@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{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{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{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{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{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} }