Brief Summary of Minutes of Annual Meeting

Chair Andy Roberts called the meeting to order at 8 a.m. on May 22, 2006. Participants were introduced, minutes from the last meeting approved, and appreciation expressed to Dennis Hallford for arranging meeting facilities and food. Colin indicated that the W112 renewal was approved by the Western region without any requests for additional information or revisions. Colin also indicated that this was his first-ever experience with a renewal project that made it through the review process without any requests for modifications. He also expressed thanks to the writing team for crafting a project that was in good shape. The new number for the project will be W1112 (number was changed after the meeting). The official start date of the W1112 project is October 1, 2006 and it will terminate on September 30, 2011. Deb Hamernik discussed the new W1112 five year plans and reminded participants to pay attention to assurance statements when submitting their CRIS projects at their institutions. Valid approval for the use of recombinant DNA or animals (with approval dates) must be included on the Assurance Statement. There must be one year remaining on the IBC and IACUC approvals so that studies can be initiated during the first year of the project. On the CRIS AD-416 form, enter the objectives for the W1112 project rather than your own individual station objectives. Participants are encouraged to provide more information regarding impact statements. Impacts are defined as: measurable changes in economic social, health or environmental conditions to the discipline. Potential impacts can be discussed for basic projects. She also indicated changes in personnel within USDA CSREES. Applications to NRI must adhere to the published guidelines (budget caps, pages limits, etc.) or they will be returned without review. SF424 R&R application forms are now going to be used across the federal government. These forms are available on the CSREES website and will be used for NIH, NSF, DOE and NRI. Electronic submission of applications to NRI and SBIR will be required Oct 1, 2006 through grants.gov. Deb presented Jim Berardinelli an award because he was the first individual to submit an NRI application electronically through grants.gov. He was funded so it was in fact a success story. Congrats Jim!!! Business items included the unanimous election of Dean Hawkins as Member at Large and selection of Fort Collins, Colorado for the next W1112/W112 meeting on May 21-22, 2007. Tod Hansen was selected to lead the organization and arrangement of accommodations for the 2007 meeting. Announcements and news from individual stations were given. There was no discussion of old business. Individual state announcements occurred with members talking about events, job openings, etc that had happened since our last meeting. The remainder of the meeting was dedicated to presentations of individual reports, and identification of collaborative projects for the ensuing year. A lengthy discussion occurred determining how we can translate the research our group is currently involved in to the producer or producer-oriented groups. We discussed having a day symposia for producers, veterinarians, extension educators, etc on either nutritional effects on reproduction, oocyte quality, heifer development, etc. The meeting adjourned at noon on May 23, 2006. Over several subsequent e-mails the W1112/W112 group decided to have the 2007 meeting in Fort Collins, CO from June 5-6, 2007. The meeting date was changed to be prior to the Beef Improvement Federation (BIF) meeting that will be held from June 6-9th. We also would add a 4 hour session directed to the BIF membership and producers at the end of the W1112/W112 meeting that would address fetal development. We would close our meeting at noon on June 6th and then start the special session at 1pm. Six potential speakers were identified from this group. Tod Hansen and Terry Nett were selected from the membership to organize this special session.

Objective 1. Investigate molecular, cellular and endocrine mechanisms that limit or control reproductive efficiency in domestic animals. Estrous cycle Physiology/ CL function 1.) Intraluteal administration of meloxicam, a COX-2 inhibitor, increased the length of the ovine estrous cycle and length of time at the end of the luteal phase for serum progesterone to decrease to less than 1 ng/mL. 2.) Steady-state concentrations of mRNA encoding ODC and eNOS vary during the ovine luteal phase and are altered in response to PGF2a. 3.) Antisera that target defined epitopes on bioactive peptides may prolong the functional life of the peptide and thereby, enhance the physiological response. Therefore, these bioactive peptides may be useful in superovulation, synchronization protocols, or experiments to determine ovarian function. Hypothalamic-Anterior Pituitary Gland Physiology 1) Microarray analysis identified several genes that exhibited differential levels of expression in anterior pituitaries that were associated with the transition from anestrous to cycling status in postpartum cows. The roles that many of these genes have in regulating reproductive function have not been previously considered, and several of the transcripts represent as of yet uncharacterized genes. Thus, further verification of these results will undoubtedly result in the identification of novel mechanisms involved in regulating reproduction. 