@article{harlow_griesgraber_seman_shuping_sommer_griffith_hileman_nestor_2022, title={The impact of undernutrition on KNDy (kisspeptin/neurokinin B/dynorphin) neurons in female lambs}, volume={5}, ISSN={["1365-2826"]}, url={https://doi.org/10.1111/jne.13135}, DOI={10.1111/jne.13135}, abstractNote={AbstractUndernutrition limits reproduction through inhibition of gonadotropin‐releasing hormone (GnRH)/luteinizing hormone (LH) secretion. Because KNDy neurons coexpress neuropeptides that play stimulatory (kisspeptin and neurokinin B [NKB]) and inhibitory (dynorphin) roles in pulsatile GnRH/LH release, we hypothesized that undernutrition would inhibit kisspeptin and NKB expression at the same time as increasing dynorphin expression. Fifteen ovariectomized lambs were either fed to maintain pre‐study body weight (controls) or feed‐restricted to lose 20% of pre‐study body weight (FR) over 13 weeks. Blood samples were collected and plasma from weeks 0 and 13 were assessed for LH by radioimmunoassay. At week 13, animals were killed, and brain tissue was processed for assessment of KNDy peptide mRNA or protein expression. Mean LH and LH pulse amplitude were lower in FR lambs compared to controls. We observed lower mRNA abundance for kisspeptin within KNDy neurons of FR lambs compared to controls with no significant change in mRNA for NKB or dynorphin. We also observed that FR lambs had fewer numbers of arcuate nucleus kisspeptin and NKB perikarya compared to controls. These findings support the idea that KNDy neurons are important for regulating reproduction during undernutrition in female sheep.}, number={6}, journal={JOURNAL OF NEUROENDOCRINOLOGY}, publisher={Wiley}, author={Harlow, KaLynn and Griesgraber, Max J. and Seman, Andrew D. and Shuping, Sydney L. and Sommer, Jeffrey R. and Griffith, Emily H. and Hileman, Stanley M. and Nestor, Casey C.}, year={2022}, month={May} }
 @article{harlow_renwick_shuping_sommer_lents_knauer_nestor_2021, title={Evidence that pubertal status impacts kisspeptin/neurokinin B/dynorphin neurons in the gilt(dagger)}, volume={105}, ISSN={["1529-7268"]}, DOI={10.1093/biolre/ioab189}, abstractNote={Abstract
               Puberty onset is a complex physiological process, which enables the capacity for reproduction through increased gonadotropin-releasing hormone and subsequently luteinizing hormone secretion. While cells that coexpress kisspeptin, neurokinin B (NKB), and dynorphin in the hypothalamic arcuate nucleus are believed to govern the timing of puberty, the degree to which kisspeptin/NKB/dynorphin (KNDy) neurons exist and are regulated by pubertal status remains to be determined in the gilt. Hypothalamic tissue from prepubertal and postpubertal, early follicular phase gilts was used to determine the expression of kisspeptin, NKB, and dynorphin within the arcuate nucleus. Fluorescent in situ hybridization revealed that the majority (>74%) of arcuate nucleus neurons that express mRNA for kisspeptin coexpressed mRNA for NKB and dynorphin. There were fewer arcuate nucleus cells that expressed mRNA for dynorphin in postpubertal gilts compared to prepubertal gilts (P < 0.05), but the number of arcuate nucleus cells expressing mRNA for kisspeptin or NKB was not different between groups. Within KNDy neurons, mRNA abundance for kisspeptin, NKB, and dynorphin of postpubertal gilts was the same as, less than, and greater than, respectively, prepubertal gilts. Immunostaining for kisspeptin did not differ between prepubertal and postpubertal gilts, but there were fewer NKB immunoreactive fibers in postpubertal gilts compared to prepubertal gilts (P < 0.05). Together, these data reveal novel information about KNDy neurons in gilts and support the idea that NKB and dynorphin play a role in puberty onset in the female pig.}, number={6}, journal={BIOLOGY OF REPRODUCTION}, author={Harlow, KaLynn and Renwick, Allison N. and Shuping, Sydney L. and Sommer, Jeffrey R. and Lents, Clay A. and Knauer, Mark T. and Nestor, Casey C.}, year={2021}, month={Dec}, pages={1533–1544} }
