Brief Summary of Minutes of Annual Meeting

Minutes of the W-2171 Multi-State Project Meeting February 19-20, 2010 Animal Reproductive Biology Laboratory Colorado State University Fort Collins, CO Attendees (Station): Milan Shipka (AK) Rick Rorie (AR) Charles Rosenkrans (AR) Gerrit Bouma (CO) George Seidel (CO) Quinton Winger (CO) Rebecca Krisher (IL) Ken Bondioli (LA) Erdogan Memili (MS) Brett White (NB) Charlotte Farin (NC) Mark Mirando (USDA) Tom Bunch (UT) Chris Davies (UT) Clay Isom (UT) Qinggang Meng (UT) Meeting was called to order by project Chair, Char Farin, at 8:30 a.m. Attendees made brief introductions, followed by introduction of our seminar speakers by George Seidel. Gerrit Bouma and Quinton Winger presented Pluripotency in Trophoblast Stem Cells and Germ Stem Cells. Following the informative presentations, attendees asked questions and discussed the biology and potential future for stem cells. Mark Mirando presented AFRI: Update about the USDA Reorganization and Comments on Preparing Competitive Proposals for AFRI Submission. Mark distributed a National Institute of Food and Agriculture Factsheet that described the mission, leadership, and structure of the newly formed organization. In addition, he summarized the 2009 funding for AFRI, and explained the anticipated changes for 2010 NIFA funding for research, education, and extension. Since the RFA had not been released, Mark did not have the liberty to address specifics for programs; however, he was very helpful in explaining the obvious and subtle changes to funding guidelines and answered as many questions as possible. After a lunch break, the committee started station reports. The format for station reports this year was a 15 minute presentation followed by 5 minutes for discussion of potential collaboration. Most of the stations prepared an electronic presentation summarizing their work. Station reports concluded at 5:15 p.m. and meeting was suspended for the evening. Committee meeting was reconvened Saturday at 8:17 a.m. Administrative representative, Milan Shipka, addressed our group and indicated the importance of developing genuine collaboration and continued productivity. The committee then discussed each of the two project objectives. Objective 1: Oocyte quality was discussed and the following questions/observations were made: Why are in vitro matured oocytes not the same as in vivo matured oocytes? We were reminded that blastocysts are not the endpoint, rather live births are the endpoint! gene expression could/should be evaluated, possibly via miRNA and gene chips oocyte quality questions: small vs. large follicles; prepubertal vs. postpubertal vs. lactating females secreted items: miRNA, MHC proteins, cytokines... How does aneuploidy impact our results? Post-natal death appears to be increased for bulls resulting from x-sorted sperm. How do mosaic cells impact development? Tool development was discussed. Possibilities included: specific chromosome paint(s), deep sequencing PCR with 2-3 markers per chromosome, expressed tag sequencing... ...its not just an egg problem, we need to continue evaluating sperm... One approach for Objective 1 could be the evaluation of aneuploidy on in vivo and in vitro losses of oocytes and sperm. Assays would include: metabolic load, blastocyst percentage and quality, survival after cryopreservation, and offspring. Does cryopreservation induce aneuploidy? What about culture conditions? George Seidel agreed to lead the group in developing a funding proposal related to this topic. Char Farin and Rebecca Krisher agreed to manage the writing process. Objective 2: Reprogramming was the primary topic for discussion and the following questions/observations were made: How successful are induced pluripotent stem cells? Relative to cloning? How do we improve genetic reprogramming? How does the oocyte conduct genetic reprogramming? Oocyte does considerable erasing of alterations (i.e. demethylation&), but not all modifications need to be erased. Which need to be modified? transcriptional profiling could lead to answers for consideration ...lack of cloning success is probably not a genetic defect, rather an epigenetic defect... Business meeting was officially started at 10:20 a.m. Committee unanimously decided to hold the 2011 meeting in conjunction with IETS meeting in Orlando FL. Committee Chair for 2011, Charles Rosenkrans, agreed to check with IETS to determine the date and site preparation. First priority would be Friday January 7th from 8:00 to 5:00 without a guest seminar. Erdogan Memili was unanimously selected as the incoming project secretary. Committee members thanked George Seidel for hosting our committee and for providing snacks and drinks for breaks, and for arranging tours of horse and laboratory facilities. In addition, committee thanked University of Arkansas Animal Science department for providing the bound annual committee report. Utah committee members offered to host the committee for the 2013 meeting, and that invitation was unanimously accepted. Project Chair, Char Farin, agreed to write thank you letters to our guest speakers (Drs. Bouma and Winger). Meeting was adjourned at 10:45 a.m. Respectfully submitted, Charles Rosenkrans, Jr. 2009-2010 Secretary

Objective 1: Understand the biology and underlying mechanisms of gamete development, fertilization, and embryogenesis Results indicate the length of the MGA-SelectSynch estrous synchronization protocol can be reduced and CIDR progesterone inserts can be used instead of MGA without adversely affecting estrous response in treated cows. Organic mineral supplementation improves some semen quality characteristics in bulls during hot weather but additional studies are needed to determine whether this improvement translates into improved pregnancy rates. Bovine heat shock protein 70 promoter and coding sequence are polymorphic. Those polymorphisms are associated with calving rates of beef cows. Decreased fertilizability was detected in oocytes retrieved from gilts exposed to elevated ambient temperature for four days, corresponding to a typical weaning to estrus interval. We were unable to improve in vitro embryo production using cAMP regulators under the conditions tested; more research is required. We did find a number of differences in microRNA content at different stages of oocyte maturation/zygote production. These provide a starting point for manipulating these molecules during in vitro embryo production. We found that a GnRH-induced LH surge does not result in GVBD of oocytes in vivo under luteal concentrations of progesterone. A low Na+ low Ca++ base medium did not improve bovine vitrification success. We developed a simplified method of cooling bovine embryos for vitrification. Studied expression profiles of in vitro produced and in vivo produced bovine fetuses at Days 90 and 180 of gestation in two important organs: liver and placenta. The accomplishment of this study was the development of homogeneous embryonic stem cell derived clonal germ cell lines that could be continually propagated and differentiated into haploid cells. This in vitro germ cell culture system will allow