Brief Summary of Minutes of Annual Meeting

Brief Minutes of the W3171 Technical Committee Meeting

Galt Hotel, Louisville, KY

January 22, 2016

The meeting was called to order at 1 pm by Committee Chair, Jerry Bouma (Colorado). In attendance were Rick Rorie (Arkansas), Cindy Tang (Connecticut), Matt Wheeler (Illinois), Curt Youngs (Iowa), Ken Bondioli (Louisiana), Brett White (Nebraska) and Kiho Lee (Virginia).

The meeting agenda and minutes of the previous meeting were approved as read. W3171 Administrator, Milan Shipka, addressed committee members via phone since he could not attend due to recent surgery. Milan provided guidance for submission of the project's report of accomplishments, impacts and publications. He also emphasized our cooperative/collaborative work on research projects.

Station reports were presented and discussed for Arkansas, Colorado, Connecticut, Illinois, Iowa, Louisiana, Nebraska and Virginia. Potential new collaborative projects were discussed at length. To facilitate collaborative research, Jerry Bouma agreed to develop a spreadsheet on which research personnel at each station can enter their areas of interest and expertise, capabilities and techniques. This information will be used to identify overlapping research areas for additional collaborative efforts.

A motion was made and unanimously approved to recognize Bob Godke (Louisiana) for his numerous contributions to the W171/1171/2171/3171 regional research project over many years. Bob Godke passed away during 2015.

Brett White was elected to serve Secretary for the W3171 Technical Committee for 2016. Rick Rorie will serve as Chair for the coming year. The committee voted to meet in conjunction with the IETS Annual Conference in Austin, Texas in January 2017. The meeting and location will be determined at a later date.

Meeting adjourned at 4:15 pm.

Accomplishments

Objective 1:  Understand the biology and underlying mechanisms of gamete development, fertilization, and embryogenesis

Oocyte gene expression was investigated to identify markers for improved oocyte maturation in vitro.  Many deviations from normal cytoplasmic maturation were noted and maintained when maturation was delayed with Forskolin and IBMX.  Characterization of global epigenetic marks in oocytes derived from small versus large follicles suggest no difference in H3K4me3 and H3K9me3 levels, but that H3K27me3 levels may be different between oocyte from the different sized follicles.  Significant differential gene expression was noted between oocytes with different G6PDH activity that could be used as biomarkers for oocyte quality.  A system was developed for embedding and long-term culture of single porcine follicles in vitro.  Effects of follicular fluid concentrations of androstenedione on theca and granulosa cell gene expression, and the role of VEGFA isoforms in ovarian function were investigated to better understand oocyte maturation.  In vitro fertilization conditions were optimized for using sex-sorted semen for production of bovine embryos.

Investigation of sperm metabolic pathways of domestic cats demonstrated that glucose, fructose, and pyruvate are metabolized by sperm, but in a species-specific manner.   Procedures were developed for detection of GnRH-II and GnRHR-II in testicular tissue.  The role of GnRH-II in male fertility was investigated by quantifying in seminal plasma of boars, and by comparing GnRH-II receptor protein levels in the testis and epididymis, as well as the prostate, seminal vesicles and bulbourethral glands.

Viable embryos were produced in vitro, using fresh oocytes collected and transported from different countries; opening the possibility to access superior genetics and improve herds.  Also, the use of BLV seropositive donor semen for IVF showed no transmission of the virus. Progress was made in sorting embryos by an apoptotic index, then deriving meaningful gene expression and DNA methylation data.  The abundance of mRNAs for imprinted genes of embryos were determined with vast differences of these genes detected across species, suggesting these genes may play different roles in early embryo development.  Sex chromosome dosage compensation was studied in the bovine, and found that compensation occurred in both in vivo and in vitro produced embryos, as well as somatic tissues of male and female adults. 