2) Identification of sheep fetal brain tissues expressing steroid receptors provides valuable information as to the roles of gonadal steroids in regulating sexual differentiation of the brain and potential causes for unique adult sexual behaviors of rams. 3) Recombinant gonadotropins can be used to induce precocious puberty in sheep and may have broader application in ovulation induction protocols. 4) The fact that estradiol receptors associated with the plasma membrane modulated numbers of GnRH receptors may provide the opportunity to develop novel treatments to regulate pituitary sensitivity to GnRH without influencing other cellular mechanisms regulated by the nuclear estradiol receptor. 5) Identification of a functional G-protein coupled receptor for progesterone in sheep provides a putative explanation for the rapid effects of progesterone on cellular activity. A better understanding of the physiological role of this receptor may enhance our ability to regulate individual biological actions of progesterone. 6) The KiSS-1 peptide was shown to modulate LH secretion in the ewe exclusively by modulating secretion of GnRH without any effect on the pituitary gland. Effects of KiSS-1 were not influenced by presence of estradiol and progesterone. This peptide may provide a novel means of regulating LH secretion independent of the actions of estradiol and progesterone. 7) Antisera that target defined epitopes on bioactive gonadotropin peptides may prolong the functional life of the peptide and, thereby, enhance the physiological response. Ovarian Physiology. 1) Ovulation of smaller ovulatory follicles results in lower pregnancy rates in beef cows and heifers. All mechanisms by which ovulation of smaller follicles reduces pregnancy rates have not been elucidated, but at least a portion of this reduction in fertility appears to be due to reduced subsequent progesterone synthesis. It makes sense that endogenous estrogen secretion is also indicative of follicular maturity and subsequent progesterone synthesis. Results of these studies should improve the ability to develop ovulation synchronization protocols and management practices that ensure fertile ovulations. 2) Treatment of bovine cortical cultures with VEGFR-TKI (8uM) inhibited follicle activation and resulted in more follicles at earlier stages of development. Thus, inhibition of VEGF signal transduction can inhibit follicle progression. This is similar to data in the rat. Using rat perinatal ovaries both VEGF164 (recombinant) and an antibody to the inhibitor VEGF isoforms had similar effects with increases in vascular density and follicle progression to advanced stages of development. While inhibition of the VEGFR2 receptor surprisingly increased vascular development but inhibited follicle progression. Understanding the regulation of VEGF may allow for better manipulation of follicle progression and development in domestic livestock. 3) An inhibitory isoform, VEGF164b has been isolated and sequenced in the bovine. Expression of mRNA for angiogenic isoforms VEGF164, 120 and inhibitory isoform VEGF164b are differentially regulated prior to the LH surge. Both angiogenic isoforms are reduced while VEGF164b is increased at 18 hrs after prostaglandin. Only VEGF164 is increased after the LH surge. Understanding why these isoforms are expressed prior to and after the LH surge will allow us to manipulate ovulation and potentially also alter formation of the CL. 4) Granulosal cell CART mRNA and follicular fluid concentrations of the CART peptide are negatively associated with health status of bovine follicles collected at multiple defined stages of follicular development. 5) CART treatment of bovine granulosal cells inhibits FSH induced influx of extracellular calcium in a Go/Gi dependent manner suggestive of a role for regulation of [Ca2+]i in CART signal transduction. 