA and Farin, CE}, year={2006}, month={Jan}, pages={178–191} }
 @article{rodriguez_blomberg_zuelke_miles_alexander_farin_2006, title={Identification of candidate mRNAs associated with gonadotropin-induced maturation of murine cumulus oocyte complexes using serial analysis of gene expression}, volume={27}, ISSN={["1094-8341"]}, DOI={10.1152/physiolgenomics.00309.2005}, abstractNote={In cultured cumulus oocyte complexes (COC), FSH induces gene transcription required for germinal vesicle breakdown (GVBD). Experiments were performed to determine the critical period when gene transcription is required for GVBD and to identify candidate mRNAs involved. Experiment I: murine COC were cultured 4 h in the presence of FSH with 5,6-dichloro-1-β-d-ribofuranosylbenzimidazole (DRB) added at different intervals after the start of culture. COC cultured with FSH underwent GVBD (82 ± 7%). When DRB was added at 0, 5, or 10 min after culture initiation, oocyte maturation was blocked (17 ± 7, 14 ± 6, and 21 ± 6% GVBD, respectively). When DRB was added after 15, 20, or 30 min, progressively more COC underwent GVBD (37 ± 6, 39 ± 6, and 66 ± 6%, respectively). The critical period of transcription required for GVBD occurred between 15 and 30 min after culture initiation. Experiment II: COC were cultured for 25 min in the presence (plusDRB) or absence (minusDRB) of DRB. SAGE libraries were generated from COC RNA of each treatment group. A total of 48,431 and 45,367 tags were sequenced for the plusDRB and minusDRB libraries, respectively. Criteria used to identify transcripts of interest included a total tag count of at least 10 across both libraries and a threefold or greater difference in expression between libraries. Using these criteria, 39 and 27 transcripts were identified as differentially expressed at the P ≤ 0.01 and P ≤ 0.001 levels, respectively. Differentially expressed transcripts were classed into major categories that included cell growth, development, and regulation of gene expression. Differentially expressed transcripts represent candidates potentially involved in regulating maturation of murine COC.}, number={3}, journal={PHYSIOLOGICAL GENOMICS}, author={Rodriguez, K. F. and Blomberg, L. A. and Zuelke, K. A. and Miles, J. R. and Alexander, J. E. and Farin, C. E.}, year={2006}, month={Nov}, pages={318–327} }
 @article{miles_farin_rodriguez_alexander_farin_2005, title={Effects of embryo culture on angiogenesis and morphometry of bovine placentas during early gestation}, volume={73}, ISSN={["1529-7268"]}, DOI={10.1095/biolreprod.105.040808}, abstractNote={Abstract The objective of this study was to determine the effects of undefined and semidefined culture systems for in vitro embryo production on angiogenesis and morphometry of bovine placentas during early gestation. Blastocysts produced in vivo were recovered from superovulated Holstein cows and served as controls. Blastocysts produced in vitro were exposed to either serum-supplemented medium with cumulus cell coculture (in vitro-produced with serum; IVPS) or modified synthetic oviductal fluid medium without serum or coculture (mSOF). Single blastocysts from each production system were transferred into heifers. Fetuses and placentas were recovered on Day 70 of gestation. Cotyledonary tissues were obtained for quantification of vascular endothelial growth factor (VEGF) and peroxisome proliferator-activated receptor-gamma (PPARG) mRNA and protein. Samples of placentomes were prepared for immunocytochemistry and histological analysis. Placentas from the mSOF group were heavier and had the fewest placentomes, least placental fluid, and lowest placental efficiency (fetal weight/placental weight) compared with the in vivo and IVPS groups. There was no effect of embryo culture system on volume densities of fetal villi or maternal endometrium within placentomes. The volume density of fetal pyknotic cells was increased in placentomes in the mSOF group compared with the in vivo and IVPS groups. Placentomes in the mSOF group had decreased densities of blood vessels and decreased levels of VEGF mRNA in cotyledonary tissue. In conclusion, compared with placentas from embryos produced in vivo or in vitro using an undefined culture system, placentas from embryos produced in vitro using a semidefined culture system exhibited a greater degree of aberrant development of the placenta during early gestation.}, number={4}, journal={BIOLOGY OF REPRODUCTION}, author={Miles, JR and Farin, CE and Rodriguez, KF and Alexander, JE and Farin, PW}, year={2005}, month={Oct}, pages={663–671} }
 @article{farin_alexander_rodriguez_farin_2004, title={233EXPRESSION OF INSULIN-LIKE GROWTH FACTOR (IGF) MRNA IN BOVINE 
CONCEPTUSES FROM EMBRYOS PRODUCED IN VIVO OR IN VITRO}, volume={16}, ISSN={1031-3613}, url={http://dx.doi.org/10.1071/RDv16n1Ab233}, DOI={10.1071/RDv16n1Ab233}, abstractNote={
The objective of this study was to determine the effect of in vitro embryo production (IVP) on expression of mRNAs for IGF-1, IGF-2, IGF-1 receptor (IGF-1R), IGF-2R and GAPDH in bovine conceptuses at Day 17 of gestation. In vivo embryos (In vivo) were recovered from superovulated Holstein cows. For IVP, Holstein oocytes were matured, fertilized and then cultured in M199 with 10% serum (IVPS) or 1% BSA (IVPSR) to 72hpi. All embryos were then transferred to M199 with 10% serum and cultured to 168hpi. The same Holstein sire was used to produce all embryos. Single grade 1 blastocysts were transferred into recipient heifers. On day 10 after transfer, conceptuses were recovered, measured and stored at −80°C prior to extraction of whole cell (wc) RNA and genomic (g) DNA. wcRNA was assessed for quality based on A260/A280 ratio and visualization of 18S and 28S ribosomal RNA. Only complete (intact) conceptuses with at least 10μg high-quality wcRNA recovered after extraction were included in the analysis (In vivo, n=8; IVPS n=8; IVPSR n=7). Conceptus sex was determined by PCR analysis of gDNA. Semi-quantitative PCR assays were used to assess relative expression of all mRNAs. For each conceptus, 2μg wcRNA were mixed with 3pg rabbit globin mRNA, DNase-treated and reverse-transcribed using random hexamers. Conceptus cDNA samples (140ng each for IGF-1, IGF-2 mRNAs;; 100ng each for IGF-1R, IGF-2R, GAPDH and globin mRNAs) were assayed in duplicate within single assays. Relative mRNA expression was calculated as the ratio of band intensities representing the mRNA of interest to globin mRNA. All data were analyzed for effects of treatment (In vivo, IVPS, IVPSR or In vivo, IVP), sex, stage of blastocyst development at transfer (early, mid, expanded) and the interactions of treatment sex and treatment stage. Conceptus length (LSM±SEMmm) did not differ with treatment (In vivo: 182±55; IVPS: 257±47; IVPSR: 310±72). Expression of GAPDH mRNA was greater (P=0.02) in female conceptuses (1.31±0.11) than in males (0.96±0.07) and did not differ (P=0.14) between treatment groups (In vivo: 1.30±0.11, IVPS: 1.00±0.09, IVPSR: 1.10±0.14). However, when the IVP groups were combined and compared to In vivo controls, expression of GAPDH mRNA was both greater (P=0.002) in females (1.38±0.10) than in males (0.94±0.06) and differed (P=0.03) with treatment (In vivo: 1.30±0.10, IVP: 1.01±0.07). Expression of IGF-1 and IGF-2 mRNA was not detected in the majority of conceptuses. Expression of IGF-1R mRNA was greater (P=0.02) in female (1.19±0.10) than in male conceptuses (0.87±0.07) and differed (P=0.02) with stage of blastocyst development at transfer (early: 0.79±0.07; mid: 1.23±0.13; expanded: 1.07±0.11). There was no effect of treatment on expression of IGF-1R mRNA. Expression of IGF-2R mRNA differed (P=0.03) with treatment (In vivo: 0.85±0.10; IVPS: 0.76±0.09; IVPSR: 0.33±0.13). In conclusion, expression of both imprinted and non-imprinted genes is altered in conceptuses resulting from embryos produced in vitro. Supported by the North Carolina Agricultural Research Service.