Embryo metabolites were compared between IVF and parthenogenetically activated oocytes. Parthenogenetic embryos produced more citrate than IVF embryos, indicating citrate may be a useful marker for embryo viability.  Increased acetate and formate were detected on day 3, indicating the start of glycolysis and a possible marker for embryo viability.  Substrate use and metabolic pathway fluxes were measured in embryos, using GC-mass spectrometry.  Results demonstrate substrate balance in the media has a profound influence on pathway activities and on the routes and extent that media substrates are metabolized.  Analysis of lipids by ambient ionization mass spectroscopy allowed detection of a wide range of lipid classes in single oocytes, embryos and cell pellets, and provided information on in vitro culture impact, developmental and differentiation stages. 

Anti-Mullerian hormone and follicle counts as a predictor of superovulatory response and embryo production, confirm that both relative anti-Mullerian hormone concentration and number of small antral follicles positively correlate with embryo production.  In related studies, a successful superovulation protocol was developed for Boran cattle and the first pregnancy was established cryopreserved alpaca embryos.  The Dracula embryo manipulation devise was evaluated for use with a variety of embryos of different species, suggesting the technology is amendable to a variety of agriculturally important mammalian species.

Objective 2:  Refine methods for production of genetically modified animals to improve livestock production efficiency

The CRISPR/Cas9 system for gene editing is currently being utilized by several stations.  Targeting and genome editing constructs were designed to study LIN28B function in placenta in vivo.  In a study with the aim to use transgenic animals for bioreactors, results suggest that MAC-T cells may be a good system to evaluate gene editing via CRISPR for targeting recombinant protein expression in milk.   Preliminary data suggest that the combined cell transfection of gene target-specific CRISPR/Cas9 and RNAi to knockdown the NHEJ pathway is a viable and efficient approach to produce precise genetically modified goat donor cells carrying mono- and biallelic knock-ins of large size transgene constructs for use in cloning by SCNT.  Cloning procedures are underway using biallelic knock-in somatic cells to obtain live offspring, in order to produce and test a recombinant subunit vaccine against B. abortus.

The CRISPR/Cas9 system was used to disrupt up to three genes simultaneously at near 100% efficiency in vitro.  In vivo, efforts were successful in producing pigs lacking two genes simultaneously.  Other results demonstrate that the CRISPR/Cas9 system can be used to induce targeted mutations through NHEJ and/or HDR, and to induce homologous recombination during porcine embryogenesis.  In other efforts, sgRNAs have been designed specifically for HMGA2 and KDM1A for CRISPR Cas9 genome editing, and these are currently tested in in vitro assays. The CRISPR Cas9 genome editing tools were also used to generate mutations specifically in the trophectoderm that are being tested in vivo.

In studies focused on stem cells, derivation efficiency of induced pluripotent stem cells were improved by using the mouse model, and the roles of chromatin remodeling complexes in the reprogramming process were identified.  The effect of spatial location, current intensity, and time of exposure on the effect of a bioelectrical stimulus to change pluripotency gene expression levels in cultured fibroblast cells was analyzed.  The results could give a better understanding of optimal treatment conditions for inducing cell transformation and lead to improvement in efficiency of SCNT.

A novel Dynamic Impact Approach and functional enrichment analysis was used to show only a few pathways and functions were more induced during osteogenesis compared to adipogenesis. Some pathways were more inhibited during osteogenesis, such as cholesterol and protein synthesis. Up-stream transcription factor analysis indicated activation of several lipid-related transcription regulators (e.g., PPARs and CEBPα) during adipogenesis but osteogenesis was driven by inhibition of several up-stream regulators, such as MYC.  Between MSCs the data indicated an ‘adipocyte memory’ in ASC with also an apparent lower immunogenicity compared to BMSC during differentiation. Overall the analysis allowed proposing a dynamic model for the adipogenic and osteogenic differentiation in porcine ASC and BMSC. 

Porcine chondrocytes and ASC were able to form cartilage matrix in a pellet culture system with chondrogenic media regardless of additive, while porcine fibroblasts and periosteal cells showed limited to no chondrogenic potential in the conditions tested. Chondrocyte pellet growth is due to extracellular matrix deposition.  Another study demonstrated a modular platform approach founded on 3D printing, for developing tissue engineering therapies and illustrated the design control process for modular implementation of two scaffold based tissue engineering therapies: airway reconstruction and bone tissue engineering based spine fusion.