6) Alternative Expression of FSH Receptor Transcripts: We have previously determined that alternate transcripts of the FSH receptor are produced differently at important stages of follicular development in the cow. To better understand the precise role of each form of the FSH receptor on granulosa cell differentiation and development, stable granulosa cell lines have been obtained with produce either FSHR-1 or FSHR-3, and no other forms of the FSHR. Using these cell lines, a series of experiments is planned to characterize patterns of expression of both mRNA and protein for each of these forms of the FSHR, as well as to validate techniques to enhance our ability to detect each form of the FSHR from bovine tissues or primary cultures of granulosa cells. Oocyte-embryo Physiology. 1) JY-1 (a novel oocyte/embryo specific gene) mRNA transcripts present in early embryos are of maternal origin and appear to be required for early embryonic development. A cDNA encoding for a putative human JY-1 ortholog has been cloned from human ovary and ES cell RNAs. 2) 1756 RNA transcripts of oocyte and cumulus cell origin representing genes co-regulated and differentially regulated within the two cell compartments during meiotic maturation were identified. 3) A novel algorithm has been developed and applied to characterization of components of the oocyte transcriptome using data derived from 2 channel microarray experiments. 4) A negative association of cumulus cell cathepsin mRNA abundance with oocyte competence and development to the blastocyst stage has been established. Treatment with a cathepsin inhibitor during meiotic maturation enhanced blastocyst development. 5) Bovine/Porcine stem cell project: We have focused on inserting marker genes into pig umbilical cord cells with subsequent injection of cells into blastocyst-stage embryos. If these cells can integrate into various tissues of the developing embryo, it would confirm their identity as pluripotent stem cells. Testicular Physiology. 1) Immunoneutralization of inhibin holds potential as an approach to increase sperm production. Results of study one show that the magnitude of change in sperm production is directly proportional to the inhibin antibody titer achieved. Inhibin appears to be involved in the regulation of the number of primary spermatocytes. 2) Results of study two indicate that inhibin free "-subunit has little if any effect on pituitary LH secretion. Thus, the immunoneutralization of endogenous inhibin free "-subunit likely does not account for the decrease in LH secretion that occurs in ram lambs actively immunized against inhibin free "-subunit. This information will be used in the development of immunogens designed to stimulate gametogenesis. 3) Treatment of indifferent testes with VEGF164 increased vascular density by 57% (P < 0.05) and perturbed seminiferous cords similar to results with another isoform, VEGF120. We determined VEGF164b and 188b mRNA were not expressed in the testis during cord formation and testis morphogenesis, but both were present in the ovary at the same time point. Furthermore, mRNA for Flt1 is expressed in the ovary and not in the testis. We speculate that negative regulators of VEGF, VEGF b isoforms and Flt1, may inhibit mesonephric endothelial cell migration in the ovary where their absence allows for endothelial cell migration in the testis resulting in testis-specific vascular development. 4) VEGF treatment increased graft weight and the percentage of seminiferous tubule cross-sections with elongating spermatids at the time of graft removal. Cultured testis tissue grafts were smaller, and had fewer seminiferous tubules per graft. However, there was no difference in the percentage of seminiferous tubule cross-sections that contained any germ cell type between groups. These data indicate for the first time that bovine testis tissue can be manipulated to better support germ cell differentiation in grafted tissue. 5) We generated data demonstrating that both donor age and grafting period may be important factors regulating the maturation of bovine testis xenografts, indicating that intrinsic differences exist within testis tissue at these donor ages. These data provide the framework for further study of bovine spermatogenesis using ectopic testis xenografting. 6) Examination of gene expression in testis tissue grafts using real-time RT-PCR indicated that Sertoli cell specific genes were lower in 8 week donor tissue grafts compared to the other aged donors. Additionally, c-kit, a germ cell specific gene was nearly absent in grafted tissue. Microarray analysis of donor tissue identified several genes that are involved in angiogenesis or tissue growth that were differentially expressed in 2, 4, and 8 week old bovine testes. Expression of these genes (angiogenin, transgelin, thrombomodulin, early growth response 1, insulin like growth factor 2, and insulin like growth factor binding protein 3) decreased over time. This research identified several factors that could contribute to efficiency of ectopic testis tissue grafting. This knowledge could lead to exogenous treatements for improved ectopic testis grafting techniques. 