Brief Summary of Minutes of Annual Meeting

Chair Mike Day called the meeting to order at 8:30 am on Monday, May 17, 2010. Minutes from the 2009 meeting were approved without discussion. Tom Adams was thanked for making arrangements for this year's meeting. Colin Kaltenbach requested an update of the membership list for NIMSS files and noted that that the project terminates in approximately 15 months. A renewal of the project is due by January 15, 2011 and needs to be submitted to the Western Directors in March, 2011. To meet those deadlines, the re-write of the project must be completed by December 15, 2010. Dennis Hallford, Tracy Davis, Mike Teglas, and John Stevens agreed to develop new objectives for presentation to the group on Tuesday morning. Adelle Turzillo provided a handout regarding new developments at NIFA. We were encouraged to submit stakeholder comments about AFRI programs and to review AFRI-RFAs carefully. Four of the seven AFRI-RFA programs contain an animal component that may fit with our research programs. The trend for new grants is towards large multi-disciplinary projects rather than multiple small grants. Last year's election of Kim Vonnahme as Member-at-Large for 2009-2010 was confirmed. Tracy Davis was selected as the Member-at-Large for 2010-2011. Hawaii was selected as the site for next year's meeting on May 16-17, 2011. Updates and announcements were made by each station regarding new faculty, positions available and ongoing changes. Registration costs requested for meeting attendance were $60.00. The remainder of the day was dedicated to station reports. The meeting adjourned at 5:00 p.m. for the day. Tuesday May 18, 2010. The meeting was called to order at 8:30 a.m. The first business of the day concerned the re-write of the project for the 2012-2017. New objectives for the re-write were presented, adapted, and approved to read as follows: Objective 1. Examine molecular, metabolic, genomic, endocrine, and immunologic mechanisms influencing testicular and ovarian function, reproductive behavior, conception rate, embryo and fetal development, attainment of puberty, and effects of climate/season on reproductive patterns of domestic ruminants. Objective 2. Development and application of methodologies to improve animal health, well-being, and reproductive efficiency of domestic ruminants in order to allow sustainable operation of production systems in the western United States. Subsequent discussion focused on procedures and formation of a writing committee for the project's renewal. Tom Geary will be solicited by Mike Day to prepare the summary of accomplishments for the past five years of the project. All participants will be asked to submit a paragraph of their research plans for the next five years as they pertain to the newly adopted objectives. These paragraphs will be due to Gary Moss by September 15, 2010. Gary Moss, Mike Teglas, and Terry Nett and nearby participants (esp. at WY and CO) will coordinate the collation of those paragraphs into a readable document for submission by December 15, 2010. The remainder of the morning was dedicated to the completion of station reports and discussion of collaborative research plans. The meeting was adjourned at 12:00. Collaborative Efforts for the coming year: NM/MT. Metabolic hormone patterns in ewe lambs with differing RFIs. NM/CO. Nutrient restriction vs. metabolic hormone patterns NM/CO. Thyroid involvement in BVD infected cows. NM/AR. Metabolic hormone profiles in cows consuming endophyte infected fescue. NM/OH. Follicular fluid IGF. WY/MT/NM. Meta-analysis of birth type on ewe reproductive performance. CO/OH ISG mRNAs in maternal blood and IFNt in conceptus. CO/WA use mouse ISG15 knock out model to understand the role if ISG15 in bovine uterus. CO/HI/TX PGEs and CL function using intraluteal implants AK/WY/MN Influence of progesterone on sexual behavior AZ/CO Influence of gestational nutrition on pancreatic function AZ/WY Influence of gestational nutrition on insulin MI/FL Extended oocyte/embryo culture MI/MT Oocyte quality and follicle diameter OH/MT Follicle age influences on fertility OH/PA MX and INFt in cattle MT/MI Oocyte quality NV/TX Stress hormones and abortion in cattle and sheep TX/MN Inhibin concentrations and temperament WT AM/NM Development in intact males ND/WA Fetal programming WY/MT/NE Progesterone and sexual behavior WY/UT SNPs in sheep AR/NM Pasture systems and reproduction WA/NE Role of VEGF in Spermatogonial Stem cell development and differentiation.