for in depth study of the mechanisms orchestrating germ cell development and problems in livestock germ cell differentiation that hinder production. Leptin and glucose may interact to regulate oocyte nuclear maturation in obese and/or diabetic individuals. The glutaredoxin pathway is involved in oocyte developmental competence in pigs. Microfluidic devices provide the opportunity to culture oocytes and embryos individually, with development equal to that of standard culture drops. Abnormal fetal and placental development and differential expression of imprinted genes in the ovary and brain of aged mice suggests that epigenetic regulation of oocyte and fetal development is impaired with advanced maternal age. It has been established that Angus semen processed and frozen as early as 1960 and stored in LN is still viable and produces pregnancies at rates similar to semen frozen as recently as 2003. It was demonstrated that animal temperament measured by chute scores and chute exit velocity does not affect subsequent AI pregnancy rates in beef cattle. It was demonstrated that the addition of GnRH to a 14 day estrous synchronization protocol in White-tailed deer did not result in higher pregnancy rates. Transrectal ultrasonography in White-tailed dear as early as 73 days post-insemination allows for the differentiation of pregnancies derived from AI and intact bucks. It was demonstrated that laser assisted hatching in frozen-thawed bovine embryos does not increase the number of embryos hatching in vitro or establishing pregnancy upon transfer to recipients. Pluripotent-responsive, bovine NANOG promoter-GFP and tetracyline-responsive bovine OCT4 and SOX2 constructs have been produced to be used as tools for deciphering induction of pluripotency and differentiation in bovine embryonic derived cell lines. These have been transfected into the CT-1 line to determine how they regulate other key genes in this TE bovine cell line. Knock-down of endogenous gene expression using siRNA techniques has also been performed. Real-time PCR results are being collected and analyzed. Transcripts from an OCT4 pseudogene located on bovine chromosome 7 were detected in addition to transcript from the OCT4 gene. While full length Oct4 protein was detected by Western blot, the pseudogene mRNA does not appear to translated at a detectable level. A contiguous sequence has been reconstructed by RT-PCR which indicates that the cDNA is composed of a degenerate processed pseudogene of bOCT4 exons 1-4. Samples were collected from in vitro and in vivo produced bovine blastocysts, primary cell cultures established from ICM outgrowths, and from an established trophectoderm cell line (CT-1). The samples were processed and cDNA sequenced using the Illumina system. Data will be analyzed this year. Identified microRNA transcripts present in bull spermatozoa. Determined single nucleotide polymorphisms (SNPs) associated with bull fertility. Identified spermatozoa proteins associated with bull fertility. Contributed to better understanding of bovine genome in the Bovine Genome Sequencing and Annotation Consortium. Elucidated epigenetic regulators of early embryonic development, i.e., chromatin remodelers and DNA methyltransferases. Evaluated in vitro culture systems for porcine oocyte maturation, fertilization and embryo culture. Established an in vitro follicle culture system to identify the effect of excess insulin and IGF exposure on oocyte gene expression. Chronic in vitro exposure of follicles to insulin causes changes in the expression of NIMA-related kinase 2 (Nek2), Nek4 and TACC1. Similarly, insulin treatment of granulosal cell primary cultures induced alterations in Nek2, Nek4 and TCAA1 mRNA abundance. Determined the expression profile of Nek2, Nek4 and TCAA1 during the follicular and luteal phases of the estrous cycle in superovulated mice. Interestingly, Nek2 and Nek4 mRNA and protein expression seemed to be uncoupled with mRNA abundance peaking during the follicular phase and protein abundance peaking during the luteal phase. We have determined an important role for GnRH during embryogenesis, having a receptor-mediated effect on 1-cell embryos that occurs within the first 36 hours of embryo development. A specific antagonist of GnRH completely blocks embryonic development and acts via alteration of the cell cycle rather than apoptotic pathways. In addition, we have correlative data to suggest that GnRH binding to its receptor in early embryos activates the protein kinase C signaling pathway. Via biotin studies, we have accomplished the following: (i) established biotin effects on oocyte maturation in mice; (ii) identified genes that are up- or down-regulated in oocytes from biotin-deficient compared to biotin-normal mice; and (iii) examined intracellular signaling cascades and molecular mechanisms underlying biotin effects on oogenesis and subsequent development. Transgenic pigs were produced expressing a spermatogonial marker gene. mRNA and proteins were successfully extracted from both freshly ejaculated and frozen samples of stallion spermatozoa. A significant correlation between immunostaining for the fertility-related protein, SP22, was found with total motility and progressive motility for semen collected during the breeding season. Identified potential candidate mRNAs associated with FSH-induced initiation of GVBD in murine and bovine cumulus-oocyte complexes matured in vitro. Used short interfering RNA approaches to assess the functional roles of Nr4A1 and Egr1 mRNAs during FSH-induced oocyte maturation in cattle. Determined that the administration of FSH using 3 subcutaneous injections in combination with a progesterone-releasing device was an effective method for superovulation of Holstein cows. Identified that bovine AIRN RNA is expressed in cattle. Found expression of bovine AIRN RNA in fetal tissue at the post-implantation and peri-implantation stages but not in pre-implantation stages of development. The modification of histones is an epigenetic mechanism used to regulate gene expression. Abnormal modifications of histones have been proposed to contribute to the reduced development to term and low success rate of embryos following somatic cell nuclear transfer (scNT). The inability to reprogram epigenetic markers such as histone modifications found in the fibroblast donor cell line, especially in histones associated with establishing totipotency, could add to the reprogramming deficiencies associated with scNT. Two histone modifications associated with transcriptional activation; H3K4m3 and H4K16ac, and one histone modification associated with repressing transcription; H3K9m2, were considered with their association to three genes known to contribute to maintaining totipotency: NANOG, POU5F1, and SOX2. The uChIP protocol was performed by using antibodies specific for each histone modification, follwed by real time PCR (qPCR) analysis in order to compare bovine cumulus cells, scNT blastocysts, and in vitro fertilized (IVF) blastocysts. The gene POU5F1 was not highly associated with any of the histone modifications regardless of the treatment group. The Sox2 gene showed