7) Cryopreservation adversely effected growth, testosterone production, and germ cell differentiation of mouse grafts. Cryopreservation did not appear to influence growth, testosterone production, or germ cell differentiation in pig grafts. These data indicate that cryopreservation is a feasible method for long-term testis tissue storage prior to ectopic testis xenografting. 8) FSH and testosterone contribute to the spermatogonial stem cell niche to regulated stem cell biological activity. 9) Using DNA parent verification there are considerable differences in serving capacity of individual bulls in a multiple sire breeding herd and this variation is not correlated with scrotal size. Uterine-Placental-Fetal-Nutritional Interactions during Pregnancy. 1) More than one agonist may regulate caruncular endometrial PGE secretion for implantation. 2) In the ewe, the conversion of placentomes from A to D type increases their ability to delivery nutrients to the fetus, thereby protecting the fetus from intrauterine growth restriction. This conversion normally occurs during late gestation in response to the exponential growth of the fetus, but some animals have the ability to convert their placentomes during early gestation in the face of a maternal stressor such as undernutrition. This ability for early placentomal conversion appears to be related to pre-gestational ewe selection, and thus may be epigenetic in nature. These data suggest that the management system under which a ruminant female is selected can have profound effects on her ability to protect her fetus from intrauterine growth restriction in the face of maternal insults such as undernutrition. 3) Fetal intrauterine growth restriction resulting from undernutrition during the first half of gestation in ewes and cows have significant negative health effects on their offspring after birth, including insulin resistance, hyperphagia, obesity, decreased skeletal muscle mass, hypertension and infertility . These health, fertility and body compositional effects of early maternal undernutrition are not alleviated even if females are realimented during the second half of gestation, and are born at a normal birth weight. 4) It was demonstrated that concentration of estradiol during the preovulatory period did not alter total uterine IFNÄ content in pregnant heifers on d 15.5 of gestation but level of nuclear progesterone receptor mRNA in the uterine endometrium tended (P < 0.10) to be greater in females with greater estradiol concentrations during proestrus. 5) The presence of estrogen receptor in plasma membranes of the ovine endometrium can be used to examine the nongenomic rapid actions of estrogen during various stages of the estrous cycle and gestation as related to establishment and maintenance of pregnancy. Ability of progesterone to inhibit oxytocin-induced cellular function in transfected COS-7 cells may serve as a model system to further examine the mechanism by which this steroid acts to maintain pregnancy in ruminants. 6) Adaptive responses to fetal nutrient deprivation have a greater impact on fetal ²-cell development and function than other endocrine cell types within the pancreas. Insulin is an important anabolic hormone in the fetus and fetal ²-cells respond to nutrient concentrations during the second half of gestation; thus, making ²-cells important for coordinating the rate of fetal growth with the maternal nutrient supply to the fetus. We show that the disparity in fetal hypoglycemia and hypoxia increase over the final 30% of gestation, and the ²-cell responsiveness also decreases due to lower ²-cell mass and impaired islet physiology and metabolism. In addition the IUGR fetuses develop a greater sensitivity to insulin, which might be required to respond to the lower ²-cell responsiveness. These adaptations will promote fetal survive during nutrient deprivation, but if prenatal compensatory mechanism persist after birth they could result in complications in animal health, production, or market value. 7) Impaired placental function is the primary cause of fetal growth restriction. We have demonstrated that the development of functional placental insufficiency in sheep is associated with changes in placental expression of the insulin-like growth factors and their binding proteins, increased placental apoptosis, and differential changes in endothelial nitric oxide abundance in the cotyledon and umbilical artery. The observed changes help explain the failure of the placenta to develop normally, and why there is impaired oxygen and nutrient provision to the fetus. 