Objective 1. Investigate molecular, cellular and endocrine mechanisms that limit or control reproductive efficiency in domestic animals. Estrous cycle, physiology/CL function: Dimeric inhibin is present in the plasma of ram lambs and yearling ewes. In addition to dimeric inhibin, both ram lambs and yearling ewes appear to have an excess of monomeric inhibin ±-subunit. Attempts to detect dimeric inhibin in ovine plasma using a two-site ELISA format have been unsuccessful. Present findings confirm that dimeric inhibin is indeed present in ovine plasma. Plasma ±-inhibin in ram lambs consists of two ±-subunits, 47 and 17 kDa, and one ²-subunit of 14 kDa. The 47 kDa ±-subunit is most prominent. This larger ±-band is consistent with non-biologically active free inhibin ±-subunit. The smaller 17 kDa ±-band and the 14 kDa ²-band are consistent with biologically active dimeric inhibin. Thus, the ±RIA, which detects both the 47 and 17 kDa forms of the inhibin ±-subunit, does not necessarily reflect biologically active dimeric inhibin. Plasma ±-inhibin in yearling ewes consists of 46 and 14 kDa ±-subunits and a single 14 kDa ²-subunit. The inhibin forms are either very similar or identical to those in ran lambs. The intensity of immunostaining of the 47 kDa ±-band in ram lambs was greater than the 46 kDa ±-band in yearling ewes. Concentrations of ±-inhibin increased decreased immediately following CIDR removal and then increased. Dynamic differences exist in the gene expression profile and accompanying functional gene categories and biological pathways regulated by PG in bovine CL that are responsive versus refractory to the luteolytic actions of PG. Such differences are linked to candidate transcription factors that may potentially mediate the stage specific gene expression response to PG associated with luteolysis. PGE1 or PGE2 prevent luteolysis in Angus or Brahman cows by preventing loss of message for LH receptors and occupied and unoccupied LH receptors and alter mRNA for prostanoid receptors on luteal tissue. Analogues of PGE1 and/or PGE2 may be viable approaches to prevent loss of luteal progesterone in cows and to decrease pregnancy loss during the first trimester of pregnancy. Hypothalamic-Anterior Pituitary Gland-Ovarian Physiology Using a lentiviral-based siRNA knockdown strategy, it was established that PGRMC1 is necessary for mediating the anti-apoptotic actions of progesterone. PGRMC1 deficiency was found to reduce tumor growth and enhance sensitivity to chemotherapy using an immunocompromised mouse xenograft model. Gene expression for proteins regulating folliculogenesis was higher in the Suffolk ewes compared to the Polypay. There were no observed differences in the number of preantral and antral follicles between the two breeds. Recombinant single chain analogs of ovine gonadotropins can be used to induce a superovulatory response in sheep and may have broader application in ovulation induction protocols. Gonadal morphology in VEGF Sertoli and Granulosa cell KO mice (using pDMRT1) is altered and their gonads are smaller which may result in reduction in fertility plus abnormal structures on the ovaries. Potential differences in ovarian reserve in the females and alteration in endocrine profiles in both sexes (high androgen in males and reduced estrogen in females) are being analyzed. Gonadal morphology in VEGF granulose cell KO mice (using AMHR2) is similar to that of VEGFXDMRT1 mice in the ovaries are smaller and have less follicles. The VEGFAMHR2 cre positive mice have reduced body size and have more dramatic reductions in estrogen over cre negative control mice. Steroidogenic enzymes to determine if there are differences that may result in reduced estrogen concentrations in blood plasma are being evaluated. Eliminating a co-receptor for VEGF, neuropillin-1, decreases ovarian size and reduces the number of follicles on the ovary. The data is preliminary, but suggests that angiogenic VEGF isoforms are necessary for appropriate granulosa cell function and follicle development. Oocyte-embryo-uterine physiology: Preovulatory concentrations of estradiol did not influence amount of uterine mRNA for ISG-15 and Mx2 or embryonic mRNA for CSH-1 and IFNt on d 17.5 of gestation/estrous cycle. Glucocorticoid exposure during the second half of pregnancy has deleterious effects on pregnancy outcomes. Our findings over the past year indicate that the embryo signals within the endometrium shift toward local production of glucocorticoids. It is hypothesized that activation of GRs within the endometrium results in establishment of a barrier that precludes access of maternal immune cells to the embryo through increased expression of adhesion molecules. Through the use of conditional knockout mice, it was established that b-catenin signaling in the stromal compartment of the endometrium is essential for