Tagged versions (His, Myc or Flag tags) were generated for the most promising antigens of different zoonotic organisms including; Tuberculosis, Brucellosis, Leptospirosis, Q fever, Trypanosomiasis, Rabies and Porcine Cysticercosis. Sequences encoding these antigens were cloned into a replication defective adenoviral expression vector.  Recombinant proteins were produced in two different cell lines (HEK 293 and MAC-T), cell lysates and supernatants were examined for tagged proteins using western blot analysis. Constructs are being validated for production of recombinant proteins in the mammary gland for vaccine development.

In efforts directed toward cryopreservation, it was shown that cleavage rates post-thaw and fertilization were similar when oocytes were vitrified before or after vitrification.  The Mouse Dracula tool was used to demonstrate that mouse embryos at the early-, expanded-, or even the hatched-stage can effectively have their coleomic contents exchanged with cryoprotectant. These results could improve cryopreservation efforts and make these procedures less dependent on temperature-change schedules.  Efforts directed toward storage of ram semen demonstrated that regardless of semen extender or storage temperature, fresh ram semen was found to loose progressive motility within 24 hours of storage.

W-3171 Publications

2014

Chen H, Fu J, Chen H, Hu Y, Soroka DN, Prigge JR, Schmidt EE, Yan F, Major MB, Chen X, Sang S. 2014.  Ginger compound [6]-shogaol and its cysteine-conjugated metabolite (M2) activate Nrf2 in colon epithelial cells in vitro and in vivo. Chem Res Toxicol. 2014 Sep 15;27(9):1575-85.

Godke, RA, M Sansinena, and CR Youngs. 2014.  Assisted reproductive technologies and embryo culture methods for farm animals. In (CA Pinkert, Ed.) Transgenic Animal Technology: A Laboratory Handbook, 3^rd Ed., Elsevier, London, pp. 581-638.  DOI: http://dx.doi.org/10.1016/B978-0-12-410490-7.00022-0

Vivanco, W, D Ponce, M Miguel, C Youngs, E Huamán, S. León, and M Asparrin.  2014.  Repeatability of the ovarian response and embryo production in superovulated Huacaya alpacas. Spermova 4(1):77-79.

Youngs CR. 2014. Embryo development and survival in ruminant livestock species. In (G. Gutierrez and M Rothschild, Eds.). Proceedings of the International Seminar on Advances in Animal Production in Latin America, November 5-7, 2014, Lima Peru, pp. 40-51.

2015

Abedal-Majed, M.A., M.P. Magamage, R.J. Vinton, R.A. Cushmann, A.K. McNeel, H. Freetly, J.R. Wood, and A.S. Cupp. 2015. Vascular Endothelial Growth Factor A 165 (VEGFA165), angiogenic isoform, promotes while VEGFA165b antagonizes VEGFA165 stimulated follicular progression in bovine ovarian cortical pieces cultured from pre-pubertal heifers. Society for the Study of Reproduction 47^th Annual Meeting. San Juan, Puerto Rico. p. 246.

Bionaz, M., Monaco, E.  and Wheeler, M.B. 2015. Transcription Adaptation during In Vitro Adipogenesis and Osteogenesis of Porcine Mesenchymal Stem Cells: Dynamics of Pathways, Biological Processes, Up-Stream Regulators, and Gene Networks. PLoS One. 2015 Sep 23;10(9):e0137644. doi: 10.1371/journal.pone.0137644. eCollection 2015.

Bondioli, K.R.  Cryopreservation of Bovine Embryos. 2015.  In: Hopper, R. (ed) Bovine Reproduction. John Wiley and Sons, pp. 718-722.

Campos-Chillon F, Farmerie TA, Bouma GJ, Clay CM, Carnevale EM. 2015. The effect of aging on gene expression and mitochondrial DNA in the equine oocyte and follicle. Reprod Fertil & Develop, Mar 19. doi: 10.1071/RD14472. [Epub ahead of print].

Canovas S and Ross PJ. 2015.  Mechanisms of epigenetic remodeling during preimplantation development. Reproduction, Fertility and Development. 28(2): 25-40.