high levels of association with all three histone modifications in both IVF and scNT blastocysts, but very minimal association with the modifications in cumulus cells. The gene Nanog was highly associated with all three histone modifications in the scNT embryos, while only highly associated with the modifications H3K4m3 and H3K9m2 in IVF embryos. Transcription factors play a critical role in gene regulation and the characterization of these DNA binding proteins is crucial for elucidating some of the interacting proteins that assist in regulating transcription. The interaction of proteins with DNA corresponding to different portions of the promoter regions of the bovine Oct4 and Sox2 genes was evidenced by a mobility shift in the band migration compared to the control DNA. The shifted bands were excised from the 6% polyacrylamide gel and were subsequently submitted to mass spectrometry analysis for identification of candidate proteins. Putative proteins identified are involved in diverse cellular transcriptional processes including: transcriptional activation; chromatin modification, decondensation, and distribution; nuclear architecture including core histone 2a; constitutive and alternative splicing; DNA repair; regulation of RNA polymerase II; and DNA stabilization. Intracellular calcium (Ca2+i) release, a hallmark of oocyte activation, is the end result of a complex series of signal transduction pathways which have yet to be completely characterized in the bovine model. It is well known that src family kinases (SFK) activate Phospholipase C (PLC) which converts phosphatidyl inositol (4,5)-bisphosphate into diacylglycerol and 1,4,5-inositol trisphosphate (IP3) which is directly involved in releasing Ca2+i from the endoplasmic reticulum. A two prong approach was undertaken to identify the mechanisms involved in the signal transduction pathway resulting in bovine activation. The first approach utilized immunoblotting to identify the presence of endogenous PLC isoforms in total bovine oocyte lysate using primary antibodies directed against four distinct isozymes of PLC. Immunoblotting was performed according to standard laboratory protocols and confirmed the presence of the PLC isoforms ´1, ´3, ´4, ³1, ²3 and ²4. The second approach involved microinjecting primary antibodies of ten different PLC isoforms into in vitro matured bovine oocytes at a 1:100 dilution. Following microinjection, oocytes were fertilized and cultured in vitro according to standard laboratory procedures (Reed et al., 1996). Activation and development were assessed by recording cleavage at 48 hours post fertilization. Microinjection of several PLC-specific antibodies resulted in no effect on cleavage rates while others significantly blocked development. Determining the presence and involvement of these intracellular signaling molecules in bovine oocyte fertilization and activation will result in better activation protocols for nuclear transfer (NT). Deficiencies in the success rate of somatic cell nuclear transfer are widely held to be epigenetic in nature, and arise from the limited ability of a differentiated donor cell to erase epigenetic signatures required for nuclear reprogramming. Following bovine somatic cell nuclear transfer DNA methylation signatures of two genes necessary for pluripotency and self-renewal, namely Nanog and POU5F1 (Oct-4) more closely resemble that of somatic cells rather than in vitro fertilized embryos. A retained methylation signature following scNT likely leads to interaction with methyl binding domain proteins capable of binding to methylated promoter regions and acting to silence gene expression. Using a modified version of Electrophoretic Mobility Shift Assay (EMSA) targeted to the genes Nanog and POU4F1 (Oct-4) we identified DNA binding proteins that interact specifically with the methylated promoter regions. We observed protein binding specific to a methylated DNA template that differs from the binding proteins specific to the non-methylated DNA template. These findings indicate that the gene promoter region is acted upon by different DNA binding proteins depending on its methylation status, and that the retention of a hyper-methylation signature could lead to the premature down-regulation of the genes Nanog and POU5F1. The practical application of somatic cell nuclear transfer (SCNT) in animal agriculture and biomedical research is limited at the present time to technical inefficiencies that contribute to high rates of embryo and fetal loss. One of those inefficiencies that has yet to be optimized in the animal system is the enucleation procedure. In this study we evaluated various enucleation methods that might lead to increased efficiency of chromosome removal in the bovine oocyte. The results of the study are summarized as following. 1. Three-tenth M of sucrose and 0.6ug/ml demecolcine will induce high cytoplasmic protrusions containing chromosomes (75% and 90%, respectively). 2. Most cytoplasmic protrusions induced by sucrose treatment will last longer than 30 min; whereas, demecolcine induced protrusions will disappear within 30 min, particularly when oocytes are exposed to solutions containing 7.5ug/ml cytochalasin B (CB). 3. Enucleation of 100% can be achieved upon removing cytoplasmic protrusions with a micropipette in 0.3M sucrose and 7.5ug/ml CB. The survival rate of oocytes using this procedure, however, is negatively impacted (reduced to 75%), and we therefore consider this as a limiting factor in its application . 4. One hundred percent enucleation is achieved by removing induced cytoplasmic protrusions in the presence of demecolcine. Oocyte survival rate is high when enucleation is performed in CB-free solution with or without demecolcine. 5. The cleavage and blastocyst rates of cloned embryos from sucrose or demecolcine-treated oocytes are similar. 6. Oocyte microtubules are depolymerized and mostly depleted in demecolcine or demecolcine+CB treated oocytes, but remain nearly intact in sucrose-treated oocytes. The results of this study indicate that the enucleation of bovine oocytes in sucrose+CB or demecolcine+CB solutions are inefficient. CB does not appear to be required to enucleate bovine oocytes, and demecolcine-assisted enucleation is just as efficient when not exposing oocytes to CB. Functional characterization of importin ±8 specifically expressed in bovine oocytes and early embryos. Our previous results support an important functional role for importin ±8 in early embryogenesis. We have initiated new experiments to test the ability of importin ±8, relative to other importin ±s, to bind and facilitate nuclear transport of known oocyte-specific transcription factors important for oocyte and early embryonic development, and the functional requirement of importin ±8 for meiotic maturation, fertilization and specific developmental events characteristic of the maternal-to-embryonic transition. Through data mining, we have identified the cDNA sequences for all bovine importin ± genes. The predicted importin ±1, ±3, ±4, ±5, ±6 and ±7 proteins are 529 aa, 521 aa, 521 aa, 538 aa, 536 aa and 536 aa, respectively. All proteins are