coordinating epithelial responses to steroid hormones (i.e., mesenchymal-epithelial interactions). It was established through microarray analysis and validation by RT-PCR and in situ hybridization that the embryo induces endometrial expression of an as yet uncharacterized gene referred to as decidual restricted gene 1. It is hypothesized that Drg1 is functionally required for the establishment and maintenance of pregnancy. It is possible that synchronization of estrus of beef heifers with prostaglandin F2± may impair function of the corpus luteum in the ensuing estrous cycle. This may be of physiological significance relative to early embryo survival. The effect of follicle size on fertility may be manifested primarily through estradiol effects on the uterus since cows induced to ovulate while serum estradiol concentration was low had reduced ability to maintain a pregnancy to d 28. Also, d 28 serum concentration of PAGs may be indicative of pregnancies that will not be supported to term. Differential methylation patterns may help explain large offspring syndrome in calves born to IVM-IVF and cloning procedures and could serve as a model for study of a similar condition in humans. JY-1 induced regulation of cumulus expansion is dependent on presence of additional oocyte- secreted factors and potentially mediated, at least in part, by regulation of PGE2 production and expression of genes linked to stabilization of the hyaluronan rich extracellular matrix. Fibroblast growth factors (FGFs) stimulate IFNT production in bovine trophectoderm. Several FGFs are expressed in bovine endometrium and conceptuses. FGF2 promotes endoderm development in bovine embryos. Also, FGF2 and 10 stimulate trophectoderm migratory activity. Endogenous FGFs are important for normal bovine embryonic development. Blocking FGR activity with specific kinase-domain inhibitors does not affect blastocyst formation in vitro but compromises subsequent development and function. FGFs also promote oocyte competency. Embryo development in vitro is improved by supplementing FGF2 or 10 during in vitro maturation. Also, treatment with FGFR inhibitors during maturation limits subsequent embryo development. A FACS-based approach was developed to obtain enriched TGC samples for study of the events that lead to late embryonic and fetal losses in cattle so that new schemes can be developed to limit the severity of these losses. Current work focused on developing ways to study trophoblast giant cells (TGCs) from bovine placentae. Ovine trophoblast cells exhibit phenotypic changes, when cultured on Matrigel matrix, similar to what is observed by mouse trophoblast stem cells and first trimester human cytotrophblasts, exhibiting migratory and invasive behavior. This may suggest that the phenotype of trophoblast cells may not differ significantly between non-invasive ruminant conceptuses compared to invasive human and mouse conceptuses, inferring that the differences in placentation result from functional differences in the uterus. Culture of ovine trophoblast cells on Matrigel matrix upregulates the expression of PRR15, allowing a more robust system to examine its function and regulation. IFNT has endocrine effects on the CL between Days 13 and 14 of pregnancy and may protect the CL through mechanisms that are complementary, yet independent to its paracrine effects on the OXTR endometrial pathway. CXCL12/CXCR4 pathway is activated during implantation and placentation in sheep and is likely playing a role in the communication between trophoblast cells and the maternal endometrium. When added to culture media, bovine ghrelin significantly reduces bovine embryonic development Testicular Physiology: Effects of VEGF on testis development and spermatogenesis were evaluated to improve our understanding of factors critical for spermatogonial stem cell differentiation and maintenance in cattle. Testis morphology in VEGF Sertoli and Granulosa cell KO mice is altered and their gonads are smaller which may result in reduction in fertility plus the testes appear to have less numbers of sperm. VEGF anti-angiogenic isoforms may increase germ cell apoptosis and may, therefore, negatively affect spermatogenesis and male fertility. VEGF anti-angiogenic isoforms may increase germ cell apoptosis therefore factors that increase amounts of these isoforms may negatively affect spermatogenesis and male fertility. Uterine-Placental-Fetal Nutritional Interactions during Pregnancy The application of a powerful image analyses program has allowed vascular development to be quantified in the sheep placentome, and to determine if morphology of the ovine placentome impacts vascularity. Moreover, impacts of maternal diet on placental vascular function are being assessed. Early- to mid-gestation maternal undernutrition in twin sheep pregnancies, followed by realimentation appears to set