 @article{aerts_harlow_griesgraber_bowdridge_hardy_nestor_hileman_2021, title={Kisspeptin, Neurokinin B, and Dynorphin Expression during Pubertal Development in Female Sheep}, volume={10}, ISSN={["2079-7737"]}, url={https://doi.org/10.3390/biology10100988}, DOI={10.3390/biology10100988}, abstractNote={The neural mechanisms underlying increases in gonadotropin-releasing hormone (GnRH) and luteinizing hormone (LH) secretion that drive puberty onset are unknown. Neurons coexpressing kisspeptin, neurokinin B (NKB), and dynorphin, i.e., KNDy neurons, are important as kisspeptin and NKB are stimulatory, and dynorphin inhibitory, to GnRH secretion. Given this, we hypothesized that kisspeptin and NKB expression would increase, but that dynorphin expression would decrease, with puberty. We collected blood and hypothalamic tissue from ovariectomized lambs implanted with estradiol at five, six, seven, eight (puberty), and ten months of age. Mean LH values and LH pulse frequency were the lowest at five to seven months, intermediate at eight months, and highest at ten months. Kisspeptin and NKB immunopositive cell numbers did not change with age. Numbers of cells expressing mRNA for kisspeptin, NKB, or dynorphin were similar at five, eight, and ten months of age. Age did not affect mRNA expression per cell for kisspeptin or NKB, but dynorphin mRNA expression per cell was elevated at ten months versus five months. Thus, neither KNDy protein nor mRNA expression changed in a predictable manner during pubertal development. These data raise the possibility that KNDy neurons, while critical, may await other inputs for the initiation of puberty.}, number={10}, journal={BIOLOGY-BASEL}, author={Aerts, Eliana G. and Harlow, KaLynn and Griesgraber, Max J. and Bowdridge, Elizabeth C. and Hardy, Steven L. and Nestor, Casey C. and Hileman, Stanley M.}, year={2021}, month={Oct} }
 @article{harlow_renwick_shuping_sommer_knauer_nestor_2020, title={Effects of genetic selection for early puberty on the hypothalamic-pituitary-ovarian axis in gilts}, volume={98}, ISSN={["1525-3163"]}, DOI={10.1093/jas/skaa054.368}, abstractNote={Abstract
               Puberty onset in gilts is an awakening of the hypothalamic-pituitary-ovarian axis that is the result of reduced estradiol-negative feedback at the level of the hypothalamus which yields elevated gonadotropin secretion from the anterior pituitary. Given the importance of hypothalamic kisspeptin and neurokinin B (NKB) signaling for the onset of puberty in other species, the objective of this study was to determine if gilts selected for early pubertal onset (SELECT) would display measurable differences within the hypothalamus (i.e. increased expression of kisspeptin and NKB) and within the ovary (i.e. increased ovarian mass) compared to age-matched and weight-matched gilts (CONTROL) that achieve puberty 20 days later than SELECT gilts. Gilts were sacrificed at three timepoints: Timepoint A, both groups were determined to be prepubertal (n=6/group), Timepoint B, SELECT gilts were determined to be pubertal and CONTROL gilts were determined to be prepubertal (n=6/group), and Timepoint C, both groups were determined to be pubertal (n=6/group). All animals were euthanized, heads were perfused with 8 L of 4% paraformaldehyde, and ovaries were harvested. Brain tissue was removed post-fixation, submerged in fixative for 24 hrs followed by 20% sucrose until sectioned for immunohistochemistry. Ovarian mass tended (p≤0.10) to be greater for SELECT gilts on the right ovary (4.34 vs. 3.67 g) and the left ovary (4.49 vs. 3.68 g) when compared to CONTROL (Timepoints A and C), and at Timepoint B right ovary mass from SELECT gilts was heavier than CONTROL gilts (p< 0.05; 7.22 vs. 4.65 g). Hypothalamic immunohistochemistry for kisspeptin and NKB revealed differences in neuronal fiber density between both groups at various timepoints. Therefore, we conclude that gilts genetically selected for early puberty do so via changes within the hypothalamus that increase gonadotropin secretion and, in turn, stimulate ovarian growth to ultimately advance the timing of puberty onset.}, journal={JOURNAL OF ANIMAL SCIENCE}, author={Harlow, KaLynn and Renwick, Allison and Shuping, Sydney and Sommer, Jeff and Knauer, Mark and Nestor, Casey}, year={2020}, month={Nov}, pages={212–212} }