predicted to contain an IBB domain and 8 ARM repeats. The importin ±8 protein shares 53%, 45%, 45%, 41%, 40% and 40% sequence identity with importin ±1, ±3, ±4, ±5, ±6 and ±7, respectively. Phylogenetic analysis revealed that importin ±8 belongs to the first subfamily of the importin ± family. Expression of mRNA for importin ±1, ±3, ±4, ±5, ±6 and ±7 in bovine tissues was determined by RT-PCR. All 6 genes are ubiquitously expressed. We have cloned the coding regions of these genes as well as the importin ±8 gene in frame of the glutathione S-transferase (GST) gene in pGEX-4T-1 vector and successfully expressed the GST-importin fusion proteins. We have also obtained the bovine cDNA sequences for a number of oocyte-specific genes including Oct4 and Nobox. We are currently evaluating the interactions of all importin ± proteins with Oct4 and Nobox proteins by GST pull-down assay. Furthermore, we have validated a model system where, following siRNA injection, spontaneous germinal vesicle breakdown can be inhibited for 48 h of culture in vitro in the presence of S-roscovitine. This model system will be used for studies of effects of importin ±8 reduction during meiotic maturation, fertilization and initial cleavage divisions. Objective 2: Refine methods for production of genetically modified animals to improve livestock production efficiency We have identified all members of the importin ± gene family in cattle, prepared plasmid constructs, and validated methodologies for future experiments to elucidate the functional roles of importin ±8 during early events of embryonic development. Work is currently in progress to determine the time-course protocols to assess the efficiency of non-homologous and homologous recombination in primary cells and in conjunction with cell cycle manipulations. Data suggest the time course of the effect is extended in primary cells and cell viability is decreased, especially following treatment with Ku 70 siRNA, which can be partially rescued by siRNA depletion of P53 Investigated the feasibility of using frozen and immature oocytes in the generation of embryonic stem cells; studied genetic imprinting in naturally reproduced and cloned calves. For the first time in a domestic livestock species, porcine induced pluripotent stem cells were demonstrated to give rise to chimeric offspring demonstrating a true pluripotent state. These cells can undergo complex genetic manipulations to produce transgeneic animals that significantly improve livestock production. The present results suggest that there is no horizontal Tg transmission between T and C pigs due to rearing or mating. This work provides a critical step toward providing rigorous scientific data for risk assessment of transgenic livestock. ADSC and BMSC can be successfully differentiated into adipogenic and osteogenic lineages and maintain viability in an alginate hydrogel over time. Our conclusion, is that alginate is a viable scaffold material for the differentiation of mesenchymal stem cells for tissue engineering applications. The combination of microarray data and pairwise analysis uncovered novel and high reliable ICG for qPCR normalization in adult porcine stem cells induced into adipogenic and osteogenic lineages. Results suggest both ADSC and BMSC can differentiate towards the adipogenic lineage but with quantitatively different gene expression patterns. The use of multi-factorial directed differentiation using high-speed robotic systems will enable the examination of large matrices of culture and differentiation conditions for stem cells. Using the automated microscale system in large factorial experiments allows analysis of the basic mechanisms underlying stem cell development in vitro, and ultimately in vivo. We have identified genes whose products might be playing a role in epigenetic control of molecular reprogramming of gene expression in cloned bovine embryos. Constructed lentiviral vectors containing a constitutively active promoter fused to selected siRNA designed to reduce GnRHR II mRNA levels in porcine cells and tissues. We have developed a successful procedure for cryopreservation of porcine embryos using a microdroplet procedure. In addition, we have established divergent survival rates for cryopreserved embryos from Chinese Meishan and white crossbred lines of swine. This difference in survival rate occurs within the first 24 hours following thawing. Transgene copy number following transfection with alternative methods has been determined. It was determined that electroporation results in an average of 2-3 copies of the transgene while liposome-mediated transfection yields predominantly single copy insertions. Isolation and culture procedures for adipose derived stem cells from cattle and pigs have been established. These stem cell populations have been characterized for cell cycle characteristics, chromosomal stability in culture and epigenetic modifications. Bovine and eland adipose derived stem cells have been utilized as donor cells in somatic cell nuclear transfer procedures. Expression of pluripotency genes has been studied in bovine fetal fibroblasts and adipose derived stem cells.

Refereed articles Caldwell, J. D., K. P. Coffey, W. K. Coblentz, J. A. Jennings, D. S. Hubbell, III, D. L. Kreider, M. L. Looper, and C. F. Rosenkrans, Jr. 2009. Performance by fall-calving cows grazing tall fescue pastures with different proportions stockpiled. Forage and Grazinglands doi:10.1094/FG-2009-0312-01-RS. Looper, M. L., G. E. Aiken, and C. F. Rosenkrans, Jr. 2009. Management strategies for cattle grazing toxic tall fescue during summer months. Forage and Grazinglands doi:10.1094/FG-2009-1102-03-RV. Looper, M. L., T. S. Edrington, T. R. Callaway, and C. F. Rosenkrans, Jr. 2009. Fate of Escherichia coli O157:H7 and Salmonella from contaminated manure slurry applied to soil surrounding tall fescue. Lett. Appl. Micro. 48:513-516. Looper, M. L., T. S. Edrington, and C.F. Rosenkrans, Jr. 2009. Influence of body condition and forage type on prevalence of Escherichia coli O157:H7 and Salmonella in grazing beef cows. Lett. Appl. Micro. 49:361-365. Looper ML, Rorie RW, Person CN, Lester TD, Hallford DM, Aiken GE, Roberts CA, Rottinghaus GE, Rosenkrans CF Jr. 2009. Influence of toxic endophyte-infected fescue on sperm characteristics and endocrine factors of yearling Brahman-influenced bulls. J Anim Sci. 87:1184-1191. Wang, H., M.L. Looper, Z.B. Johnson, R.W. Rorie and C.F. Rosenkrans. 2009. Involvement of signaling pathways in bovine sperm motility and effect of ergot alkaloids. In Vitro Cell Dev Biol Anim. 45:483-489. Looper, M. L., S. G. Black, S. T. Reiter, R. Okimoto, Z. B. Johnson, M. A. Brown, and C. F. Rosenkrans, Jr. 2010. Identification of polymorphisms in the enhancer region of the bovine prolactin gene and association with profitability traits of beef cattle. Prof. Anim. Sci. (in press). Rosenkrans, C., Jr., A. Banks, S. Reiter, M. Looper. 2010. Calving rates of Brahman-influenced cows are associated with heat shock protein 70 genetic polymorphisms. Anim. Repro. Sci. (in press). Berger, T., B. Nitta, Y. Ducolomb, and M. Betancourt. 