the fetus on a more rapid growth trajectory during late gestation, which may be carried forward into the postnatal period. Maternal undernutrition may enhance the pregnancy induced insulin resistance, such that upon realimentation, maternal uterine artery glucose concentrations are significantly elevated, even when compared to the control fed ewes receiving the same diet. While uterine artery, umbilical vein and umbilical artery glucose concentrations do not differ between control and continuously undernourished twin pregnancies, the umbilical vein:umbilical artery difference in glucose concentration is significantly less in continuously undernourished pregnancies. This may suggest that even with normal glucose delivery to the fetus, that fetal metabolism is still reduced. Reductions in metabolic set points within these offspring may well impact postnatal growth rate, feed efficiency and body composition. Models of both maternal undernutrition and maternal obesity have been established to allow evaluation and comparison of the impacts of each condition on fetal growth and development, as well as offspring health, growth efficiency, glucose-insulin dynamics and reproductive function. Results from these studies will shed light on the specific physiological mechanisms involved, as well as lead us to management practices to optimize offspring health and production efficiency. This is especially pertinent when one considers that in the human, both maternal undernutrition and maternal obesity lead to the birth of offspring which exhibit an increased incidence of obesity, insulin resistance, type 2 diabetes and cardiovascular disease in later life. By studying both maternal under and overnutrition simultaneously, we can elucidate any similarities on fetal growth and development as well as on offspring postnatally. Optimizing the uterine environment to facilitate the birth of healthy, growth-efficient offspring is important for livestock production Female offspring born to ewes nutrient restricted (NR) during the first half of gestation exhibited a marked reduction in fertility. This decreased fertility was associated with decreased progesterone concentrations in systemic blood during the estrous cycle of adult ewes born to NR mothers. Corpora lutea recovered from the adult female offspring of these NR ewes exhibited markedly decreased progesterone content during the midluteal phase of an estrous cycle. CLs are currently being evaluated both histologically and for selected mRNA and protein expression differences to determine the reason for this difference. Chronically elevated norepinephrine, without hypoxemia, continually suppresses plasma insulin concentrations in fetal sheep. Termination of the norepinephrine infusion results in hyper-insulin secretion in response to glucose and arginine, showing chronic suppression by NE causes ²-cell compensation. Insulin secretion from MIN6 cells is suppressed by the adrenergic receptor ±2, similar to rat and mouse islets. Therefore, MIN6 cells provide a useful culture system to examine alterations in insulin secretion responsiveness following chronic catecholamine suppression. In IUGR fetuses, islet vascularity is reduced to a greater extent than ²-cell area. These findings begin to link disruptions in endothelial cell function and islet blood flow as mechanism for decrease islet function in utero. Statistical Methodology: Additional statistical models (and preprocessing methods) were assessed using spike-in data, specifically FIRSTP (Rubin 2009) and RMANOVA (Xu and Cui 2008), under both RMA and GCRMA. An appropriate sampling distribution of test statistics from the nested factorial model was determined, using permutation methods. This allows computation of traditionally reported P-values for each gene. The developed NFM method was found to be competitive against other statistical models, and was applied to the motivating bovine NT data (4 clone vs. 3 control arrays, in cotyledon). The resulting 584 significant genes were found to be biologically relevant. The developed NFM method was implemented in R code (affyNFM), now freely available on the project leaders website (www.stat.usu.edu/~jrstevens). Nutrition and Reproduction: Steroidogenic capacity of dominant follicles was not influenced consistently by diets designed to induce precocious puberty. Studies continue to determine how selenium being fed impacts maternal and fetal tissues during pregnancy. Data indicate maternal nutrition impacts circulating hormones, as well as maternal and fetal placental tissues from the offspring. Further data are being analyzed for growth parameters and carcass composition of the offspring born from dams fed supranutritional levels of selenium as well as differing global levels of nutrition. Both over- and under-nutrition impact colostrum yield and IgG production. Milk production