Cebula M, Schmidt EE, Arnér ES.  2015.  TrxR1 as a potent regulator of the Nrf2-Keap1 response system.. Antioxid Redox Signal. 23(10):823-53.

Cederberg, R.A., J.E. Smith, E.A. McDonald, C. Lee, A.R. Perkins, and B.R. White. 2015. Activity of the porcine gonadotropin-releasing hormone receptor gene promoter is partially conferred by a distal gonadotrope specific element (GSE) within an upstream enhancing region, two proximal GSEs and a retinoid X receptor binding site. Reprod. Biol. Endocrinol. 13:45.

Center, K., D. Dixon, and R. Rorie.  2015.  Anti-mullerian hormone and follicle counts as predictors of superovulatory response and embryo production in beef cattle. Arkansas Agri. Exp. Sta. Res. Series 628:31-33.

Cleys ER, Halleran JL, Enriquez VA, da Silveira JC, West RC, Winger QA, Anthony RV, Bruemmer JE, Clay CM, Bouma GJ. 2015. Androgen Receptor and Histone Lysine Demethylases in Ovine Placenta. PLoS ONE 10(2):e0117472. [PMID: 25675430]

Cox L, Hintze KJ, and ISOM SC.  June 2015.  Impact of a Total Western Diet on oocyte quality in swine.  48th Annual Meeting of the Society for the Study of Reproduction.  San Juan, PR. 

Cox L, ISOM SC, and Hintze KJ.  April 2015.  Formulation of the Total Western Diet (TWD) for Pigs and Phenotypic Differences After a 12-week Feeding Trial.  29th Annual Meeting of the Federation of American Societies for Experimental Biology.  Boston, MA. 

Cushman, R.A., R.G. Tait Jr., A.K. McNeel, E.D. Forbes, O.L. Amundson, C.A. Lents, A.K. Lindholm-Perry, G.A. Perry, J.R. Wood, A.S. Cupp, T.P. Smith, H.C. Freetly, and G.L. Bennett. 2015. A polymorphism in myostatin influences puberty but not fertility in beef heifers, whereas µ-calpain affects first calf birth weight. J. Anim. Sci. 93:117-26.

da Silveira JC, Winger QA, Bouma GJ, Carnevale EM. 2015.  Age effects on follicular fluid exosomal miRNAs and TGFβ signaling during follicle development in the mare. Reprod Fertil & Develop, May 7. doi: 10.1071/RD14452. [Epub ahead of print].

Davis, A.J., T.D. Lester, E.A. Backes and R.W. Rorie.  2015.  Sequential use of Estrotect estrous detection patches as a reproductive management tool.  Prof. Anim. Sci. 31:50-56. 

Davis, A.J., T.D. Lester, E.A. Backes and R.W. Rorie.  2015.  Serial use of Estrotect estrous detection patches as a reproductive management tool.  J. Anim. Sci., So. Sec. (abstr 38, pg 16)

Desaulniers, A.T., R.A. Cederberg, G.A. Mills, C.A. Lents and B.R. White. 2015. A putative role for GnRH-II and its receptor in spermatogenic function of boars. Society for the Study of Reproduction 47^th Annual Meeting. San Juan, Puerto Rico. p. 210.

Desaulniers, A.T., R.A. Cederberg, G.A. Mills, J.J. Ford, C.A. Lents, and B.R. White. 2015. LH-independent testosterone secretion is mediated by the interaction between GNRH2 and its receptor within porcine testes. Biol. Reprod. 93:45.

Diaz, FA, EJ Gutierrez, BA Foster, PT Hardin and KR Bondioli.  2015.  Effect of the Repeated Use of Open System Vitrification Devices on MII Stage and Cleavage Rates of Bovine Cumulus-oocyte Complexes.  Reproduction, Fertility and Development 28:145.

Dodson, M.V.,  R.E. Allen, M. Du, W.G. Bergen, S.G. Velleman, S.P. Poulos, M. Fernyhough-Culver, M.B. Wheeler, S.K. Duckett, M.R.I. Young, B.H. Voy, Z. Jiang and G.J. Hausman, 2015. Invited Review: Evolution of meat animal growth research during the past 30 years: Adipose and muscle stem cells. J. Anim. Sci. 2015.93:457–481.