 @article{harlow_ferreira_sobreira_casey_stewart_2019, title={Lipidome profiles of postnatal day 2 vaginal swabs reflect fat composition of gilt’s postnatal diet}, volume={3}, url={https://doi.org/10.1101/593392}, DOI={10.1101/593392}, abstractNote={AbstractWe hypothesized that postnatal development of the vagina is impacted by early nutritional environment. Our objective was to determine if lipid profiles of vaginal swabs were different between gilts suckled by sow or fed milk replacer the first 48 h postpartum, with and without a lard-based fat supplement. Gilts (>1.3 kg) were selected at birth across 8 litters and assigned to treatments: colostrum suckled (S, n=8); S plus fat supplement (SF, n=5); bottle-fed milk replacer (B, n=8); or B plus fat supplement (BF, n=7). At 48 h postnatal, vaginal swabs were taken with a cytology brush, immersed in ultrapure water to burst cells, and lipids extracted for analysis using multiple reaction monitoring (MRM)-profiling. Lipids extracted from serum collected at 48 h from gilts and milk collected from sows at 24 h were also analyzed with MRM-profiling. Receiver operating characteristic curve analysis found 18 lipids highly distinguished [area-under-the-curve (AUC) > 0.9] between S and B gilts, including phosphatidylethanolamine with 34 carbon and four unsaturations in the fatty acyl residues [PE(34:4)]. Twelve lipids from vaginal swabs highly correlated (r> 0.6;p< 0.01) with nutrition source. Lipids more abundant in milk replacer drove association. For example, mean intensity of PE (34:4) was 149-fold higher in milk replacer than colostrum, with 1.6- and 2.12-fold higher levels in serum and vaginal swab samples (p< 0.001), respectively, of B versus S gilts. Findings support that vaginal swabs can be used to noninvasively study effects of perinatal nutrition on tissue composition.Summary sentenceVaginal swab lipidome profiles at 48 h reflect the fat composition of neonatal diet during first two days postnatal.}, publisher={Cold Spring Harbor Laboratory}, author={Harlow, KaLynn and Ferreira, Christina R. and Sobreira, Tiago J.P. and Casey, Theresa and Stewart, Kara}, year={2019}, month={Mar} }
 @article{harlow_ferreira_sobreira_casey_stewart_2019, title={Lipidome profiles of postnatal day 2 vaginal swabs reflect fat composition of gilt’s postnatal diet}, url={https://doi.org/10.1371/journal.pone.0215186}, DOI={10.1371/journal.pone.0215186}, abstractNote={We hypothesized that postnatal development of the vagina is impacted by early nutritional environment. Our objective was to determine if lipid profiles of vaginal swabs were different between gilts suckled by sow or fed milk replacer the first 48 h postpartum, with and without a lard-based fat supplement. Gilts (>1.3 kg) were selected at birth across 8 litters and assigned to treatments: colostrum suckled (S, n=8); S plus fat supplement (SF, n=5); bottle-fed milk replacer (B, n=8); or B plus fat supplement (BF, n=7). At 48 h postnatal, vaginal swabs were taken with a cytology brush, immersed in ultrapure water to burst cells, and lipids extracted for analysis using multiple reaction monitoring (MRM)-profiling. Lipids extracted from serum collected at 48 h from gilts and milk collected from sows at 24 h were also analyzed with MRM-profiling. Receiver operating characteristic curve analysis found 18 lipids highly distinguished [area-under-the-curve (AUC) > 0.9] between S and B gilts, including phosphatidylethanolamine with 34 carbon and four unsaturations in the fatty acyl residues [PE(34:4)]. Twelve lipids from vaginal swabs highly correlated (r > 0.6; p < 0.01) with nutrition source. Lipids more abundant in milk replacer drove association. For example, mean intensity of PE (34:4) was 149-fold higher in milk replacer than colostrum, with 1.6- and 2.12-fold higher levels in serum and vaginal swab samples (p < 0.001), respectively, of B versus S gilts. Findings support that vaginal swabs can be used to noninvasively study effects of perinatal nutrition on tissue composition. Summary sentence Vaginal swab lipidome profiles at 48 h reflect the fat composition of neonatal diet during first two days postnatal.}, journal={PLOS ONE}, author={Harlow, KaLynn and Ferreira, Christina R. and Sobreira, Tiago J. P. and Casey, Theresa and Stewart, Kara}, editor={Loor, Juan J.Editor}, year={2019}, month={Sep} }