}, number={2}, journal={Reproduction, Fertility and Development}, publisher={CSIRO Publishing}, author={Farin, C.E. and Alexander, J.E. and Rodriguez, K.F. and Farin, P.W.}, year={2004}, pages={237} }
 @article{miles_farin_rodriguez_alexander_farin_2004, title={279VASCULAR MORPHOMETRY OF BOVINE PLACENTOMES IN LATE GESTATION FROM 
EMBRYOS PRODUCED IN VIVO OR IN VITRO}, volume={16}, ISSN={1031-3613}, url={http://dx.doi.org/10.1071/RDv16n1Ab279}, DOI={10.1071/RDv16n1Ab279}, abstractNote={
The role of the vascular supply in the development of placentas from embryos produced in vitro is poorly understood. The objective of this study was to determine the effects of in vitro embryo production on morphometry of blood vessels within fetal (cotyledonary) and maternal (caruncular) components of the placentome during late gestation. In vivo-produced embryos were recovered from superovulated Holstein cows on Day 7 after estrus. For in vitro embryo production, oocytes were aspirated from the ovaries of Holstein cows, matured in vitro, and then fertilized. Presumptive zygotes with their cumulus cells were transferred into M-199 with 10% estrus cow serum and cultured for 168h post-insemination. Semen from the same Holstein sire was used for the production of in vivo and in vitro embryos. Single blastocysts from each production system were transferred into the uteri of heifers. On Day 222 of gestation, fetuses and placentas were recovered in utero (in vivo, n=12; in vitro, n=12). Placentomes were collected, fixed and sectioned. Fetal and maternal blood vessels were identified within placentome sections using immunocytochemistry for vascular endothelial growth factor (VEGF) protein. A total of 4.8×105μm2 of tissue were examined from each placentome. Stereological methods were used to determine the volume densities of fetal and maternal blood vessels. Data were analyzed by GLM procedures. Fetuses were heavier (P=0.03) in the in vitro group (20.7±1.0kg, LS mean±SEM) compared to the in vivo group (17.3±1.0kg). Placentas were also heavier (P=0.06) for the in vitro group (2.5±0.2kg) compared to the in vivo group (2.0±0.2kg). Placental efficiency, calculated as fetal weight/placental weight, was similar between the two treatment groups (9.0±0.5 and 8.9±0.5 for in vivo and in vitro, respectively). Fetal vascular volume density in placentomes was not different between the two treatment groups (5.4±0.3% and 5.4±0.3% for in vivo and in vitro, respectively). In contrast, maternal vascular volume density was greater (P=0.02) for placentomes in the in vitro group (5.9±0.3%) compared to in vivo controls (4.9±0.3%). In summary, compared to placentomes from embryos produced in vivo, placentomes from embryos produced in vitro had similar volume density of fetal vessels, but had significantly increased volume density of maternal vessels. Supported by the State of North Carolina.
}, number={2}, journal={Reproduction, Fertility and Development}, publisher={CSIRO Publishing}, author={Miles, J.R. and Farin, C.E. and Rodriguez, K.F. and Alexander, J.E. and Farin, P.W.}, year={2004}, pages={259} }
 @article{dindot_farin_farin_alexander_crosier_walker_long_piedrahita_2004, title={35 Analysis of Epigenetic Modifications and Genomic Imprinting in Nuclear Transfer Derived Bos Gaurus×B. Taurus Concepti}, volume={16}, ISSN={1031-3613}, url={http://dx.doi.org/10.1071/RDv16n1Ab35}, DOI={10.1071/RDv16n1Ab35}, abstractNote={
Somatic cell nuclear transfer in cattle is an inefficient process hindered by low pregnancy rates and fetal placental abnormalities. Improper or incomplete epigenetic reprogramming of the donor genome has been implicated as a cause for these aberrations and has been investigated extensively in mice. Here we report the use of a bovine interspecies model (Bos gaurus×B. taurus) for the assessment and characterization of epigenetic modifications and genomic imprinting in 40-day-old female nuclear transfer (NT)-derived fetuses and placentas. Previously, we identified genomic imprinting at the IGF2, GTL2 and XIST loci in the Bos gaurus×B. taurus fetuses. These results indicated maternal and paternal imprinting of the IGF2 and GTL2 loci, respectively, in the chorion, allantois, liver, lung and brain, whereas the XIST locus was maternally imprinted solely in the chorion of females. We extended this analysis to 40-day-old NT fetuses derived from a hybrid lung fibroblast cell line (female). Analysis of the donor cell line indicated conservation of imprinting of the IGF2 and GTL2 loci and bialleic expression of the XIST locus, presumably from the random patterns of X-chromosome inactivation. Analysis of three NT and three control pregnancies indicated disruption of genomic imprinting at the XIST locus in the chorions of all three clones compared to controls. In contrast, proper allelic expression of the IGF2 and GTL2 loci was observed in all fetuses and placentas. Quantification of maternal and paternal XIST transcripts in the chorion of clones and controls demonstrated a significant skewing from preferential paternal expression in controls (95.0±0.882, mean±S.E.) to mixed paternal and maternal expression in clones (73.6±5.2), (t-test; P<0.05). In an attempt to determine the cause for the abnormal allelic expression of the XIST locus in the chorion of the clones, methylation analysis of the XIST Differentially Methylated Region (DMR) was performed. Methylation-sensitive restriction digests and subsequent PCR of the XIST DMR indicated patterns were not different between controls and clones. However, when genome-wide and promoter-specific methylation analysis (bisulfite sequencing) was extended to the satellite I repeat element and epidermal cytokeratin promoter, hypermethylation was observed in the chorion of clones. These results demonstrate disruption of genomic imprinting in XIST locus in the placenta of 40-day-old clones independent of DMR methylation. They also indicate that cloning is associated with increased levels of methylation in selected genomic regions in the chorion.