in ewes that are underfed continue to be lower than control fed ewes. Ewes that are overfed will have decreased colostrum, but will have similar milk production 3 to 4 days after parturition. Level of harvested feed provided to cows during the winter appears to impose a uterine programming influence that alters circulating concentrations of IGF-1 and reproductive performance in their daughters when they are subsequently reared on restricted levels of nutritional intake. Heifers developed on restricted diets from dams wintered on marginal levels of feed had greater concentrations of IGF-1 from precalving through 2nd prebreeding and greater retention rates to their 5th breeding season than contemporary heifers that were out of dams provided adequate supplemental forage throughout the winter. Residual feed intake did not influence resumption of luteal function during the transition into the breeding season or temporal patterns of progesterone and metabolic hormones. Ewes classified as efficient RFI gained more weight during ram exposure than ewes classified as efficient and not exposed to wethers. Behavior and Stress: Research related to objective identification of temperament, and the coupling of stress-responsiveness with immune function, provide added impetus to effort to develop gene-based methods to select for Calm, stress-tolerant, immune-competent, disease-resistant cattle. The biostimulatory effect of rams in ewes accelerated resumption of luteal function during the transition into the breeding season. More importantly, this effect appears to include changes in certain metabolic hormones that may be involved with the physiological mechanism of this biostimulatory effect during the transition into the breeding in sheep. Exposing primiparous, postpartum, anovular, suckled, beef cows to mature bull urine for 12 h daily does accelerate resumption of ovulatory activity but it does appear to improve artificial insemination pregnancy rates. Bull urine per se may not be the medium that carries the biostimulatory pheromone(s) that accelerates resumption of ovulatory activity in the bovine. However, bull urine may carry an additional pheromone(s) that is associated with fertility in the bovine. Peinatal exposure of male rats to progesterone or RU486 decreased initial sexual behaviors and numbers of serotoli cells. Differences in gene expression in LP rams as identified by micro-array analysis were not confirmed by quantitative PCR. Further analysis may reveal a panel of genes that may help identify LP rams but no overt changes were identified. Objective2. Develop and improve reproductive technologies to enhance sustainability of production systems for domestic ruminants. Estrus, hormones, estrous synchronizations and AI: The 5 d CO-Synch + CIDR program for timed results in an 11 percentage point increase in timed AI pregnancy rate relative to the traditional 7 d program. The requirement for 2 injections of can be circumvented by giving two doses of PGF at the time of CIDR withdrawal. This program provides a substantial increase in timed AI pregnancy rate and similar convenience to the traditional 7 d program. Pregnancy rates in ewes were improved by administering 10 mg of PGF2a at CIDR removal. Exposing heifers to bulls during an estrus synchronization protocol that included a CIDR for 14 d, followed 18 d later with PGF2± (PG), and, timed AI (TAI) and GnRH can substantially improve AI pregnancy rates. It appears that combining bull exposure with a estrus synchronization protocol that include controlled internal drug release devices (CIDR) for 14 d, followed 18 d later by PGF2± (PG), and, timed AI (TAI) and GnRH would be beneficial for improving reproductive performance of yearling beef heifers. Thus, inclusion of a bull in progesterone-based synchronization protocol would be an important, sustainable management practice to improve reproductive efficiency in beef cattle herds. The ovine adrenal gland has been reported to be a source of extragonadal inhibin. Present findings provide evidence that in wether lambs extragonadal inhibin does not exert a significant inhibition of FSH secretion. Cows culled from the herd at different ages may have altered follicular gene expression which is not optimal for follicular development and may affect fertility of that animal. Recombinant single chain analogs of ovine gonadotropins can be used to induce a superovulatory response in sheep and may have broader application in ovulation induction protocols. Puberty: Sex of lamb, type of birth, and age should be considered when evaluating serum hormone profiles in rapidly growing lambs. Scrotal circumference is highly correlated with dam milk production, but direct preweaning gain is only lowly correlated with scrotal circumference and postweaning gain is not correlated with scrotal circumference. Thus, how bulls are fed