Douthit, C.J., R.A. Cederberg, A.T. Desaulniers, J.M. Kouba, T.L. Douthit, D.M. Grieger and B.R. White. 2015. Identification of GnRH isoforms and their cognate receptors in the equine testis. Equine Science Society Annual Meeting. St. Pete Beach, Florida. p. 138.

Ebramzadeh, E., Sangiorgio, S.N., Wheeler, M.B.,  Green, G.E. 2015. Design Control for Clinical Translation of 3D Printed Modular Scaffolds. Annals of Biomedical Engineering, Vol. 43, No. 3,774–786.

Enriquez VA, Cleys ER, da Silveira JC, Spillman MA, Winger QA, Bouma GJ. 2015. High LIN28A Expressing Ovarian Cancer Cells Secrete Exosomes That Induce Invasion and Migration in HEK293 Cells. BioMed Res Int 2015, Article ID 701390.

Eriksson S, Prigge JR, Talago EA, Arnér ES, Schmidt EE. 2015.  Dietary methionine can sustain cytosolic redox homeostasis in the mouse liver.  Nat Commun. 6:6479.

Eriksson, S, and Schmidt, E.E.  2015.  “Thioredoxin reductase: A coordinator of metabolic activities”. In “Selenium, its molecular biology and role in human health”. Edited by Brigelieus-Flohe, R. Within series entitled: “Oxidative stress in health and disease”. CRC Press.

Ferré LB, Bogliotti Y, Chitwood JL, Rodríguez CF ,Ortega HH, Kjelland ME and Ross PJ.  2015. Comparison of different fertilization media for an in vitro maturation/fertilization/culture system using flowcytometrically sorted X-chromosome-bearing sperm for bovine embryo production. Reproduction, Fertility and Development. May 13 Epub ahead of print.

Ferreira^, C.R., Jarmusch, A.K., Pirro, V., Alfaro, C.M., González-Serrano, A.F., Niemann, H., Wheeler, M.B., Rabel, R.A.C., Hallett, J.E., Kaufmann, A. and Cooks, R.G.  2015.  Ambient Ionization Mass Spectrometry for Lipid Profiling and Structural Analysis of Oocytes, Preimplantation Embryos and Stem Cells. Reproduction, Fertility and Development, 27:621–637.

Foster, BA, FA Diaz, PT Hardin, EJ Gutierrez and KR Bondioli.  2015.  Characterization of Bovine Oocyte Cytoplasmic Maturation with Common IVM Protocols.  Reproduction, Fertility and Development  28:232

Hatzel JN, Bouma GJ, Cleys ER, Bemis LT, Ehrhart EJ, McCue PM. 2015. Identification of heat shock protein 10 within the equine embryo, endometrium, and maternal peripheral blood mononuclear cells. Theriogenology 83(5):832-839. [PMID: 25542459]

Herzog, K.K., Milner, D.J., Johnson, S.J., Wheeler, M.B.  2015. Comparison of Chondrogenic Potential of Porcine Adipose-Derived Stem Cells, Chondrocytes, Periosteal Cells, and Fibroblasts in a Pellet Culture System. Reproduction, Fertility and Development 27(1) 253-253.

Hollister, S.J., Flanagan, C.L., Zopf, D.A., Morrison, R.J., Nasser, H.,  Patel, J.J., Hughes JR, Berger T.  2015.  Testicular fluid regulates apical blebbing in the porcine epididymis. American Society of Andrology.

Hughes JR, Berger T. 2015. Development of apical blebbing in the boar epididymis. PLOS One 2015.

Jahnke MM, JK West, and CR Youngs.  2015.  Evaluation of in vivo derived bovine embryos. In (RM Hopper, Ed) Bovine Reproduction, Wiley & Sons, Hoboken NJ, pp 733-748.

Jiang Z, Harrington P, Zhang M, Marjani SL, Park J, Kuo L, Pribenszky C, Tian XC. 2015. Effects of High Hydrostatic Pressure on Expression Profiles of In Vitro Produced Vitrified Bovine Blastocysts (accepted by Scientific Reports, Dec 2015).