2009. Interaction of potential porcine sperm ligands with the oocyte plasma membrane. Reprod Domest Anim. [Epub ahead of print] Berger, T., and B. M. Roberts. 2009. Reduced immunolabelling of a porcine oocyte membrane protein reflects reduced fertilizability of porcine oocytes following elevated ambient temperature. Reprod Domest Anim 44: 260-265. Bertolini, L.R., M. Bertolini, E.A. Maga, E.A., K.R. Madden and J.D. Murray. 2009. Increased gene targeting in Ku70 and Xrcc4 transiently deficient human somatic cells. Mol. Biotech. 41:106-114. Corbin, C. J. et al. 2009. Porcine hypothalamic aromatase cytochrome p450: Isoform characterization, sex-dependent activity, regional expression, and regulation by enzyme inhibition in neonatal boars. Biol Reprod 81: 388-395. Ahola, J.K., G.E. Seidel, Jr. and J.C. Whittier. 2009. Use of gonadotropin-releasing hormone at fixed-time artificial insemination at eighty or ninety-seven hours post prostaglandin F2± in beef cows administered the long-term melengestrol acetate Select Synch. Prof Anim Sci 25:256-261. Barcelo-Fimbres, M., Z. Brink and G.E. Seidel, Jr. 2009. Effects of phenazine ethosulphate during culture of bovine embryos on pregnancy rate, prenatal and postnatal development after embryo transfer. Theriogenology 71:355-368. Barfield, J.P., P.M. McCue, E.L. Squires and G.E. Seidel, Jr. 2009. Effect of dehydration prior to cryopreservation of large equine embryos. Cryobiology 59:36-41. Campos-Chillon, L.F., T.K. Suh, M. Barcelo-Fimbres, G.E. Seidel, Jr. and E.M. Carnevale. 2009. Vitrification of early-stage bovine and equine embryos. Theriogenology 70:349-354. Hyland, A., G.E. Seidel, Jr., R.M. Enns, R.K. Peel and J.C. Whittier. 2009. Intervals of five or seven days between controlled internal drug-release insertion, gonadotropin-releasing hormone, and prostaglandin F2± injections: Effects on pregnancy rate and follicular size. Prof. Anim. Sci. 25:150-154. Mahmoud, K.G., T.H. Scholkamy, Y.F. Ahmed, G.E. Seidel, Jr. and M.F. Nawito. 2009. Effect of different combinations of cryoprotectants on in vitro maturation of immature buffalo (Bubalus bubalis) oocytes vitrified by straw and open-pulled straw methods. Reprod. Dom. Anim. PMID 19090828. Purcell, S.H., J.D. Cantlon, C.D. Wright, L.E. Henkes, G.E. Seidel, Jr. and R.V. Anthony. 2009. The involvement of proline-rich 15 in early conceptus development in sheep. Biol. Reprod. 81:1112-1121. Roberts, R.M., G.W. Smith, F.W. Bazer, J. Cibelli, G.E. Seidel, Jr., D.E. Bauman, L.P. Reynolds and J.J. Ireland. 2009. Farm animal research in crisis. Science 324:468-469. Seidel, G.E., Jr. 2009. ASAS Centennial Paper: Future research in physiology and endocrinology. J. Anim. Sci. 87:384-389. Mansouri-Attia N, Sandra O, Aubert J, Degrelle S, Everts RE, Giraud-Delville C, Heyman Y, Galio L, Hue I, Yang X, Tian XC, Lewin HA, Renard JP. 2009. Endometrium as an early sensor of in vitro embryo manipulation technologies. Proc Natl Acad Sci U S A. 2009 Apr 7;106(14):5687-92. Epub 2009 Mar 18. Mansouri-Attia N, Aubert J, Reinaud P, Giraud-Delville C, Taghouti G, Galio L, Everts RE, Degrelle S, Richard C, Hue I, Yang X, Tian XC, Lewin HA, Renard JP, Sandra O. 2009. Gene expression profiles of bovine caruncular and intercaruncular endometrium at implantation. Physiol Genomics. 2009 Sep 9;39(1):14-27. Epub 2009 Jul 21. Curchoe CL, Zhang S, Yang L, Page R, Tian XC. 2009. Hypomethylation trends in the intergenic region of the imprinted IGF2 and H19 genes in cloned cattle. Anim Reprod Sci. 2009 Dec;116(3-4):213-25. Epub 2009 Feb 11. Suteevun-Phermthai T, Curchoe CL, Evans AC, Boland E, Rizos D, Fair T, Duffy P, Sung LY, Du F, Chaubal S, Xu J, Wechayant T, Yang X, Lonergan P, Parnpai R, Tian XC. Allelic switching of the imprinted IGF2R gene in cloned bovine fetuses and calves. Anim Reprod Sci. 2009 Nov;116(1-2):19-27. Epub 2009 Jan 20. Smith SL, Everts RE, Sung LY, Du F, Page RL, Henderson B, Rodriguez-Zas SL, Nedambale TL, Renard JP, Lewin HA, Yang X, Tian XC. 2009. Gene expression profiling of single bovine embryos uncovers significant effects of in vitro maturation, fertilization and culture. Mol Reprod Dev. 2009 Jan;76(1):38-47. Yang X, Guo XM, Wang CY, and Tian XC. 2009. Protocols for Large-Scale Derivation of Cardiomyocytes from Embryonic Stem Cells Chapter 22. In: Essentials of Stem cells. Elsevier, Lanza R (ed). (in press). Sung LY, Amano T, Smith SL, Tian XC and Yang X. 2009. Somatic Cell Nuclear Transfer and Derivation of Embryonic Stem Cells. Chapter 1. In: Methods in Stem Cell Medicine and Bioengineering (in press). Tian XC, Park J, Bruno R, French R, Jiang L, Prather RS. 2009. Altered gene expression in cloned piglets. Reprod Fertil Dev. 2009;21(1):60-6. Marjani SL, Le Bourhis D, Vignon X, Heyman Y, Everts RE, Rodriguez-Zas SL, Lewin HA, Renard JP, Yang X, Tian XC. 2009. Embryonic gene expression profiling using microarray analysis. Reprod Fertil Dev. 2009;21(1):22-30. Chang CC, Sung LY, Amano T, Tian XC, Yang X, Nagy ZP. Nuclear transfer and oocyte cryopreservation. Reprod Fertil Dev. 2009;21(1):37-44. West, F. D. et al. 2008. Enrichment and Differentiation of Human Germ-Like Cells Mediated by Feeder Cells and Basic Fibroblast Growth Factor Signaling. Stem Cells. 11: 2768-76. West, F. D. et al. 2010. Kit ligand and bone morphogenetic protein signaling enhances human embryonic stem cell to germ-like cell differentiation. Hum Reprod 25: 168-178. West, F. D. et al. Porcine Induced Pluripotent Stem Cells Produce Chimeric Offspring. Stem Cells and Dev (In Review; 12/01/09). West, F. D. et al. Isolation and Expansion of Meiotic Competent Clonal Germ Cell Lines Derived from Male Human Embryonic Stem Cells. Stem Cells and Dev (In Preparation; 01/06/09). Mônaco, E., Lima, A.S., Bionaz, M, Maki, A., Wilson, S.M., Hurley, W.L. Wheeler, M.B. 2009. Morphological and Transcriptomic Comparison of Adipose and Bone Marrow Derived Porcine Stem Cells. Open Tiss. Eng. Regen. Med. J. 2:20-33. Bleck, G.T., Wheeler, M.B., Hansen, L.B., Chester-Jones, H.,, Miller, D.J. 2009. Lactose Synthase Components in Milk: Concentrations of a-Lactalbumin and b1,4-Galactosyltransferase in Milk of Cows from Several Breeds at Various Stages of Lactation. Reprod. Dom. Anim 44:241247. Yuan, Y., Krisher, R.L. (2009) Effect of ammonium during in vitro maturation on porcine oocyte nuclear maturation and subsequent embryonic development. Animal Reproduction Science 117:302-307. doi:10.1016/j.anireprosci.2009.05.012. Paczkowski, M., Krisher R.L. (2010) Aberrant Protein Expression is Associated with Decreased Developmental Potential in Porcine Oocytes. Molecular Reproduction and Development 77:51-58. Published Online: 2 Sep 2009; DOI 10.1002/mrd.21102. Krisher, R.L., Wheeler, M.B. (2010). Towards use of microfluidics for individual embryo culture. Reproduction, Fertility and Development 22:32-39. Giraldo, A.M., M.N. Purpera, T.D. Vaught, D.L. Ayares, J W. Lynn, R. A. Godke and K R. Bondioli. 2009. Inhibition of DNA methyltransferase 1 expression in bovine fibroblast cells used for nuclear transfer. Reprod. Fertil. Develop. 21:785-795. Purpera, N.M., A.M. Giraldo, C. B. Ballard, D. Hylan, R. A. Godke and K. R. Bondioli. 