during development postweaning should not affect sperm production. No in utero effects on age of puberty or any spermatozoa morphological characteristics were detected indicating that purebred breeders should not realize a benefit in fertility of bulls by over conditioning their cows. Immunocastration The GnRH vaccine has proven to be an effective means of contraception in elk but it does not interfere with pregnancy existing at the time of immunization. GnRH antibodies are effectively transferred to the neonate via colostrums, but they do not interfere with normal development/function of the hypothalamic-pituitary gonadal axis in the offspring. Disease, immune function: Maternal thyroid deficiency during pregnancy might reflect a pregnancy with a PI fetus for BVD and could compromise fetal development and maternal health An experimental inoculum using infected splenocytes from SCID mice was used successfully to induce abortion in susceptible cattle. This method of transmitting infection to cattle is a quantifiable and predictable method of infection that has produced EBA induced abortions consistently through the past 2 years. Data generated from the past 2 years has lead to the production of a murine origin inoculum that that is being developed for use as a "stop gap" method of vaccination and protection in susceptible heifers and pregnant cows. The first year using this vaccine has shown it to be safe (none of the vaccinated heifers became ill or had abortions induced) and effective. Vaccinated animals did not abort when challenged by experimental infection as did the control group of animals. The development of a safe and effective method of protecting cattle from EBA is necessary for producers raising cattle in EBA endemic area of the West. It will also be of utility (or even more so) for producers outside of the area were EBA is known to occur that plan to move their animals into endemic regions in order to utilize new pasturing opportunities. It appears that even up to 3 year following an abortion cattle maintain enough immunity against the agent of EBA to protect against subsequent infection. The upcoming final year of this study will be used to expand the numbers of animals test to see if immunity 3 years post infection is consistently strong enough to protect cows from subsequent abortion. This information will be of use to cattle producers because it can potentially give them guidelines for determining which cattle to breed for the following year and help producers utilize pasture resources in pajahuello tick endemic areas with greater confidence. Behavior and Stress: Plasma ghrelin concentrations are differentially regulated in heat-stressed vs. pair-fed dairy cattle during hyperinsulinemia-euglycemia. The identification and use of high-sexually performing rams with desired genetic traits is a requisite to the timely incorporation of those traits into a flock. High-sexually active rams appear to exhibit more exploratory-type behaviors than low-sexually performing rams. The adrenal cortex stimulation test does not identify feed efficient cattle. Exit Velocity (EV; the rate at which calves exit from a handling chute) is a useful, viable and objective indicator of temperament classification of beef calves. The increased cost associated with the potential for more excitable or temperamental cattle to injure themselves or workers and to damage facilities, in addition to the negative effects of temperament on growth, immunity and carcass characteristics, motivates producers to select against more temperamental cattle. Although temperamental calves may be objectively identified as early as 21 days of age, temperament may be more easily identified prior to weaning by use of the Exit Velocity trait. This new management technique may enhance the ability of producers to select against temperamental animals. Transportation per se may not be as stressful as previously presumed, and it may be that the handling, loading and unloading of cattle may be more stressful than the transportation event itself. This knowledge may impact procedures to transport cattle as well as impact subsequent product quality. Selection pressure against temperamental cattle results in beef with improved sensory traits which should lead to increased consumer satisfaction which would permit Texas to have a greater market share of the meat industry. Pregnancy: Use of several parameters (especially ultrasound and estrous detection) after breeding allows for early diagnosis of pregnancy (d 19) which should improve our understanding of (and prevention of) early embryonic losses and may yield identification of blood components capable of accurately identifying pregnancy establishment. Nutrition: Selecting beef heifers for low residual feed intake (RFI), a feed efficiency trait, results in earlier return to estrus and improved conception rate when forage conditions are adequate. Plasma insulin-like growth factor-I (IGF-I) is not a physiological marker for feed efficiency in Brahman cattle.