8) Impaired fetal development has been associated with altered postnatal growth and health of the offspring. We demonstrated that prenatal androgen exposure not only reduces size at birth, but also results in changes in left ventricular size and concentration of endothelial nitric oxide and insulin receptor indicative of long-term physiological changes in adult sheep which may impact their health and longevity. Nutritional and metabolic effects in non-pregnant animals. 1) Short-term (5 d) feed deprivation prior to ram introduction during proestrus results in an approximate 30% reduction in lamb crop. Provisions need to be made to ensure that adequate feed resources are available during winter breeding in ewes to prevent feed deprivation that can occur as a result of adverse environmental conditions. 2) In reindeer, seasonal body weight changes and pregnancy status will be correlated with seasonal changes in plasma leptin, IGF-1 and GH and these data combined with retrospective data collected in 2002/03. 3) We have determined that nutrition may affect expression of VEGF isoforms in the ovulatory follicle. There was an increase in VEGF120 isoform in heifers fed whole soybeans for 167 days during heifer development that were aspirated 60 hours after synchronization with prostaglandin compared to controls. These differences were not correlated with changes in progesterone or estrogen. Also there was not difference in VEGF164 isoform. We are presently evaluating VEGF164b isoform. 4) Heifers which are more efficient in feed utilization, low relative feed intake, have less body fat than less efficient heifers. 5) Heifers with low relative feed intake reached puberty at similar rates as their less efficient (but fatter) herdmates. Behavior. 1) Both adrenal cortical and medullary function are influenced by temperament in response to typical animal handling procedures, including transportation. 2) Weaned calves with poor temperaments are less competent immunologically than those with calm temperatures. 3) Breed type of cattle influenced anatomic parameters of the hypothalamic-pituitary-adrenal axis whereas temperament influences endocrine function of the adrenal cortex and medulla. 4) Cattle with poor temperaments have reduced fat deposition compared with calm cattle. Objective 2. Develop and improve assisted reproductive technologies to enhance sustainability of production systems for domestic ruminants. Estrus, estrous synchronization and AI. 1) In reindeer, we found a difference in gestation length between the EARLY and LATE groups (P = 0.002) and a negative correlation (r = -0.754; P = 0.002) between gestation length and breeding date. There was no difference in the birth weight of the calves. 2) We are analyzing reindeer milk for %fat, %protein, %lactose and % solids-not-fat at 3 stages of lactation (15, 45, and 75 d post-calving) from 12 female reindeer. Milk component data from each cow will be correlated with their calfs growth rate and survival. 3) Use of Modified CIDR-b for Estrous Synchronization in Reindeer Bovine CIDR modifications (following manufacturers directions) produced systemic progesterone levels consistent with luteal phase progesterone and had 100% retention rate. Pregnancy rate following synchronization was 88%. 4) Radiotelemetric Estrous Detection in Reindeer. Placement of radiotelemetric transmitters lower on the females rump, coupled with shaving only the top 1.5 cm of hair, were tried in an effort to improve estrous detection in reindeer with this technology. 5) Modifications of the standard CO-Synch + CIDR estrous synchronization program that included: a. shortening the interval from the initial GnRH injection to CIDR withdrawal/PGF2± from 7 to 5 d, b. administering two PGF2± doses 12 h apart and c. increasing the interval from PGF2± to the second GnRH from 60 to 72 h; increased timed-AI pregnancy rates in postpartum beef cows by 13.3%. 6) Development of dominant follicles occurs early after calving and lack of follicular development does not delay return to estrus. It seems to be obligatory for most cows to have a silent ovulation with formation of functional luteal tissue before return to normal estrous cyclicity. 7) Early weaning at the start of a synchronized breeding season increased AI pregnancy rates and cow weights at the time of normal weaning. The magnitude of improvement in AI pregnancy rate is greater than has been reported with temporary calf removal (48 h) indicating the effects of permanent calf removal may be manifested through improved pregnancy maintenance. Early weaning may be a viable alternative to culling cows during periods of low forage production in semi-arid rangelands. We are evaluating the fertility of heifers developed under early and normal weaned strategies. 