Jiang Z, Tang Y, Zhao X, Zhang M, Donovan DM, Tian XC. 2015.  Knockdown of Brm and Baf170, Components of Chromatin Remodeling Complex, Facilitates Reprogramming of Somatic Cells. Stem Cells and Development 24:2328-36.

Katleba K, Legacki EL, Conley AJ, Berger T.  2015.  Steroid Regulation of Early Postnatal Development in the Corpus Epididymidis of Pigs. Journal of Endocrinology 225(3): 125-134.

Klohonatz KM, Hess AM, Hansen TR, Squires EL, Bouma GJ, Bruemmer JE. 2015. Equine endometrial gene expression changes during and after maternal recognition of pregnancy. J Anim Sci 93(7):3364-3376.

Lambe-Steinmiller, J.C.  2015.  The Use of Assisted Reproductive Technologies to Improve Pregnancy Rates in White-tailed Deer.  Louisiana State University. PhD dissertation

Legacki E, Conley AJ, Nitta-Oda BJ, Berger T.  Porcine Sertoli Cell Proliferation after Androgen Receptor Inactivation. Biology of Reproduction. 2015; 92(4): 93.

Li Q, Suasnavas E, Heywood S, Xiao L, Zhou A, Isom SC.  2015.  Biochemical, biophysical and genetic changes of porcine trophoblast-derived stem-like cells during differentiation as evaluated using Raman microspectroscopy, Atomic force  microscopy and quantitative polymerase chain reaction. Genesis. 2015 Oct 28. doi: 10.1002/dvg.22907. [Epub ahead of print] PubMed PMID: 26509257

Li Q, Suasnavas E, Williams S, Li R, ISOM SC, and Zhou A.  2015  Atomic force microscopy (AFM), Raman Microspectroscopy (RM) and gene chip monitoring of porcine trophoblast derived cell differentiation.  SPIE BiOS Conference on Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications XII.  San Francisco, CA

Meyer, L., T. Devine, M. Looper, D. Philipp, D. Hubbell, III, R. Rorie, and C. Rosenkrans, Jr.  2015.  Associations between heifer endocrine profiles and reproductive efficiency.  Arkansas Agri. Exp. Sta. Res. Series 628:38-39.

Nemere I, Garbi N, Winger Q. 2015. The 1,25D3 -MARRS receptor/PDIA3/ERp57 and lifespan. J Cell Biochem. 116(3):380-5. doi: 10.1002/jcb.24986. PMID: 25283641

Non-Invasive Quantification of Cartilage Using a Novel In Vivo Bioluminescent Reporter Mouse. Mailhiot SE, Zignego DL, Prigge JR, Wardwell ER, Schmidt EE, June RK. 2015.  PLoS One. 2015 Jul 7;10(7):e0130564.

Park JP, Lai L, Samuel MS, Wax D, Prather RS, Tian XC. 2015. Disruption of mitochondria-to-nucleus interaction in deceased cloned piglets. PLoS ONE 2015

Prigge JR, Hoyt TR, Dobrinen E, Capecchi MR, Schmidt EE, Meissner N. 2015. Type I IFNs Act upon Hematopoietic Progenitors To Protect and Maintain Hematopoiesis during Pneumocystis Lung Infection in Mice.  J. Immunol. 195(11):5347-57.

Romereim, S., A.F. Summers, B.E. Pohlmeier, R.M. McFee, R. Spuri-Gomes, S.G. Kurz, A.K. McNeel, R.A. Cushmann, J.S. Davis, J.R. Wood, and A.S. Cupp. 2015. Granulosa cell cycle regulation and steroidogenesis in a high androstenedione follicular microenvironment. Society for the Study of Reproduction 47^th Annual Meeting. San Juan, Puerto Rico. p. 245.

Rubessa, M., Ambrosi, A., Polkoff, K.M., Stewart, J.W., Herzog^, K.K., Denmark, S.E. Wheeler, M.B. 2015. Metabolic Profiling of Male and Female Bovine Embryos Using Nuclear Magnetic Resonance (NMR) Imaging. Reproduction, Fertility and Development 27(1) 158–159.