}, number={2}, journal={Reproduction, Fertility and Development}, publisher={CSIRO Publishing}, author={Dindot, S.V. and Farin, P. and Farin, C. and Alexander, J. and Crosier, E. and Walker, S. and Long, C. and Piedrahita, J.A.}, year={2004}, pages={140} }
 @article{miles_farin_rodriguez_alexander_farin_2004, title={Angiogenesis and morphometry of bovine placentas in late gestation from embryos produced in vivo or in vitro}, volume={71}, ISSN={["1529-7268"]}, DOI={10.1095/biolreprod.104.031427}, abstractNote={Abstract The objective of this study was to determine the effects of in vitro embryo production on angiogenesis and morphometry of the bovine placenta during late gestation. Blastocysts produced in vivo were recovered from superovulated Holstein cows. Blastocysts produced in vitro were obtained after culture of in vitro-matured and -fertilized Holstein oocytes. Single blastocysts from each production system were transferred into heifers. Fetuses and placentas were recovered on Day 222 of gestation (in vivo, n = 12; in vitro, n = 12). Cotyledonary and caruncular tissues were obtained for quantification of vascular endothelial growth factor (VEGF) and peroxisome proliferator-activated receptor-gamma (PPARγ) mRNA and protein. Tissue sections of placentomes were prepared for morphometric analysis. Fetuses and placentas were heavier from embryos produced in vitro than from embryos produced in vivo. More placentas from embryos produced in vitro had an excessive volume of placental fluid. There was no effect of treatment on the expression of mRNA for VEGF and PPARγ in either cotyledonary or caruncular tissues. The expression of VEGF protein in cotyledons and caruncles as well as the expression of PPARγ protein in cotyledons were not different between the in vitro and in vivo groups. However, caruncles from the in vitro group had increased expression of PPARγ protein. The total surface area of endometrium was greater for the in vitro group compared with controls. In contrast, the percentage placentome surface area was decreased in the in vitro group. Fetal villi and binucleate cell volume densities were decreased in placentomes from embryos produced in vitro. The proportional tissue volume of blood vessels in the maternal caruncles was increased in the in vitro group. Furthermore, the ratios of blood vessel volume density-to-placentome surface area were increased in the in vitro group. In conclusion, these findings are consistent with the concept that compensatory mechanisms exist in the vascular beds of placentas from bovine embryos produced in vitro.}, number={6}, journal={BIOLOGY OF REPRODUCTION}, author={Miles, JR and Farin, CE and Rodriguez, KF and Alexander, JE and Farin, PW}, year={2004}, month={Dec}, pages={1919–1926} }
 @article{rodriguez_farin_2004, title={Developmental capacity of bovine cumulus oocyte complexes after transcriptional inhibition of germinal vesicle breakdown}, volume={61}, ISSN={["1879-3231"]}, DOI={10.1016/j.theriogenology.2003.08.016}, abstractNote={Oocytes cultured in the presence of FSH and the transcriptional inhibitor, 5,6-dichloro-1-beta-d-ribofuranosylbenzimidazole (DRB), remain in meiotic arrest at the germinal vesicle (GV) stage. The objectives of this study were to assess the kinetics of maturation and the developmental capacity of bovine cumulus oocyte complexes (COC) following release from prolonged meiotic arrest by DRB. In Experiment I, COC were cultured for 20 h in Tissue culture medium (TCM)-199 supplemented with 10% estrus cow serum (ECS), 5 microg/ml FSH and 1 microg/ml estradiol in the presence of 120 microM DRB. COC were then released from meiotic arrest and cultured for 20 h in DRB-free medium. CONTROL COC were cultured for 20 h in DRB-free medium, with culture initiated concomitant to the release of DRB-treated COC from meiotic arrest. Nuclear maturation was assessed after 0, 5, 10, 15, and 20 h of culture in DRB-free medium. The proportion of DRB-arrested oocytes reaching metaphase II (MII) following 20 h culture in DRB-free medium was not significantly different from controls ( 96+/-4% versus 99+/-4%). In Experiment II, COC were cultured for 20 h in TCM-199 supplemented with 10% ECS, 10 microg/ml LH, 5 microg/ml FSH, and 1 microg/ml estradiol in the presence or absence of 120 microM DRB. COC in the DRB-treated group were then washed and matured coincident with a second group of control COC for 20 h in DRB-free medium. COC in both groups were fertilized and then randomly assigned to one of two culture systems: TCM-199 + 10%ECS or mSOF + 0.6% fatty acid-free BSA. Development was assessed at 72 h post insemination (hpi), 168 hpi (Day 7) and 216 hpi (Day 9). In this experiment, culture with DRB-arrested oocyte maturation at the GV stage (DRB, 85+/-3% GV; CONTROL, 2+/-3% GV; P<0.001 ). Following release from arrest, maturation and fertilization, the proportion of COC that cleaved by 72 hpi was decreased by treatment with DRB (DRB: 78+/-3% versus90+/-3%; P<0.05). However, no effect of DRB was found on the proportion of cleaved zygotes that reached the blastocyst stage on either Day 7 or Day 9 of culture (Day 7: DRB 16+/-2% versus CONTROL, 21+/-2%; Day 9: DRB 23+/-3% versus CONTROL, 31+/-3%). More embryos reached the blastocyst stage in the TCM-199/serum culture system compared to the mSOF/BSA system on both Days 7 and 9 (Day 7: TCM-199, 23+/-2% versus mSOF, 13+/-2%, P<0.05; Day 9: TCM-199, 32+/-3% versus mSOF, 22+/-3%, P<0.05 ). In summary, bovine COC maintained in meiotic arrest for 20 h by culture in the presence of the transcriptional inhibitor DRB retained their capacity to develop to the blastocyst stage after fertilization in vitro.}, number={7-8}, journal={THERIOGENOLOGY}, author={Rodriguez, KF and Farin, CE}, year={2004}, month={May}, pages={1499–1511} }