2009. Effects of culture medium and protein supplementation on mRNA expression of in vitro produced bovine embryos. Molec. Reprod. Dev. 76(8):783-793. Hylan, D., A. M. Giraldo, J. A. Carter, G.T. Gentry Jr., K. R. Bondioli and R.A. Godke. 2009. Sex ratio of bovine embryos and calves originating from the left and right ovaries. Biol Reprod. 81:933-938. Owiny, O.D., D.M. Barry, M. Agaba and R.A. Godke. 2009. In vitro production of cattle x buffalo hybrid embryos using cattle oocytes and African buffalo (Syncerus caffer caffer) epididymal sperm. Theriogenology 71:884-894. Alapathi, R., M. Stout, J. Saenz, G. T. Gentry Jr., R.A. Godke and R.V. Devireddy. 2009. Ejaculated and epidydimal bovine sperm exhibit equivalent freezing response. Cryobiology 59: 164-170. Wirtu, G., C.E. Pope, D.L Paccamonti, R.A. Godke, B.L Dresser. 2009. Ultrasound-guided retrieval and in vitro maturation of eland (Taurotragus oryx) and bongo (Tragelaphus eurycerus isaaci) antelope oocytes. Anim. Reprod. Sci. 111:160-172. Hylan, D., A. M. Giraldo, K. R. Bondioli, R. A. Godke. 2009. Distribution of sexes within the left and right uterine horns of beef cattle. Theriogenology (In press) Godke, R.A., G.T. Gentry Jr. and K. R. Bondioli. New biotechnologies for the farm animal industry. So. Assn. Agri. Scientists. (Refereed) (In press) Pant, D., and C. L. Keefer. 2009. Expression of pluripotency-related genes during bovine inner cell mass explant culture. Cloning Stem Cells 11: 355-365. Rodriguez-Osorio, N., H. Wang, J. Rupinski, S. M. Bridges and E. Memili. Comparative functional genomics of mammalian DNA methylation. In press. Reproductive Biomedicine online. Bovine Genome Sequencing and Annotation Consortium including Memili, E., and others. 2009. The genome sequence of Taurine Cattle: A window to ruminant biology and evolution. Science 324(5926):522-528. Rodriguez-Osorio, N., Z. Wang, P. Kasinathan, G. P. Page, J. M. Robl and E. Memili. 2009. Transcriptional reprogramming of gene expression in bovine somatic cell chromatin transfer embryos. BMC Genomics 10:190. Uzun, A., N. Rodriguez-Osorio, H. Wang, A. Kaya, J.J. Parrish, V. Ilyin and E. Memili. 2009. Comparative Functional Genomics of Chromatin Remodeling Proteins HMGN3a and SMARCAL1 During Early Mammalian Embryogenesis. BMC Genomics 10:183. Feugang, J. M., A. Kaya, G.P. Page, L. Chen, T. Mehta, K. Hirani, L. Nazareth, R. A. Gibbs and E. Memili. 2009. Two stage genome wide association study identifies Integrin beta five as having a potential role in bull fertility. BMC Genomics 10: 176. Wang, H., J. M. Feugang, N. Rodriguez-Osorio, S. Jung, K. Garrison, C. Wolgemuth, L. Greer, E. Memili. 2009. Effects of culture media and inhibitors on biology of porcine embryonic development in vitro. Livestock Science 121: 102-107. Feugang, J. M, O. D. Camargo-Rodriguez and E. Memili 2009. Culture Systems for Bovine Embryos. Livestock Science 121:141-149. Cherrington, B.D., T.A. Farmerie, B.E. Bass, R.A. Cederberg, C.M. Clay, and B.R. White. 2009. Emergence of a new regulatory element in the proximal promoter of the gonadotropin releasing hormone receptor in Chinese Meishan swine. Biol. Reprod. (Provisionally Accepted). Montagner, M.M, A.R. Cropp, J.J. Swanson, R.A. Cederberg, P.B.D. Goncalves, and B.R. White. 2009. Role of gonadotropin-releasing hormone on mouse preimplantation embryonic development. Mol. Reprod. Dev. (Submitted). Montagner, M.M, P.B.D. Goncalves, G.A. Mills, R.K. Christenson, and B.R. White. 2009. Divergent survival of Meishan and white crossbred porcine embryos following vitrification using a microdroplet method. Reprod. Fert. Dev. (Submitted). Silva, C., J.R. Wood, L. Salvador, Z. Zhang, I. Kostetskii, C.J. Williams, and J.F. Strauss III 2009. Expression profile of male germ cell-associated genes in mouse embryonic stem cell cultures treated with all-trans retinoic acid and testosterone. Mol. Reprod. Dev. 76:11-21. Zhang, Z., R. Jaimez, X. Shen, D. Gude, H. Tang, J.R. Wood, E. Goldberg, and J.F. Strauss III. 2009. Autoregulation of SPAG16 expression: A single gene encoding an axoneme structural protein and a transcription factor that activates the axoneme protein promoter. Nucleic Acids Res. (Submitted). Farin CE, Farmer WT, Farin PW. 2010. Pregnancy recognition and abnormal offspring syndrome. Reprod Fertil Dev 22:75-87. Wrench N, Pinto CRF, Klinefelter GR, Dix DJ, Flowers WL, Farin CE. (2010). Effect of season on fresh and cryopreserved stallion semen. Anim Reprod Sci (accepted). Farmer WT, Farin PW, Piedrahita JA, Bischoff SR, Farin CE. (2010). Detection of antisense of insulin-like growth factor-2 receptor RNA non-coding (AIRN) in cattle. Reprod Fertil Dev (in revision). Farin CE, Alexander JE, Farin PW. (2010). Expression of messenger RNAs for insulin-like growth factors and their receptors in bovine fetuses at early gestation from embryos produced in vivo or in vitro. Theriogenology (in revision). Li, G.P., K.L. White , K.I. Aston , T.D. Bunch, B.A. Hicks, Y. Liu , and B.R. Sessions. 2009. Colcemid-treatment of heifer oocytes enhances nuclear transfer embryonic development, establishment of pregnancy and development to term. Mol. Reprod. Dev. 76:620-628. Aston, K.I., G.P. Li, B.A. Hicks, B.R. Sessions, A.P. Davis, Q.A. Winger, L.F. Rickords, and K.L. White. 2009. Global gene expression analysis of bovine somatic cell nuclear transfer blastocysts and cotyledons. Mol Reprod Dev. 76:471-82. Tejomurtula, J., Lee, K.B., Tripurani, S.K., Smith, G.W. and Yao, J. 2009. Role of Importin ±8, a new member of the importin ± family of nuclear transport proteins, in early embryonic development in cattle. Biology of Reproduction. 81: 333342. Books, non-refereed book chapters, proceedings, instructional media, theses/dissertations Delgado Callisaya, P.A. 2009. Factors influencing survival of bovine spermatozoa during storage and related to the ratio of X- to Y-chromosome bearing spermatozoa. M.S. Thesis, University of Arkansas, Fayetteville Seidel, G.E., Jr. 2009. Sperm sexing technology  the transition to commercial application. Theriogenology 71:1-3. Seidel, G.E., Jr. 2009. Artificial insemination of cattle with sexed semen. Proc. Meeting on Milk and Meat Production in Warm Climates, pp. 43-51. Wall, R.J., G. Labile, E. Maga, G.E. Seidel, Jr. and B. Whitelaw. 