8) Liver and skeletal muscle progesterone concentrations did not differ from control values in ewes receiving CIDR for 5 or 14 d. Therefore, the levels of progesterone in CIDR treated ewes were within limits allowable for slaughter and increased the chances of CIDRs being approved for utilization in regulation of sheep estrous cycles. 9) Fenceline contact of cows with bulls can accelerate resumption of ovarian cycling activity but it is not as effective as close physical contact between cows. The nature of the biostimulatory effect of bulls appears to be related to the intensity of the pheromonal stimulation. Limiting the intensity of the biostimulatory stimuli of bulls directly limits that biostimulatory effect of bulls on primiparous cows. Thus, a major factor involved in the mechanism of the biostimulatory effect of bulls is intensity of the stimuli (frequency of exposure, duration of exposure, and quantity of stimuli) provided by bulls. 10) Continuous presence of bulls stimulates resumption of luteal activity and is coincident with increased cortisol concentrations. We hypothesize a possible association between adrenal activation and the biostimulatory effect of bulls in postpartum anestrus cows. This may be a critical mechanism for this effect. The biostimulatory effect of bulls alters the progesterone response in an ES protocol that included GnRH, PGF2±, GnRH and TAI, and that progesterone concentrations after the first GnRH injection may determine the success rate of GnRH-based ES protocols in primiparous beef cows. 11) Comparison of CIDR to MGA in a 7-11 Co-synch protocol with timed insemination. Previous research has shown that the 7-11 Cosynch protocol using melengestrol acetate (MGA) is effective in synchronizing beef heifers. This study compared a vaginal insert containing progesterone (CIDR) to MGA in the 7-11 Cosynch protocol on beef heifers. No difference in pregnancy rate was observed between the CIDR (46%) and MGA (47%) treatments. Puberty. 1) Attainment of puberty in early-weaned heifers can be hastened through feeding a high-concentrate diet from approximately 4 to 6.5 mo of age in a manner similar to that of heifers that are continuously fed the high-concentrate diet after early weaning. 2) Early postnatal concentrations of metabolic hormones are related to growth characteristics and may therefore be related to age at onset of puberty. 3) Determined that dried distillers grains has a positive effect on conception to AI when fed during heifer development and therefore is an effective method to provide protein to heifer development diets with no negative effects on reproductive performance. Pregnancy detection and maintenance. 1) Detection of low ISG15 mRNA levels in blood on days 17-25 provides an accurate indicator of cows that are not pregnant. This might be used as a management tool to help identify nonpregnant cows 18 days following AI so that they can immediately be re-synchronized and inseminated. 2) The results of these experiments suggest that if intraluteal concentrations of progesterone can be maintained during the period of normal luteolysis that it should be possible to prevent luteolysis and perhaps reduce the 20-35% early embryonic wastage that is attributable to faulty mechanisms associated with maternal recognition of pregnancy. 3) Pregnancy induces many of the same blood and endometrial genes and several of these are involved with interferon and antiviral responses. Many pathways are also uniquely induced in blood, when compared to the endometrium, and may function in maintaining early pregnancy in the cow. 4) The decidualizing uterine stromal compartment, in addition to the ovarian corpus luteum and placenta, is a site of steroid synthesis during early gestation, and the embryo provides signaling cues to tightly regulate steroid biosynthesis at the maternal:embryo interface. Similar mechanisms probably occur in the cow. 8. Maternal nutrient restriction alters gene expression in the fetal heart, which may affect sensitivity of the steer heart to stress by 15 months of age. Differential programming of right ventricular gene expression in the fetus during early gestation may be detrimental to animal health, particularly at high altitude. Immunocastration. One of the problems with development and approval of a sterilization vaccine for cattle has been the stigma against Freunds complete adjuvant. We have evaluated a bacterial DNA immunostimulant with an ovalbumin-LHRH fusion protein and found the bacterial DNA to be superior to Freunds in stimulating LHRH antibody production. Disease. 1) Persistent infection with BVDV results in antiviral responses in blood cells which includes induction of type 1 interferon-induced genes, chemokine-mediated immune responses and bone remodeling with a concomitant suppression of extracellular remodeling, adhesion and T-cell-mediated responses. 