 @article{dindot_farin_farin_romano_walker_long_piedrahita_2004, title={Epigenetic and genomic imprinting analysis in nuclear transfer derived Bos gaurus/Bos taurus hybrid fetuses}, volume={71}, DOI={10.1095/biolreprod.103.025775}, abstractNote={Abstract Somatic cell nuclear transfer (NT) in cattle is an inefficient process, whereby the production of calves is hindered by low pregnancy rates as well as fetal and placental abnormalities. Interspecies models have been previously used to facilitate the identification of single nucleotide polymorphisms (SNPs) within coding regions of genes to discriminate between parental alleles in the offspring. Here we report the use of a bovine interspecies model (Bos gaurus × Bos taurus) for the assessment and characterization of epigenetic modifications and genomic imprinting in Day 40-old female NT-derived fetuses and placenta. Analysis of NT and control pregnancies indicated disruption of genomic imprinting at the X inactivation-specific transcript (XIST) locus in the chorion, but not the fetus of clones, whereas proper allelic expression of the insulin-like growth factor II (IGF2) and gene trap locus 2 (GTL2) loci was maintained in both the fetus and placenta. Analysis of the XIST differentially methylated region (DMR) in clones indicated normal patterns of methylation; however, bisulfite sequencing of the satellite I repeat element and epidermal cytokeratin promoter indicated hypermethylation in the chorion of clones when compared with controls. No differences were detected in methylation levels in the fetus proper. These results indicate that the nuclear transfer process affects gene expression patterns in the trophectoderm- and inner cell mass-derived tissues to different extents.}, number={2}, journal={Biology of Reproduction}, author={Dindot, S. V. and Farin, P. W. and Farin, C. E. and Romano, J. and Walker, S. and Long, C. and Piedrahita, J. A.}, year={2004}, pages={470–478} }
 @article{rodriguez_farin_2004, title={Gene transcription and regulation of oocyte maturation}, volume={16}, ISSN={["1448-5990"]}, DOI={10.1071/RD03078}, abstractNote={The developmental potential of an embryo is dependent on the developmental potential of the oocyte from which it originates. The process of oocyte maturation is critical for the efficient application of biotechnologies such as in vitro embryo production and mammalian cloning. However, the overall efficiency of in vitro maturation remains low because oocytes matured in vitro have a lower developmental competence than oocytes matured in vivo. Furthermore, oocytes that have been exposed to gonadotropins have greater developmental competence than oocytes matured in the absence of gonadotropins. By understanding the molecular mechanisms underlying gonadotropin-induced maturation, improvement in oocyte maturation technologies may be expected as procedures to manipulate specific factors involved in signalling for resumption of meiosis are identified. The present review will focus on transcriptional mechanisms underlying the maturation of mammalian oocytes in vitro, as well as on the acquisition of oocyte developmental competence. In addition, a working model for the transcriptional control of mammalian oocyte maturation is proposed.}, number={1-2}, journal={REPRODUCTION FERTILITY AND DEVELOPMENT}, author={Rodriguez, KF and Farin, CE}, year={2004}, pages={55–67} }
 @article{farin_miles_farin_2004, title={Pregnancy loss associated with embryo technologies in cattle}, volume={34}, ISBN={0225-9591}, number={1}, journal={Medecin Veterinaire du Quebec}, author={Farin, P. W. and Miles, J. R. and Farin, C. E.}, year={2004}, pages={62} }
 @article{crosier_farin_rodriguez_blondin_alexander_farin_2002, title={Development of skeletal muscle and expression of candidate genes in bovine fetuses from embryos produced in vivo or in vitro}, volume={67}, ISSN={["0006-3363"]}, DOI={10.1095/biolreprod67.2.401}, abstractNote={Abstract The objectives of this study were to determine the effects of in vitro embryo production on histological development and gene expression in the skeletal muscle of bovine fetuses during late gestation. Blastocysts produced in vivo were obtained from superovulated Holstein cows. Blastocysts produced in vitro were obtained from oocytes of Holstein cows that were matured and fertilized in vitro. Single blastocysts were transferred into heifers at a synchronized estrous and fetuses were recovered at Day 222 of gestation (n = 12 each for in vivo and in vitro). Samples of semitendinosus muscle were obtained for histological analysis and assessment of gene expression. Individual muscle sections were stained for the assessment of primary muscle fibers, secondary muscle fibers, or total muscle fibers. Semiquantitative reverse transcription-polymerase chain reaction assays were performed for 5 different candidate genes. The ratio of secondary-to-primary fiber number was greater in fetuses from embryos produced in vitro compared with fetuses from embryos produced in vivo. Similarly, the ratio of secondary-to-primary fiber volume density tended to be greater in fetuses from embryos produced in vitro. The proportional volume of tissue present between myofibrils was greater in fetuses from embryos produced in vitro. The expression of mRNA for myostatin was decreased in skeletal muscle of fetuses in the in vitro group compared with controls. The expression of mRNA for glyceraldehyde-3-phosphate dehydrogenase tended to be increased in skeletal muscle of fetuses in the in vitro treatment group. There was no effect of treatment on the expression of mRNAs for myf-5, myoD, or myogenin. In conclusion, in vitro production of embryos resulted in fetuses with altered development of skeletal muscle fibers. Myostatin was identified as the candidate gene whose expression may contribute to the observed changes in muscle development of these fetuses.}, number={2}, journal={BIOLOGY OF REPRODUCTION}, author={Crosier, AE and Farin, CE and Rodriguez, KF and Blondin, P and Alexander, JE and Farin, PW}, year={2002}, month={Aug}, pages={401–408} }