2009. Animal productivity and genetic diversity: Cloned and transgenic animals. In: Animal Agricultures Future through Biotechnology, Part 8, Council for Agricultural Service and Technology, Ames, IA, 16 pp. West, F. D. Differentation of Male Human Embryonic Stem Cells into Germ-Like Cells: A Developmental Model and an Assisted Reproductive Technology. Roberts, J. 2009. Title: Effect Priming with Melegestrol Acrtate (MGA) on Subsquent Fertility in Beef Heifers. MS Thesis. Louisiana State University. Baton Rouge, LA. Spring (May) 2009. Pennington, P 2009. Title: Characterization of the Common Eland (Taurotragus oryx) Estrous Cycle. MS Thesis Louisiana State University. Baton Rouge, LA. Summer 2009 Willaims, K. 2009. Title: Characterization of Adult Porcine Adipose Stem Cells PhD Dissertation. Louisiana State University. Baton Rouge, LA. Summer 2009. (K. Bondioli and R. Godke Joint Advisors). Picou, A. 2009. Title: Isolation and Characterization of Bovine Adipose Derived Somatic Stem Cells for the use in Nuclear Transfer. MS Thesis. Louisiana State University, Baton Rouge, LA. Summer 2009. Wilson, J. 2009. Title: Determining Gene Copy Number in Transfected Caprine Fibroblasts. MS Thesis. Louisiana State University, Baton Rouge, LA. Summer 2009. Denniston, R. S., S. Michelet, K.R. Bondioli and R. A. Godke. Principles of Embryo Cryopreservation. In: Cryopreservation in Aquatic Species. Tiersch, T. R. and P. M. Mazik, Editors. World Aquaculture Society, Baton Rouge, Louisiana. (In press). Chandler J.E. and R. A. Godke. Cryopreservation of Cattle Sperm. In: Cryopreservation in Aquatic Species. Tiersch, T. R. and P. M. Mazik, Editors. World Aquaculture Society, Baton Rouge, Louisiana. (In press). Brauer, V.M. 2009. Transcriptional regulation of the porcine type II GnRH receptor gene. MS Thesis. University of Nebraska, Lincoln, NE. Abstracts Barcelo-Fimbres, M. and G.E. Seidel, Jr. 2009. Effects of lipolytic agents forskolin, epinephrine and caffeine on embryonic development and lipid content of bovine embryos produced in vitro. Reprod. Fertil. Develop. 21:154-155. Preis, K.A., G.E. Seidel, Jr. and D.K. Gardner. 2009. Effect of in vitro maturation medium on subsequent quantity and quality of developing embryos. Proc. American Society of Reproduction Medicine. In Press. Barfield, J.L., R. Sanchez, E.L. Squires and G.E. Seidel, Jr. 2009. Vitrification and conventional cryopreservation of equine embryos. Reprod. Fertil. Develop. 21:130. Amorim, L.E., C.A.A. Torres, E.A.M. Amorim, J.D. Guimaraes, J.F. Fonseca, L.G.B. Sigueira and G.E. Seidel, Jr. 2009. Effect of dietary urea on embryonic viability and development of Toggenburg goats. Reprod. Fertil. Develop. 21:157. Jiang L, SL Marjani, M Bertolini, H A Lewin, GB Anderson, X Yang, and XC Tian. 2010. Indistinguishable Transcriptional Profiles Between In Vivo- and In Vitro-Produced Bovine Fetuses (presented at the 36th annual meeting of IETS, Cordoba, Argentina, Jan 9-12, 2010) Chang CC, Lin CJ, Friedman J, Kort HI, Tian XC, Nagy ZP. The difference of spindle recovery during oocyte cryopreservation: vitrification vs. slow freezing. Fertil Steril 2009;92(3S):S86. Oral presentation (Atlanta, Georgia, USA; American Society for Reproductive Medicine in 2009 annual meeting. Yang, X, Amano T, Ma Y, He Z, Amano M, Lin C-J, Treaster S, Dym M, Tian XC. 2009. Genetically corrected ES cells derived from cloned embryos of infertile mice  a potential application of therapeutic cloning. Presented at the 6th Annual meeting of the Asian Reproductive Biotechnology Society, Siem Reap, Cambodia, Nov 16-20, 2009. p. 3. Franklin West, Kurinji Pandiyan, Kelly Robbins and Steve Stice. Differentiation of Primordial Germ Like Cells From Human Embryonic Stem Cells. Society for the Study of Reproduction; 2009 Sept: Pittsburgh, (PA). F.D. West, M.S. Natrajan, M.I. Roche-Rios, M.A. Bedell and S.L. Stice. Bone Morphogenetic Protein and KIT Ligand Signaling Enhance Human Embryonic Stem Cell Differentiation into Early Sperm Cells. Georgia Life Sciences Summit; 2008 Sept: Atlanta, (GA). Franklin West, Kurinji Pandiyan, Kelly Robbins and Steve Stice. Enrichment and Differentiation of Germ-Like Cells from Human Embryonic Stem Cells. Georgia Stem Cell Initiative; 2008 May: Athens, (GA). Recipient of 3rd place in poster competition. Franklin West, Kurinji Pandiyan, Kelly Robbins and Steve Stice. Enrichment of Germ-Like Cells in Human Embryonic Stem Cells Cultures. GTECH Industry Partners Symposium; 2007 Oct: Atlanta, (GA). F.D. West, K. Pandiyan, K. Robbins and S. L. Stice. Human Embryonic Stem Cells Differentiate into Sperm and Eggs Precursor Cells. Georgia Life Sciences Summit; 2007 Oct: Atlanta, (GA). Franklin West, Kurinji Pandiyan, Kelly Robbins and Steve Stice. Enrichment of Germ-Like Cells in Human Embryonic Stem Cell Cultures. Society for the Study of Reproduction 40th Annual Meeting; 2007 Jul 21-25: San Antonio, (TX). West F., Pandiyan K., Robbins K. R., and Stice S. L. 2006. Differentiation of Primordial Germ Like Cells from Human Embryonic Stem Cells. The University of Georgia Biomedical and Health Science Institute 2006 Annual Retreat; 2006 Sept 22; Athens (GA). Bionaz, M., Monaco, E., Lima, A.S., Lane, S.J., Kim, D., Hurley, W.L., Wheeler, M.B. 2009. Internal control genes for quantitative PCR of porcine mesenchymal stem cells during adipogenic and osteogenic differentiation in vitro. Reproduction, Fertility and Development 21:188-189. Kim, D., Maki, A.J., Kong, H-J., Monaco, E., Bionaz, M., Hurley, W.L., Wheeler, M.B. 2009. Multilineage potential of porcine bone marrow and adipose-derived mesenchymal stem cells in 3-D alginate hydrogels. Reproduction, Fertility and Development 21:237. Monaco, E., Lima, A.S., Wilson, S.M, Lane, S.J., Bionaz, M., Hurley, W.L., Wheeler, M.B. 2009. Adipogenic differentiation in vitro of porcine adult mesenchymal stem cells. Reproduction, Fertility and Development 21:238. Wheeler, M.B., Hurley, W.L., Lane, S.J., Mosley, J., Bressner, G.E., Monaco, E., Wilson, S.M. 2009. Risk analysis of alpha-lactalbumin transgene transfer to non-transgenic control animals during rearing and breeding. Reproduction, Fertility and Development 21:252-253. M. Paczkowski, J. Fleming-Waddell, C.A. Bidwell, R.L. Krisher. (2009) Maternal Age Alters Fetal and Placental Development and Expression of Methylated Genes. Reprod Fertil Dev. 21(1):194 (abstr. 191). Silva, E., Krisher, R. (2009) Leptin and Glucose Influence Porcine Nuclear Maturation. Reprod Fertil Dev. 21(1):226 (abstr. 256). Yuan, Y., R.L. Krisher. 2009. Glutaredoxin pathway genes are differentially expressed in mature porcine oocytes with varying developmental potentials. Biology of Reproduction 81 (Suppl. 1): 370. Ohlweiler, L.U., Mezzalira, J.C., Monaco, E., Mezzalira, A., Bertolini, M., Wilson, S.M., Ringwelski, J., Krisher, R.L., Rund, L., Wheeler, M.B. (2010) Pregnancy outcome after oviductal transfer of zona free 1 cell stage porcine embryos produced by hand made cloning. Reproduction Fertility and Development 22(1):194 (abst. 72). Mezzalira, J.C., Ohlweiler, L.U., Massie, A., Monaco, E., Silva, E.P., Yuan, Y., Mezzalira, A., Bertolini, M, Krisher, R.L., Wheeler, M.B. (2010) Effects of cell type, pre-activation protocol and culture conditions on development of porcine hand made cloned embryos. Reproduction Fertility and Development 22(1):193 (abst. 69). Gentry Jr., G.T., J.A. Pitchford, M. Chiasson, L.R. Gentry, K.R. Bondioli, D.L. Thompson and R.A. Godke. 2009. Effect of circulating leptin levels at synchronization on subsequent fixed-timed artificial insemination pregnancy rates in crossbred beef heifers. Reprod. Fertil. Develop. 21:104. Guerrero, C.A., G.T. Gentry, J. Saenz, K.R. Bondioli, and R.A. Godke. 2009. Birth of calves after artificial insemination with cryopreserved bovine caudal epididymal spermatozoa harvested from a postmortem bull. Reprod. Fertil. Develop. 21:105. Saenz. J.R., C. Dumas, B.L. Dresser, M.C. Gómez, R.A. Godke and C.E. Pope. 2009. Survival and in vitro functionality of cat epididymal spermatozoa following cryopreservation in extenders with or without egg yolk. Reprod. Fertil. Develop. 