 @article{temporal analysis of vaginal proteome reveals developmental changes in lower reproductive tract of gilts across the first two weeks postnatal_2019, url={http://dx.doi.org/10.1038/s41598-019-49597-w}, DOI={10.1038/s41598-019-49597-w}, abstractNote={AbstractIn swine the upper reproductive tract undergoes early postnatal development, however little is known about the lower reproductive tract. Our objective was to measure cytology and proteome of vaginal swab samples taken on postnatal day (PND) 2 and 16 in gilts to determine if temporal changes occurred in cell and protein profiles during the first two weeks after birth. The posterior vagina was swabbed using a cytology brush on PND 0, 2 and 16 and slides were prepared. The proportion of anuclear and superficial cells increased and parabasal decreased (P < 0.05) from PND 0 to 16. Proteins isolated from vaginal swabs taken on PND 2 and 16 from six gilts across three litters were measured using LC-MS/MS. Over 1500 proteins were identified, with 881 differentially expressed (P-adj < 0.05) between PND 2 and 16. One-third of proteins upregulated between days were categorized as secreted, including lipocalins. Categories enriched by downregulated proteins included cell-cell adherens junction, translation and ER to Golgi vesicle-mediated transport, and reflected increased cornification of stratified epithelium and thus mirrored changes in cytology. Changes in cytology and proteome over the first two weeks after birth support that the porcine vagina continues to develop postnatal.}, journal={Scientific Reports}, year={2019}, month={Dec} }
 @article{diet impacts pre-implantation histotroph proteomes in beef cattle_2018, url={http://dx.doi.org/10.1021/acs.jproteome.8b00077}, DOI={10.1021/acs.jproteome.8b00077}, abstractNote={In ruminants, the period from fertilization to implantation is relatively prolonged, and the survival of embryos depends on uterine secretions known as histotroph. Our objective was to determine if the pre-breeding diet affected histotroph proteomes in beef cattle. Cows were assigned to one of four diets: a control diet (CON), a high-protein diet (PROT), a high-fat diet (OIL), or a high-protein and high-fat diet (PROT + OIL). After 185 days on these diets, an intravaginal progesterone implant (CIDR) was inserted for 7 days. At 9 days after CIDR removal, animals with a corpus luteum were selected ( n = 16; 4 per treatment). Proteins were isolated from the histotroph collected by uterine lavage and analyzed with liquid chromatography-tandem mass spectrometry. Over 2000 proteins were expressed ( n ≥ 3 cows per treatment), with 1239 proteins being common among all of the groups. There were 20, 37, 85, and 123 proteins unique to CON, PROT + OIL, PROT, and OIL, respectively. Relative to CON, 23, 14, and 51 proteins were differentially expressed in PROT + OIL, PROT, and OIL, respectively. Functional analysis found that 53% of histotroph proteins were categorized as extracellular exosome, 3.28% as cell-cell adhesion, and 17.4% in KEGG metabolic pathways. Differences in proteomes among treatments support the idea that pre-breeding diet affects histotroph. Understanding the impact of diet on histotroph proteins may help improve conception rates.}, journal={Journal of Proteome Research}, year={2018}, month={Jun} }
 @article{the potential of identifying replacement gilts by screening for lipid biomarkers in reproductive tract swabs taken at weaning_2018, url={http://dx.doi.org/10.1080/09712119.2017.1384733}, DOI={10.1080/09712119.2017.1384733}, abstractNote={Sow longevity affects economic returns to pork producers. The cost of gilt replacements is substantial and sows with greater than three litters have lower costs per pig produced. An early marker of reproductive potential would facilitate early identification of superior females, and likely increase sow longevity. Gilts raised in small litters have greater reproductive competence, but mechanisms associated with increased reproductive responses are not fully understood. Here, early postnatal development of the gilt's reproductive tract is described, and a brief review of literature is presented to support that factors in colostrum regulate the developmental trajectory of the gilt's uterine tissues. We propose that, similar to the uterus, nutritional environment likely affects the postnatal developmental programme of the vagina. A metabolomics approach, multiple reaction monitoring -profiling, for biomarker discovery is described, along with evidence that lipids present in vaginal samples are differentially expressed in gilts exposed to colostrum versus milk replacer fed. These exploratory studies indicate that the vaginal cell lipidome may reflect the postnatal nutritional environment, which defines to a large extent the gilt's reproductive potential. Together findings support further investigations to identify biomarkers predictive of fertility outcomes in the metabolome of gilt reproductive tracts.}, journal={Journal of Applied Animal Research}, year={2018}, month={Jan} }