 @article{rodriguez_petters_crosier_farin_2002, title={Roles of gene transcription and PKA subtype activation in maturation of murine oocytes}, volume={123}, DOI={10.1530/rep.0.1230799}, abstractNote={The aims of this study were to examine the role of transcription and the coincident involvement of type I and type II protein kinase A (PKA) in the resumption of meiosis in murine cumulus-oocyte complexes (COCs) using the transcriptional inhibitors 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) and alpha-amanitin. The first series of experiments was designed to: (i) characterize the role of transcription in gonadotrophin-mediated and spontaneous maturation of murine oocytes; (ii) examine the roles of specific gonadotrophins (FSH versus hCG) and cumulus cells in transcriptionally mediated oocyte maturation; and (iii) determine the reversibility of the transcriptional arrest of meiosis. In the presence of FSH, transcriptional inhibitors arrested germinal vesicle breakdown (GVBD) (DRB: 2 +/- 2% and control: 76 +/- 2%; alpha-amanitin: 4 +/- 4% and control: 70 +/- 4%). Furthermore, cumulus cells were required for transcriptional inhibitors to arrest GVBD (DRB with cumulus cells: 0 +/- 15%; DRB without cumulus cells: 94 +/- 13%; alpha-amanitin with cumulus cells: 15 +/- 2%; alpha-amanitin without cumulus cells: 99 +/- 2%). Thus, in mice, FSH-mediated GVBD uses a transcriptional mechanism, which probably occurs within the cumulus cell compartment. In a second series of experiments, the role of transcription in mediating the resumption of meiosis after activation of either type I or type II PKA was examined. Activation of type I PKA in murine COCs resulted in an arrest of GVBD that was independent of a transcriptional event (with DRB: 7 +/- 9% GVBD; without DRB: 11 +/- 9% GVBD). In contrast, activation of type II PKA resulted in a resumption of meiosis, which required the occurrence of gene transcription (with DRB: 12 +/- 9% GVBD; without DRB: 80 +/- 9% GVBD). As FSH binding to cumulus cells activates the PKA second messenger system, our results indicate that, in cultured murine COCs, FSH binding to cumulus cells results in the activation of type II PKA, which, in turn, mediates a downstream transcriptional event required for the initiation of GVBD.}, number={6}, journal={Reproduction (Cambridge, England)}, author={Rodriguez, K. F. and Petters, R. M. and Crosier, A. E. and Farin, C. E.}, year={2002}, pages={799–806} }
 @article{farin_crosier_farin_2001, title={Influence of in vitro systems on embryo survival and fetal development in cattle}, volume={55}, ISSN={["1879-3231"]}, DOI={10.1016/S0093-691X(00)00452-0}, abstractNote={In vitro systems are commonly used for the production of bovine embryos. Comparisons between in vivo and in vitro produced embryos illustrate that the morphology of preimplantation-stage embryos differ significantly, the survival of embryos and fetuses is decreased, the size distributions of the populations of conceptuses and fetuses are altered throughout gestation, and placental development is significantly changed. Taken together these findings indicate that exposure to some in vitro environments during the first 7 days of life can profoundly influence fetal and placental development in cattle. An understanding of how in vitro oocyte maturation, in vitro fertilization, and embryo culture systems influence both fetal and placental development should result in systems that consistently produce normal embryos, fetuses, and calves.}, number={1}, journal={THERIOGENOLOGY}, author={Farin, PW and Crosier, AE and Farin, CE}, year={2001}, month={Jan}, pages={151–170} }
 @article{crosier_farin_dykstra_alexander_farin_2001, title={Ultrastructural morphometry of bovine blastocysts produced in vivo or in vitro}, volume={64}, ISSN={["0006-3363"]}, DOI={10.1095/biolreprod64.5.1375}, abstractNote={Abstract The objective of this study was to compare the ultrastructure of bovine blastocysts produced in vivo or in vitro by using morphometric analysis. Blastocysts produced in vivo (multiple ovulations, MO) were obtained from superovulated Holstein cows. For blastocysts produced in vitro, cumulus-oocyte complexes aspirated from ovaries of Holstein cows were matured and fertilized in vitro. At 20 h postinsemination (hpi), zygotes were distributed into one of three culture media: 1) IVPS (in vitro produced with serum): TCM-199 + 10% estrous cow serum (ECS); 2) IVPSR (in vitro produced with serum restriction): TCM-199 + 1% BSA until 72 hpi, followed by TCM-199 + 10% ECS from 72 to 168 hpi; and 3) mSOF (modified synthetic oviductal fluid): mSOF + 0.6% BSA. At 168 hpi, six or seven grade 1 blastocysts from each of the four treatments (MO, IVPS, IVPSR, and mSOF) were fixed and prepared for transmission electron microscopy. Random micrographs of each blastocyst were used to determine the volume density of cellular components. Overall, as blastocysts progressed in development, the volume densities of cytoplasm and intercellular space decreased (P < 0.05) and the volume densities of mature mitochondria, nuclei, blastocoele, and apoptotic bodies increased (P < 0.05). Across treatments, the proportional volumes of nuclei and inclusion bodies were increased in inner cell mass cells compared with trophectoderm cells for mid- and expanded blastocysts. For blastocysts produced in vitro, the volume density of mitochondria was decreased (P < 0.05) as compared with that of blastocycts produced in vivo. The proportional volume of vacuoles was increased (P < 0.05) in blastocysts from the mSOF treatment as compared with blastocysts produced in vivo. For mid- and expanded blastocysts from all three in vitro treatments, the volume density of lipid increased (P < 0.05) and the volume density of nuclei decreased (P < 0.05) compared with those of blastocysts produced in vivo. In conclusion, blastocysts produced in vitro possessed deviations in volume densities of organelles associated with cellular metabolism as well as deviations associated with altered embryonic differentiation. However, the specific nature of these deviations varied with the type of culture conditions used for in vitro embryo production.}, number={5}, journal={BIOLOGY OF REPRODUCTION}, author={Crosier, AE and Farin, PW and Dykstra, MJ and Alexander, JE and Farin, CE}, year={2001}, month={May}, pages={1375–1385} }