21:138. Kish, S.L., J.A. Wilson, A.A. Picou, J.W. Lynn, R.A. Godke and K.R. Bondioli. 2009. Streptolysn O and permeabilized and mitotic extract treated fetal goat fibroblast for use in nuclear transfer. Reprod. Fertil. Develop. 21:119 Picou, A.A., R.A. Maclean, B.L. Dresser, R.G. Godke and K.R. Bondioli. 2009. Isolation and characterization of bovine adipose-derived somatic stem cells. Reprod. Fertil. Develop. 21:239 Pennington, P.M., .L.R. Gentry, C.E. Pope, R.A. Maclean. D.L. Paccamonti, B.L. Dresser, K.R. Bondioli, R.A. Godke and G. Wirtu. 2009. Characterization of the common eland (Taurotragus oryx) estrous cycle. Reprod. Fertil. Develop. 21:181 Stout, M.A,, J.R. Saenz, G.T. Gentry, S.P. Leibo, K.R. Bondioli and R.A. Godke. Comparison of post-thaw values of epididymal sperm from White-tail deer that was frozen using glycerol or DMSO as the cryoprotectant. Reprod. Fertil. Develop. 21:183. Williams, K.J., R.A. Godke and K. Bondioli. 2009. Characterization of porcine adipose tissue-derived adult stem cell surface proteins. Reprod. Fertil. Develop. 21:241 Wirtu, G., P.M. Pennington, C.E. Pope, R.A. MacLean, J. Mercado, J. Galiguis, D.L. Paccamonti, R.A. Godke and B.L. Dresser. 2009. Behavioral aspects of estrus in the common eland antelope, Taurotragus ory). Reprod. Fertil. Develop. Reprod. Fertil. Develop. 21:183 Carwell, D, J.A. Pitchford, G. Gentry Jr., H. Blackburn, K.R. Bondioli and R.A. Godke. Beef cattle pregnancy rates following insemination with aged frozen Angus semen. Reprod. Fertil. Develop. 22: (In press). Saenz, J.R., C. Dumasb, B. L. Dresser, M.C. Gómez R.A. Godke, C.E. Pope. Effect of egg yolk concentration in test buffered extender on survival and in vitro functionality of domestic cat epididymal spermatozoa following cryopreservation. Reprod. Fertil. Develop. 22: (In press). Pennington, P.M.,C. E. Pope, R. A. MacLean, B. L. Dresser, K. R. Bondioli, R. A. Godke, and G. Wirtu. Cortisol levels in the common eland (Taurotragus oryx) during intensive and nonintensive handling periods. Reprod. Fertil. Develop. 22: (In press). Chiasson, M.K., J.A. Carter, K.R. Bondioli, R.A. Godke and G.T. Gentry. Laser-assisted zona pellucida hatching in frozen-thawed bovine embryos. Reprod. Fertil. Develop. 22: (In press). Lambe J., W. Forbes, B.M. Olcott, D E. Sanders, R A. Godke and G.T. Gentry. Effect of GnRH on fixed-tmed artificial insemination pregnancy rates of White-tail deer. Reprod. Fertil. Develop. 22: (In press). Schiffmacher, A., and C.L. Keefer. 2009. Deciphering induction of pluripotency in a bovine model. Biology Reproduction 81(S1): abstr. 656. Robertson, L. R., J. M. Feugang, N. Rodriguez-Osorio, A. Kaya and E. Memili 2009. MicroRNAs of Bull Spermatozoa. Annual Conference of the International Embryo Transfer Society (IETS) in San Diego, CA, January 2009. Journal of Reproduction, Fertility and Development. Brauer, V.M., J.R. Wiarda, R.A. Cederberg, and B.R. White. 2009. Transcriptional regulation of the porcine gonadotropin releasing hormone II receptor gene. Biol. Reprod. 81(Supplement 1):352. Cederberg, R.A., V.M. Brauer, J.G. Kerl, J.R. Wiarda, and B.R. White. 2009. Characterization of the porcine Type II GnRH receptor gene. Biol. Reprod. 81(Supplement 1):371. Lee, C., R.A. Cederberg, and B.R. White. 2009. Glucocorticoid responsiveness of the porcine GnRH receptor (GnRHR) gene is conferred by an element(s) located between -290/-270 bp of proximal promoter. Biol. Reprod. 81(Supplement 1):161. Hicks JE, Farin, CE. 2009. Candidate mRNAs regulating meiotic resumption in bovine cumulus-oocyte complexes. Reprod Fertil Dev. 21:221 (abst #247). Farin PW, Dowdall KM, Hicks JE, Farin CE, Whisnant CS. 2009. Subcutaneous administration of follicle stimulating hormone for superovulation of Holstein cows. Reprod Fertil Dev. 21:243-244 (abst #293). Miscellaneous publications (semi-technical/lay publications) Ata, M.A., K.P. Coffey, J.D. Caldwell, A. N. Young, D. Philipp, E. Kegley, G.F. Erf, D.S. Hubbell, III, and C.F. Rosenkrans, Jr. 2009. Immune function responses of spring-born calves weaned from wild-type or novel-endophyte infected tall fescue. AR Agr. Exp. Sta. Research Series 574:60-62. Banks, A., M. Looper, S. Reiter, L. Starkey, and C. Rosenkrans, Jr. 2009. Associations between heat shock protein 70 genetic polymorphisms and calving rates of Brahman-influenced cows. AR Agr. Exp. Sta. Research Series 574:10-13. Caldwell, J., K. Coffey, D. Kreider, D. Philipp, D. Hubbell, III, J. Tucker, A. Young, M. Looper, M. Popp, M. Savin, J. Jennings, and C. Rosenkrans, Jr . 2009. Post-weaning performance by spring and fall-born calves weaned from full access, limited access, or no access to wild-type endophyte-infected tall fescue pastures  year 1. AR Agr. Exp. Sta. Research Series 574:49-51. Caldwell, J., K. Coffey, D. Kreider, D. Philipp, D. Hubbell, III, J. Tucker, A. Young, M. Looper, M. Popp, M. Savin, J. Jennings, and C. Rosenkrans, Jr. 2009 Performance by spring and fall-calving cows grazing with full access, limited access,or no access to wild-type endophtye-infected fescue  2 year summary. AR Agr. Exp. Sta. Research Series 574:52-54. P. A. Delgado, P.A., T.D. Lester and R.W. Rorie. 209. Effect of a Low-Sodium, Choline-Based Diluent on Viability of Bovine Sperm Stored at Refrigerator Temperatures. Ark Agri. Exp. Sta., Res. Series 574:77- 79. Larson, M., M. Sales, S. Reiter, H. Brown, Jr., M. Brown, M. Looper, and C. Rosenkrans, Jr. 2009. Effects of forage type and CYP3A28 genotype on beef cow milk traits. AR Agr. Exp. Sta. Research Series 574:18-21. Looper, M. L., S. T. Reiter, D. M. Hallford, and C. F. Rosenkrans, Jr. 2009. Influence of body condition and forage type on endocrine factors and calving rate of postpartum beef cows. AR Agr. Exp. Sta. Research Series 574:89-91. Moubarak, A. S., S. Nabhan, Z. B. Johnson, M. L. Looper, and C. F. Rosenkrans, Jr. 2009. Metabolism of ergot alkaloids by steer liver cytochrome P450 3A. AR Agr. Exp. Sta. Research Series 574:29-31. Murphy, K., M. Sales, S. Reiter, H. Brown, Jr., M. Brown, M. Looper, and C. Rosenkrans, Jr. 2009. Polymorphisms in the regulatory region of bovine cytochrome P450. AR Agr. Exp. Sta. Research Series 574:14-17. Starnes, A. R., A. H. Brown, Jr., Z. B. Johnson, J. G. Powell, J. L. Reynolds, S. T. Reiter, M. L. Looper, and C. F. Rosenkrans, Jr. 2009. Relationships between prolactin promoter polymorphisms and Angus calf temperament scores and fecal egg counts. AR Agr. Exp. Sta. Research Series 574:22-24. Chiasson, M. K., J. Carter, K. R. Bondioli, R.A. Godke and G.T. Gentry. 2009. The use of laser-assisted embryo hatching prior to the transfer of frozen-thawed in vivo-produced beef cattle embryos. LSU AgCenter Beef Cattle/Forage Res. Rep. (In press) Gentry, Jr., G.T., J.A. Roberts and R.A. Godke. 2009. Influence of age, body weight and body condition on plasma leptin concentrations in beef cattle. LSU AgCenter Beef Cattle/Forage Res. Rep. (In press) Regular speaker at the Farm & Family program for Mississippi Public Radio/National Public Radio. Topics included: Undergraduate Research in Bioinformatics, Graduate Research in Reproductive Biology