Refereed Journal Publications: Alexander, B.M., P. Singh, K.J. Austin, R.R. Cockrum, K.M. Cammack, B.W. Hess, G.E. Moss, P.W. Nathaneilsz, and S.P. Ford. 2009. Effect of maternal fatness on fetal steroids and semi-quantitative real-time PCR expression of receptor genes in sheep. Anim. Reprod. Sci. 116:58-64. Epub 2008 Dec 24. Arndt, W. J., A. J. Holle, M. L. Bauer, J. D. Kirsch, D. E. Schimek, K. G. Odde, K. A. Vonnahme. 2009. Effects of progesterone supplementation on embryonic survival in dairy cows. Canadian Journal of Veterinary Research. 73:271-274. Araujo, R. C., Pires, A. V., Susin, I., Mendes, C. Q., Rodrigues, G. H., Urano, F. S., Ribeiro, M. F., Oliveira, C. A., Viau, P., and Day, M. L. 2009. Postpartum ovarian activity of Santa Ines lactating ewes fed diets containing soybean hulls as a replacement for coastcross (Cynodon sp.) hay. Small Ruminant Research 81:126-131. Arreguin-Arevalo J.A., R.L. Ashley , E.R. Wagenmaker , A.E. Oakley, F.J. Karsch, and T.M. Nett. 2010. Membrane-initiated actions of estradiol (E2) in the regulation of LH secretion in ovariectomized (OVX) ewes. Reprod Biol Endocrinol. 8:40doi:10.1186/1477-7827-8-40. Arellano-Rodriguez, G., C. A. Meza-Herrera, R. Rodriguez-Martinez, R. Dionisio-Tapia, D. M. Hallford, M. Mellado, and A. Gonzalez-Bulnes. 2009. Short-term intake of B-carotene-supplemented diets enhances ovarian function and progesterone synthesis in goats. Anim. Physiol. and Anim. Nutr. 93: 710-715. Arroyo, J.A., R.V. Anthony, T.A. Parker and H.L. Galan. 2010. eNOS, NO, and the activation of ERK and AKT signaling at mid-gestation and near-term in an ovine model of intrauterine growth restriction. Syst. Biol. Reprod. Med. 56:6273. Artac, R.A., R.M. McFee, R.A. Smith, M.M. Baltes-Breitwisch, D.T. Clopton, and A.S. Cupp. 2009. Neutralization of vascular endothelial growth factor antiangiogenic isoforms is more effective than treatment with proangiogenic isoforms in stimulating vascular development and follicle progression in the perinatal rat ovary. Biol. Reprod. 81:966-977. Arvisais E., X. Hou, T.A. Wyatt, K. Shirasuna, H. Bollwein, A. Miyamoto, T.R. Hansen, B.R. Rueda, and J.S. Davis. 2010. Prostaglandin F2alpha represses IGF-I-stimulated IRS1/phosphatidylinositol-3-kinase/AKT signaling in the corpus luteum: role of ERK and P70 ribosomal S6 kinase. Mol Endocrinol 24: 632-643. Aston K.I., G.P. Li, B.A. Hicks, B.R. Sessions, A.P. Davis, L.F. Rickords, J.R. Stevens, and K.L. White K.L. 2010. Abnormal Levels of Transcript Abundance of Developmentally Important Genes in Various Stages of Preimplantation Bovine Somatic Cell Nuclear Transfer Embryos. Cellular Reprogramming 12(1):23-32. Aston K.I., G.P. Li, B.R. Sessions, A.P. Davis, Q.A. Winger, L.F. Rickordsk, J.R. Stevens, and K.L. White. 2009. Global Gene Expression Analysis of Bovine Somatic Cell Nuclear Transfer Blastocysts and Cotyledons. Molecular Reproduction and Development 76:471-482. Baltes, M.M., R.A. Ten Broeck, R.A. Longfellow, R.M. McFee, D.T. Clopton, and A.S. Cupp. 2010. Neutralization of VEGF anti-angiogenic isoforms may regulate testis development. Reproduction. May 10 (Epub ahead of print). Bishop C.V., T. Filtz, Y. Zhang, O. Slayden, and F. Stormshak. 2008. Progesterone suppresses an oxytocin-stimulated signal pathway in COS-7 cells transfected with the oxytocin receptor. Steroids 73:1367-1374. Bott, R.C., R.L. Ashley, L.E. Henkes, A.Q. Antoniazzi, J.E. Bruemmer, G.D. Niswender, F.W. Bazer, T.E. Spencer, N.P. Smirnova, R.V. Anthony and T.R. Hansen. 2010. Uterine vein infusion of interferon tau (IFNT) extends luteal lifespan in ewes. Biol. Reprod. 82:725-735. Bridges, G. A., M. L. Mussard, C. R. Burke and M. L. Day. 2010. Influence of the length of proestrus on fertility and endocrine function in female cattle. Anim Reprod Sci. 117:208-215. Bryant, T.C., J.J. Wagner, J.D. Tatum, M.L. Galyean, R.V. Anthony and T.E. Engle. 2010.Effect of dietary supplemental vitamin A concentration on performance, carcass merit, serum metabolites, and lipogenic enzyme activity in yearling beef steers. J. Anim. Sci. 88:1463-1478. Burdick, N.C., J.P. Banta, D.A. Neuendorff, J.C. White, R.C. Vann, J.C. Laurenz, T.H. Welsh, Jr. and R.D. Randel. 2009. Interrelationships among growth, endocrine, immune and temperament parameters in neonatal Brahman calves. J. Anim. Sci. 87:3202-3210. Burdick, N.C. , J.A. Carroll, L.E. Hulbert, J.W. Dailey, S.T. Willard, R.C. Vann, T.H. Welsh, Jr. and R.D. Randel. 2010. Relationships between temperament and transportation with rectal temperature and secretion of cortisol ardepinephrine in bulls. Livestock Sci. 129:166-172. Burdick, N.C., B. Agado, J.C. White, K.J. Matheney, D>A. 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