 @article{blondin_farin_crosier_alexander_farin_2000, title={In vitro production of embryos alters levels of insulin-like growth factor-II messenger ribonucleic acid in bovine fetuses 63 days after transfer}, volume={62}, ISSN={["0006-3363"]}, DOI={10.1095/biolreprod62.2.384}, abstractNote={Abstract The objective of this study was to determine the effect of embryo production systems on the expression of insulin-like growth factor (IGF)-II mRNA in fetal bovine tissues at Day 70 of gestation (63 days after transfer). Oocytes aspirated from ovaries of Holstein cows were matured and fertilized in vitro. Zygotes were cultured in either tissue culture medium (TCM)-199 + 10% estrous cow serum (ECS; in vitro-produced with serum [IVPS]) or TCM-199 + 1% BSA (in vitro-produced with serum restriction [IVPSR]). At 72 h postinsemination, IVPSR embryos were transferred into fresh TCM-199 + 10% ECS whereas IVPS embryos had fresh medium replaced. All embryos were cultured for an additional 96 h. In vivo-produced embryos were harvested from superovulated Holstein cows (multiple ovulations [MO]). Grade 1 blastocysts from all groups were transferred singly into Angus heifers. At Day 70 of gestation, fetuses (n = 14, 13, and 11 for MO, IVPS, and IVPSR, respectively) were collected; liver and skeletal muscle samples were snap frozen, and whole-cell RNA (wcRNA) was extracted. Levels of IGF-II mRNA were determined by RNase protection assay and quantified relative to 18S rRNA (mean arbitrary units ± SEM). WcRNA from adult and Day 90 fetal bovine liver were used as controls. Adult liver contained 9-fold less IGF-II mRNA than liver from Day 90 fetuses (P < 0.05). Fetal livers of males originating from IVPS and IVPSR groups possessed approximately 2-fold greater levels of mRNA for IGF-II than those from MO males (0.25 ± 0.07, 0.33 ± 0.04, and 0.14 ± 0.03, respectively; P < 0.05). Levels of mRNA for IGF-II tended to be lower (P = 0.07) in skeletal muscle of fetuses originating from the IVPSR group (0.043 ± 0.005) compared to MO controls (0.070 ± 0.008). In conclusion, at Day 70 of gestation, fetuses originating from in vitro production systems possessed altered levels of IGF-II mRNA in both liver and skeletal muscle.}, number={2}, journal={BIOLOGY OF REPRODUCTION}, author={Blondin, P and Farin, PW and Crosier, AE and Alexander, JE and Farin, CE}, year={2000}, month={Feb}, pages={384–389} }
 @article{whaley_hedgpeth_farin_martus_jayes_britt_2000, title={Influence of vitamin A injection before mating on oocyte development, follicular hormones, and ovulation in gilts fed high-energy diets}, volume={78}, DOI={10.2527/2000.7861598x}, abstractNote={Previous research revealed that treatment with vitamin A approximately 5 d before ovulation may increase litter size in weaned sows and improve embryonal survival in gilts fed high-energy diets that reduced embryonal survival. For the current study, the hypothesis was that administration of vitamin A before ovulation would alter development of follicles and oocytes in a way favorable to enhanced embryonal survival. (Landrace x Large White) x (Duroc x Hampshire) gilts (n = 44) were fed 11.0 Mcal ME x gilt(-1) x d(-1) beginning 7 d after second estrus and given (i.m.) corn oil or 1 x 10(6) IU of vitamin A (retinyl palmitate) on d 15 after second estrus. Gilts were checked for estrus every 4 h, mated naturally at third estrus, and assigned randomly to undergo midventral laparotomy beginning at 24 to 28, 28 to 32, 32 to 36, or 36 to 40 h after onset of third estrus. At laparotomy, ovulated oocytes and early-stage embryos were recovered from oviducts, and ovaries were removed for aspiration of oocytes and granulosa cells from unovulated follicles. Oocytes and embryos were stained for assessment of stage of development. Granulosa cells were cultured to assess their ability to secrete progesterone. Follicular fluid was assayed for progesterone, estradiol-17beta, IGF-I, and PGF2alpha. Treatment with vitamin A altered development of oocytes and embryos by decreasing the percentage at the germinal vesicle stage and increasing the percentage at advanced stages. Mean stage of development was increased by vitamin A, but variation in stage was decreased. Among follicles matched by meiotic stage of oocyte, follicular fluid concentrations of progesterone, IGF-I, and PGF2alpha were greater in vitamin A-treated gilts than in controls, but treatment with vitamin A in vivo did not affect LH-stimulated or unstimulated secretion of progesterone by granulosa cells in vitro. These data provide evidence that vitamin A may influence embryonic development by advancing resumption of meiosis and altering follicular hormonal environment during follicle maturation.}, number={6}, journal={Journal of Animal Science}, author={Whaley, S. L. and Hedgpeth, V. S. and Farin, C. E. and Martus, N. S. and Jayes, F. C. L. and Britt, Jack}, year={2000}, pages={1598–1607} }
 @article{ge_nicholson_plotner_farin_gadsby_2000, title={Insulin-like growth factor I receptor Mrna and protein expression in pig corpora lutea}, volume={120}, DOI={10.1530/jrf.0.1200109}, abstractNote={Insulin-like growth factor I (IGF-I) is believed to play a luteotrophic role in the pig corpus luteum during the oestrous cycle. Since the actions of IGF-I in target tissues are mediated by the type I IGF receptor, the concentrations of IGF-I receptor mRNA and protein were examined in pig corpora lutea at different stages of the oestrous cycle. Corpora lutea were collected from normally cyclic gilts on days 4, 7, 10, 13, 15 and 16 of the oestrous cycle (n = 4 animals per day). Corpora lutea on days 7, 10 and 13 were dissociated with collagenase, and large and small luteal cell sub-populations were separated by elutriation. Northern and slot blots were used to examine mRNA, and western blots were used to measure the concentrations of IGF-I receptor protein in the pig corpus luteum. On northern blots, luteal IGF-I receptor mRNA was present as a single 11 kb transcript. The slot blots showed that the steady state expression of IGF-I receptor mRNA increased significantly (P < 0.05) from its lowest value on day 4, to reach a maximum on days 13-16. IGF-I receptor mRNA was also expressed to a greater extent in large compared with small luteal cells (P < 0.05). On western blots, IGF-I receptor appeared as a 95 kDa protein band (beta-subunit) and IGF-I receptor protein concentrations were significantly higher (P < 0.05) on days 4-10 than on days 13-16. Finally, large luteal cells appeared to contain more IGF-I receptor protein than the small luteal cells. In conclusion, since IGF-I receptor was detected in the pig corpus luteum, it is a likely target tissue for IGF-I, especially during the early luteal phase. Furthermore, IGF-I receptor was localized primarily on large luteal cells, thus it is hypothesized that IGF-I may play a paracrine role in the pig corpus luteum.