2) Development of a single or multiplexing diagnostic for calves that are persistently infected with BVDV is probable. Effort. Participants in W-112 have published 68 refereed journal articles, 2 book chapters and one genbank sequence during the past reporting period, many of which were derived from collaborative efforts initiated through the W1112/W112 Regional Research Project. Collaborative projects planned for the coming year are listed below. Collaborative Efforts for the Coming Year: Collaborators: Alaska, Montana. Project. Determine Glucocorticoid levels in reindeer. Collaborators: Alaska, New Mexico. Project. Determine Thyroid hormone concentrations in blood from reindeer. Collaborators: Alaska, Missouri. Project. Determine Leptin- IGF-1 and GH in blood from reindeer experiments. Collaborators: Alaska, Montana State. Project. Evaluation of cortisol concentrations in reindeer during photoperiod cycle. Collaborators: Arizona, Michigan. Project. Determine the impact of placental insufficiency and IUGR on ovary development to elucidate the potential impact in female reproductive performance. Collaborators: Colorado, Minnesota. Project. Examine the ability of alpha subunit of inhibin to regulate LH secretion in sheep. Collaborators: Colorado, New Mexico. Project: Evaluate the role of luteal PGs in demise of the CL in cattle and sheep. Collaborators: Colorado, Wyoming Project: Undernutrition in fetal development. Collaborators: Colorado, Wyoming, Ohio, Montana. Project: Evaluate blood and endometrial ISG15. Collaborators: Colorado, Penn State Project: Pregnancy induced gene expression in bovine and ovine blood and endometrium. Collaborators: Colorado, Idaho, Missouri, Ohio, USDA (Miles City). Project: Examine ISG15 mRNA in blood from pregnant cows. Collaborators: Colorado, Texas. Project: Examine ISG15 mRNA in blood from pregnant deer and Brahman cows. Collaborators: Hawaii, Texas. Project: Growth factor influences on bovine caruncular secretion of PGE1. Collaborators: Hawaii, Colorado. Project: Regulation of mRNA for enzymes in the PG synthesis pathways in sheep uterus. Collaborators: Idaho, Ohio, Colorado Project: Determine preovulatory estradiol and uterine expression of ISG-15, MX and other proteins. Collaborators: Kansas, Nebraska Project: Evaluate FSHR and VEGF expression in ovary and testis cultures. Collaborators: Michigan, Wyoming. Project: Determine factors affecting oocyte quality. Collaborators: Montana State and USDA (Miles City, Montana). Project: Utilization of the biostimulation effect of bulls to improve estrus synchronization response and fixed-time AI in postpartum cows. Collaborators: Nebraska, Arizona. Project: Determine effects of nutrient restriction in lambs on testis development and VEGF production. Collaborators: Nevada, Wyoming. Project. Differential gene expression in placentomes as they progress from type A to type D. Collaborators: Nebraska, WSU. Project. Role of VEGF in bovine testis grafts. Project. Spermatogonial germ cell transplantation Collaborators: Nebraska, Oregon. Project. Evaluate vascular development in fetal gonads from sheep. Collaborators: Ohio, Michigan State. Project. Examine CART expression in granulosa cells of follicles following estradiol induced atresia. Collaborators: Oregon, Wyoming. Project. Collaborate on MOR and FOR ram analysis. Collaborators: Texas, USDA (Miles City). Project. Feed efficiency in developing heifers Collaborators: Texas, USDA (Miles City, Montana). Project. Examine amygdallas characteristics associated with high and low EV bulls and heifers after puberty. Collaborators: WSU, USDA (Miles City). Project. Determine differences in pituitary gene expression in cycling and postpartum anestrus cows. Project. LHRH immunization. Collaborators: USDA (Miles City), Ohio. Projects. Effect of early weaning on age at puberty, 5 vs 7 d Co-synch + CIDR, estrogen and fertility at timed AI. Collaborators: USDA (Miles City), California. Project. Evaluation of gonadotropin analogs. Collaborators: USDA (Miles City), Colorado. Projects. Early Pregancy gene expression related to ovulatory follicle size/maturation. Progestin/Progesterone affinity for different progesterone receptors. Collaborators: USDA (Miles City), Missouri. Projects. Reciprocal ET of cows ovulating large or small follicles into recipients ovulating L/S follicles. Collaborators: Washington, Nevada. Projects. Gene expression in developing testis. Collaborators: Washington, Minnesota Projects. Inhibin A- immunized rams and effects on spermatogenesis. Collaborators: Wyoming, California, Nebraska, Arizona. Teaching Collaborations. Sharing of notes, lectures, case studies for Mammalian Endocrinology course.