Rubessa, M., Herzog^, K.K., Ambrosi, A., Stewart, J.W., Polkoff, K.M., Denmark, S.E. Wheeler, M.B. 2015. Non-Invasive Analysis of Embryo Metabolites Using Nuclear Magnetic Resonance. Reproduction, Fertility and Development  27(1) 157–157.

Rubessa, M., Nasser, L.F., Wheeler, M.B. 2015. Practical Applications of In Vitro Embryo Production in Latin America. Proceedings 11^th Simposium Internacional de Reproduction Animal, Cordoba, AR.

Rutigliano HM, Wilhelm A, Hall J, Shi B, Meng Q, Stott R, Bunch TD, White KL, Davies CJ, Polejaeva IA. Cytokine gene expression at the maternal-fetal interface after somatic cell nuclear transfer pregnancies in small ruminants. Reprod Fertil Dev. 2015 Oct 14. doi: 10.1071/RD15103. [Epub ahead of print] PubMed PMID: 26463834.

Ryu J and Lee K.  2015. Complete disruption of RAG2 gene during porcine embryogenesis utilizing CRISPR/Cas9 system. Biol Reprod. (Suppl. 1): 377.

Sanz-Fernandez MV, Johnson JS, Abuajamieh M, Stoakes SK, Seibert JT, Cox L, Kahl S, Elsasser TH, Ross JW, ISOM SC, Rhoads RP, Baumgard LH.  2015.  Effects of heat stress on carbohydrate and lipid metabolism in growing pigs.  Physiol Rep. 3(2) . pii: e12315. doi: 10.14814/phy2.12315. Print 2015 Feb 1.  PMID: 25716927

Sargent, K.A., R. Spuri-Gomes, J.R. Essink, S.G. Kurz, W.E. Pohlmeier, N. Ferrara, G.J. Bouma, D.J. McLean, and A.S. Cupp.  2015. Sertoli cell-specific elimination of Vegfa impairs spermatogonial stem cell (SSC) maintenance and reduces fertility in male mice. Society for the Study of Reproduction 47^th Annual Meeting. San Juan, Puerto Rico. p. 266-267.

Sargent, K.M., N. Lu, D.T. Clopton, W.E. Pohlmeier, V.M. Brauer, N. Ferrara, D.W. Silversides, and A.S. Cupp. 2015. Loss of vascular endothelial growth factor A (VEGFA) isoforms in granulosa cells using pDmrt-1-Cre or Amhr2-Cre reduces fertility by arresting follicular development and by reducing litter size in female mice. PLoS One. 10:e0116332.

Sargent, K.M., R.M. McFee, R. Spuri Gomes, and A.S. Cupp. 2015. Vascular endothelial growth factor A: just one of multiple mechanisms for sex-specific vascular development within the testis? J. Endocrinol. 227:R31-50.

Schmidt EE.  2105.  Interplay between cytosolic disulfide reductase systems and the Nrf2/Keap1 pathway. Biochem Soc Trans. 43(4):632-8.

Spuri-Gomes, R., S. Tenley, M.A. Abedal-Majed, S.G. Kurz, J.R. Wood, J. Bergman, and A.S. Cupp. 2015. Cows with intrafollicular androgen excess have lower sex hormone binding globulin and appear to be chronic or sporadic anovulatory. Society for the Study of Reproduction 47^th Annual Meeting. San Juan, Puerto Rico. p. 250.

Suasnavas E, Heywood S, Ward A, Cox L, O’Grady M, Zhao Y, and ISOM SC.  Isolation and characterization of trophoblast-derived stem-like cells from peri-implantation porcine embryos. Anim Reprod Sci. 2015 Mar; 154:128-41. doi: 10.1016/j.anireprosci.2015.01.012. Epub 2015 Jan 31.  PMID: 25660622. 