}, number={1}, journal={Journal of Reproduction & Fertility}, author={Ge, Z. and Nicholson, W. E. and Plotner, D. M. and Farin, C. E. and Gadsby, John}, year={2000}, pages={109–114} }
 @article{crosier_farin_dykstra_alexander_farin_2000, title={Ultrastructural morphometry of bovine compact morulae produced in vivo or in vitro}, volume={62}, ISSN={["1529-7268"]}, DOI={10.1095/biolreprod62.5.1459}, abstractNote={Abstract The objective of this study was to compare the ultrastructure of bovine compact morulae produced in vivo or in vitro using morphometric analysis. Compact morulae produced in vivo were obtained from superovulated Holstein cows. Compact morulae produced in vitro were obtained from cumulus-oocyte complexes aspirated from ovaries of Holstein cows. The complexes were matured and fertilized in vitro. At 20 h postinsemination (hpi), zygotes were distributed into 1 of 3 culture media: 1) IVPS (in vitro produced with serum): TCM-199 + 10% estrous cow serum (ECS); 2) IVPSR (in vitro produced with serum restriction): TCM-199 + 1% BSA until 72 hpi followed by TCM-199 + 10% ECS from 72 to 144 hpi; 3) mSOF (modified synthetic oviductal fluid): SOF + 0.6% BSA. At 144 hpi, five grade 1 compact morulae from each of the four treatments were prepared for transmission electron microscopy. The volume density occupied by cellular components was determined by the point-count method using a sampling of seven to nine random micrographs from each compact morula. The volume density of lipid was greater (P < 0.05) in compact morulae from IVPS, IVPSR, and mSOF treatments compared with those produced in vivo. There was a reduced proportional volume of total mitochondria in compact morulae from the IVPS treatment compared with those produced in vivo (P < 0.05). For compact morulae from the IVPS culture treatment, the volume density of vacuoles was greater than that for compact morulae produced in vivo (P < 0.05). The cytoplasmic-to-nuclear ratio for compact morulae from the IVPS treatment was increased (P < 0.05) compared with the ratio for those produced in vivo. In conclusion, compact morulae produced in vitro differed ultrastructurally from those produced in vivo. Compact morulae produced in IVPS culture medium possessed the greatest deviations in cellular ultrastructure.}, number={5}, journal={BIOLOGY OF REPRODUCTION}, author={Crosier, AE and Farin, PW and Dykstra, MJ and Alexander, JE and Farin, CE}, year={2000}, month={May}, pages={1459–1465} }
 @article{blondin_farin_crosier_alexander_farin_1999, title={Does in vitro culture affect the expression of insulin-like growth factor-II (IGF-II) messenger RNA in fetal bovine liver?}, volume={60}, number={1999}, journal={Biology of Reproduction}, author={Blondin, P. and Farin, P. W. and Crosier, A. E. and Alexander, J. E. and Farin, C. E.}, year={1999}, pages={248–249} }
 @article{borchert_farin_washburn_1999, title={Effect of estrus synchronization with norgestomet on the integrity of oocytes from persistent follicles in beef cattle}, volume={77}, DOI={10.2527/1999.77102742x}, abstractNote={Our objective was to determine whether oocyte integrity is compromised when oocytes are recovered from progestogen-induced persistent follicles. Beef cows were presynchronized using PGF2alpha (PGF). Cows detected in estrus after PGF were assigned to either NOR (one 6-mg norgestomet implant for 10 d starting on d 16 of cycle; day 0 = estrus; n = 112) or CON (control, no implant [n = 128] and presynchronized 8 d later than NOR). All cows received 25 mg of PGF at the end of treatment (NOR, d 26; CON, d 18). Treatments produced persistent preovulatory follicles (NOR) or normal preovulatory-size follicles (CON), which were measured via ultrasonography 1 d before slaughter. Ovaries were collected from all animals (NOR, d 27; CON, d 19) along with random (RAN) ovaries from cattle slaughtered on the same days. Cumulus oocyte complexes (COC) were aspirated from the preovulatory follicles with recovery rates of 63% across treatments. Small follicles (2 to 7 mm diameter) from NOR, CON, and RAN cows were also aspirated to recover COC. Preovulatory follicles were larger (19.5+/-.9 vs. 13.6+/-.4 mm, P<.05), serum P4 was lower (.4+/-.1 vs. 3.9+/-.2 ng/mL, P<.05), and serum E2 was higher (28.7+/-1.6 vs. 7.6+/-.8 pg/mL, P<.05) in NOR than in CON cows. Cumulus oocyte complexes recovered from preovulatory follicles (62 NOR, 64 CON) were matured, fertilized, and cultured in vitro for comparison of embryonic development. A subset (24 NOR, 34 CON) of COC were assigned morphological quality grades. A separate set of recovered COC (10 NOR, 15 CON) was fixed within 1 h after recovery for assessment of the stage of meiosis. Treatments did not differ for oocyte quality grade or stage of meiosis. However, COC from NOR cows had more layers of cumulus cells (P<.05), and more of those COC had undergone cumulus expansion (29.2 vs. 5.9%, P<.05 for NOR vs. CON, respectively). Development of cleaved embryos to the morula and blastocyst stages from preovulatory follicles (22.6% NOR, 18.9% CON) or small follicles (42% NOR, 40% CON, 42% RAN) did not differ with treatment. Oocyte quality and in vitro developmental competence were not compromised for oocytes from induced persistent follicles compared with oocytes from normal preovulatory follicles. Increased expansion of cumulus cells associated with oocytes from progestogen-induced persistent follicles may be relevant to the reduction of in vivo fertility associated with such follicles.}, number={10}, journal={Journal of Animal Science}, author={Borchert, K. M. and Farin, C. E. and Washburn, S. P.}, year={1999}, pages={2742–2748} }