Refereed Publications: 1) Adams TE. Using gonadotropin-releasing hormone (GnRH) and GnRH analogs to modulate testis function and enhance the productivity of domestic animals. Ani. Repro. Sci. 88:127-139. 2005. 2) Alexander, B.M., Z. Kiyma, M. McFarland, E.A. Van Kirk, D.M. Hallford, D.E. Hawkins, K.K. Kane, and G.E. Moss. 2006. Influence of short-term fasting during the luteal phase of the estrous cycle on ovarian follicular development during the ensuing proestrus of ewes. Anim. Reprod. Sci. In Press. 3) Alexander, B.M., R.H. Stobart, W.C. Russell, K.I. ORourke, G.S. Lewis, J.R. Logan, J.V. Duncan and G.E. Moss. 2005. The incidence of genotypes at codon 171 of the prion protein gene (PRNP) in five breeds of sheep and production traits of ewes associated with those genotypes. J. Animal Sci. 83:455-459. 4) Anil L, Anil S, Deen J, Baidoo S, Morrison R, Wheaton J, 2005. Evaluation of well-being, productivity, and longevity of pregnant sows housed in groups in pens with an electronic sow feeder or separately in gestation stalls. Am J Vet Res 66:1630-1638 5) Arreguin-Arevalo, A. and Nett, T.M. 2005. Nongenomic action of 17b-estradiol as the mechanism underlying the acute suppression of secretion of luteinizing hormone. Biol. Reprod. 73:115-122. 6) Arreguin-Arevalo, J.A. and Nett, T.M. 2006. A nongenomic action of 17b-estradiol as the mechanism underlying the acute suppression of secretion of luteinizing hormone in ovariectomized ewes.. Biol. Reprod. 74:202-208. 7) Barry, J.S., M.L. Davidsen, S.W. Limesand, H.L. Galan, J.E. Friedman, T.R.H. Regnault, W.W. Hay Jr. Developmental Changes in Ovine Myocardial Glucose Transporters and Insulin Signaling following Hyperthermia-Induced Intrauterine Fetal Growth Restriction. Experimental Biology and Medicine 231(5):566-75. 2006. 8) Bettegowda, A., O.V. Patel, J.J. Ireland and G.W. Smith. 2006. Quantitative analysis of messenger RNA abundance for ribosomal protein L-15, cyclophilin-A, phosphoglycerokinase, b-glucuronidase, glyceraldehyde 3-phosphate dehydrogenase, b -actin and histone H2A during bovine oocyte maturation and early embryogenesis. Molecular Reproduction and Development 73:267-278. 9)Bishop, C.V. and F. Stormshak. 2006. Nongenomic action of progesterone inhibits oxytocin-induced phosphoinositide hydrolysis and prostaglandin F2a secretion in the ovine endometrium. Endocrinology 147:937-942. 10) Bott RC, McFee, R M, DT Clopton, CF Toombs and AS Cupp. Vascular Endothelial Growth Factor (VEGFA) is involved in vascular development and seminiferous cord formation during testis morphogenesis in the rat. 2006; 75: 56-67; Biol Repro. 11) Bott RC, DT Clopton, RM McFee and AS Cupp. Kinase Domain Receptor (Kdr) expression is present in sex specific vascular patterns in the developing gonads: Role for VEGF in regulation of testis specific vasculature and spermatogenesis. (Submitted to Biol Repro. In review). 12) Burke, C. R., H. Cárdenas, M.L. Mussard, C.L. Gasser, and M.L. Day. 2006. Steroidogenic changes and expression of messenger RNA encoding steroidogenic enzymes, gonadotrophin receptors and cell-death signalling in the dominant ovarian follicle during oestradiol-induced atresia in cattle. Anim. Reprod. Sci. (In review). 13) Carlin, R.W., R.L. Sedlacek, R. R. Quesnell, F. Peirucci-Alves, D.M. Grieger and B.D. Schultz. PVD9902, a porcine vas deferens epithelial cell line that exhibits neurotransmitter-stimulated anion secretion and expresses numerous HCO3 transporters. Accepted, American Journal of Physiology-Cell Physiology, Dec 20, 2005. 14) Corcoran, D., T. Fair, D. Rizos, O.V. Patel, G.W. Smith, S. Park, P.M. Coussens, J.J. Ireland, M.P. Boland, A.C.O. Evans and P. Lonergan. 2005. Differential developmental Competence of in vivo compared to in vitro cultured bovine embryos is associated with differential gene expression and a suppression of transcription. Reproduction 131:651-660. 15) Dong, F., S.P. Ford, C. Fang, M.J. Nijland, P.W. Nathanielsz and J. Ren. 2005. Maternal undernutrition during early to mid-gestation up-regulates cardiac insulin-like growth factor (IGF) receptors associated with ventricular hypertrophy in fetal sheep. Growth Hormones & IGF Research 15:291-299. 16) Du, M., M.J. Zhu, W.J. Means, B.W. Hess and S.P. Ford. 2005. Nutrient restriction differentially modulates mTOR signaling and ubiquitin-proteasome system in skeletal muscle of cows and their fetuses. J. Anim. Sci. 83:117-123. 17) Echternkamp, S.E., K.A. Vonnahme, J.A. Green and S.P.Ford. 2006. A comparison of concentrations of vascular endothelial growth factor and pregnancy-associated glycoproteins in maternal blood between cows gestating single versus twin fetuses. 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