 @article{rodriguez_petters_farin_1999, title={Gonadotropin-induced germinal vesicle breakdown in the mouse requires gene transcription.}, volume={60}, number={1999}, journal={Biology of Reproduction}, author={Rodriguez, K. F. and Petters, R. M. and Farin, C. E.}, year={1999}, pages={214} }
 @article{nicholson_ge_plotner_farin_gadsby_1999, title={Insulin-like growth factor (IGF)-I, ICF-I receptor, and IGF binding protein-3 messenger ribonucleic acids and protein in corpora lutea from prostaglandin F-2 alpha-treated gilts}, volume={61}, ISSN={["1529-7268"]}, DOI={10.1095/biolreprod61.6.1527}, abstractNote={Insulin-like growth factor-I (IGF-I) is produced within the porcine corpus luteum (CL) and is thought to play an autocrine/paracrine role in CL development/function during the early luteal phase. This study examines the hypotheses that the luteolytic actions of prostaglandin F(2alpha) (PGF(2alpha)) during the early luteal phase may involve either a decrease in IGF-I or IGF receptor (IGF-IR), or an increase in IGF binding protein (IGFBP)-3, expression, any of which could interfere with the luteotropic actions of IGF-I in this tissue. Cycling gilts were treated twice daily with PGF(2alpha) (or saline) on Days 5-9 of the cycle to induce premature luteolysis. CL were collected on Days 6-9, and RNA, protein, or progesterone was extracted. By slot blot analysis, steady-state levels of IGF-I and IGFBP-3 mRNA were not different in PGF(2alpha)-treated vs. control animals; however, IGF-IR mRNA was increased in treated animals on Day 9. No changes in IGF-I content (ng/CL measured by RIA) were observed with respect to treatment. According to ligand blot analysis, the levels of IGFBP-3 increased on Day 6 and decreased on Days 8-9, while IGFBP-2 was higher on Days 6-7 and decreased on Day 9 in treated animals. IGF-IR levels, determined from Western blots, were higher on Day 7 (P < 0.05) and lower on Day 9 in PGF(2alpha)-treated animals vs. control animals (P < 0.05). In conclusion, PGF(2alpha)-induced premature luteolysis was associated with an increase in steady-state levels of IGF-IR mRNA, but it did not appear to be linked to changes in mRNA levels for IGF-I or IGFBP-3. However, since IGFBP-2 and -3 protein levels increased early in the treatment period (Days 6-7), it is possible that they may mediate the luteolytic actions of PGF(2alpha) by sequestering IGF-I and preventing its interaction with the IGF-IR.}, number={6}, journal={BIOLOGY OF REPRODUCTION}, author={Nicholson, WCE and Ge, ZP and Plotner, DM and Farin, CE and Gadsby, JE}, year={1999}, month={Dec}, pages={1527–1534} }
 @article{farin_hasler_martus_stokes_1997, title={A comparison of Menezo's B2 and tissue culture medium-199 for in vitro production of bovine blastocysts}, volume={48}, ISSN={["0093-691X"]}, DOI={10.1016/S0093-691X(97)00294-X}, abstractNote={The objectives of this study were, first, to evaluate the effectiveness of 2 culture media, Menezo's B2 (B2) and Tissue Culture Medium-199 (M-199), for the production of bovine blastocysts in a commercial embryo transfer program; and, second, to characterize the stage of development, quality grade and cell number of blastocysts produced in each medium. One-cell bovine embryos were produced using in vitro maturation and fertilization procedures. After fertilization, the embryos were co-cultured on Buffalo rat liver (BRL) cell monolayers in either B2 or M-199+1% BSA (M-199) medium. Both media were supplemented with 10% fetal calf serum (FCS) and penicillin/streptomycin. Embryo cultures were continued undisturbed to either Day 7 or Day 8 post-insemination. In the Day 7 cultures, all blastocysts were removed for evaluation on Day 7, and the remaining embryos were cultured for a further 24 h. Any additional blastocysts that formed were removed for evaluation and designated as Day 8 disturbed embryos. All blastocysts were classified for stage and quality grade. Embryos were fixed and stained for determination of cell number. Overall, the proportion of blastocysts was greater (P = 0.0003) with B2 medium (46%) than with M-199 (33%). This was due to a larger (P = 0.0001) proportion of blastocysts produced in B2 medium when cultures were left undisturbed for 8 d (50 vs 28% for B2 vs M-199). The proportion of blastocysts on Day 7 of culture tended to differ (P = 0.073) between media (33 vs 24% for B2 vs M-199). In addition, there were more (P = 0.007) blastocysts at advanced stages of development in B2 medium on Day 7. There was no effect of type of medium on the distribution of embryo quality grades on any day examined. The number of cells per blastocyst did not differ between media but did vary significantly (P < .05) with both stage and grade. In conclusion, B2 medium was superior to M-199 medium when used in a co-culture system with BRL cells for the production of bovine blastocysts.}, number={5}, journal={THERIOGENOLOGY}, author={Farin, CE and Hasler, JF and Martus, NS and Stokes, JE}, year={1997}, month={Oct}, pages={699–709} }
 @article{piscopo_farin_bai_1997, title={Determination of concentration of albendazole sulfoxide in plasma and uterine fluid of heifers}, volume={58}, number={1}, journal={American Journal of Veterinary Research}, author={Piscopo, S. E. and Farin, C. E. and Bai, S. A.}, year={1997}, pages={62–65} }
 @article{farin_yang_1994, title={INHIBITION OF GERMINAL VESICLE BREAKDOWN IN BOVINE OOCYTES BY 5,6-DICHLORO-1-BETA-D-RIBOFURANOSYLBENZIMIDAZOLE (DRB)}, volume={37}, ISSN={["1098-2795"]}, DOI={10.1002/mrd.1080370307}, abstractNote={Abstract}, number={3}, journal={MOLECULAR REPRODUCTION AND DEVELOPMENT}, author={FARIN, CE and YANG, L}, year={1994}, month={Mar}, pages={284–292} }