Szerlong HJ, Herman JA, Krause CM, DeLuca JG, Skoultchi A, Winger QA, Prenni JE, Hansen JC. 2015. Proteomic characterization of the nucleolar linker histone H1 interaction network. J Mol Biol. 427(11):2056-71. doi:10.1016/j.jmb.2015.01.001. Epub 2015 Jan 10. PMID: 25584861

Talbot, H.A., P. Zhang, X. Hou, E. Keiser, F. Yu, A.S. Cupp, and J.S. Davis. 2015. Identification of IL-17 as a potential mediator of luteolysis: Inhibition of progesterone secretion. Society for the Study of Reproduction 47^th Annual Meeting. San Juan, Puerto Rico. p. 243.

Tanphaichitr N,  Kongmanas K,Kruevaisayawan H, Saewu A, Sugeng C, Fernandes J, Souda P, Angel JB, Faull KF, Aitken RJ, Whitelegge J, Hardy D, Berger T, Baker M. 2015. Remodeling of the plasma membrane in preparation for sperm-egg recognition: roles of acrosomal proteins. Asian Journal of Andrology 17(4): 1-9.

Thorson, J.F., A.T. Desaulniers, C. Lee, B.R. White, J.J. Ford, C.A. Lents. 2015. The role of RFamide-related peptide 3 (RFRP3) in regulation of the neuroendocrine reproductive and growth axes of the boar. Anim. Reprod. Sci. 159:60-65.

Tian XC. 2015. Establishing the standards of gene expression during bovine pre-implantation development. (presented in the 12^th Annual meeting of the Asian Reproductive Biotechnology Society; November 26-29, 2015, Hanoi, Vietnam). P.8

Vivanco-Mackie HW, MDP Salazar, M Miguel, C Youngs, and M Asparrin.  2015.  Embryo survival to calving according to type of embryo and embryo transfer method in alpacas. Reprod Fertil Dev 27(1):173 (abstr).

Wheeler, M.B., Kim, D., Beebe, D. US Patent "Microfluidic Systems and Methods", patent # 9,157,550, issued October 13, 2015

White, B.R. 2015.  Can a “reproductive hormone” predict appetite in first parity, lactating sows? Nebraska Pork Talk, Nebraska Pork Producers Association. 47(3):6-7.

Xie, F., C.L. Anderson, K.R. Timme, S.C. Fernando, and J.R. Wood. 2015. Increased firmicutes in the cecum of obese female mice is correlated with increased Pou5f1 and Dppa3 mRNAs in growing oocytes. Society for the Study of Reproduction 47^th Annual Meeting. San Juan, Puerto Rico. p. 248.

Youngs CR and KL Klemesrud. 2015. Update on bovine pregnancy testing technologies [Actualización sobre tecnologías de pruebas de gestación bovina]. Proceedings of the 38^th Annual Scientific Meeting of the Peruvian Association of Animal Production, October 7-9, Ayacucho, Peru, pp. 23-28.

Youngs CR, HW Vivanco-Mackie, DP Salazar, M Miguel, M Asparrin. 2015. Recent research on the cryopreservation of preimplantation embryos from South American camelids [Investigaciones recientes sobre la criopreservación de embriones de preimplantación de camélidos sudamericanos]. Proceedings of the 38th Annual Scientific Meeting of the Peruvian Association of Animal Production, October 7-9, Ayacucho, Peru, pp. 18-22.

Youngs CR. 2015. Importance of animal production in global food security [Importancia de la producción animal en la seguridad alementaria mundial]. Proceedings of the 38th Annual Scientific Meeting of the Peruvian Association of Animal Production, October 7-9, Ayacucho, Peru, pp. 14-17.

2016

Acharya, A., J.M. Burke, K. Coffey, and R.W. Rorie.  2016.  Effect of semen extender and storage temperature on ram sperm motility over time.  J. Anim. Sci., So. Sec. (accepted).

Center, K., D. Dixon, and R. Rorie.  2016.  Anti-mullerian hormone as a predictive endocrine marker for superovulatory response and embryo production in beef cattle.  Reprod. Fertil. Dev. (in press)

Cox L, Vanderwall DK, Parkinson KC, Sweat L, and ISOM SC.  Expression profiles of select genes in cumulus-oocyte complexes from young and aged mares.  Reprod Fertil Dev. IN PRESS.  2015 May 15.  doi:  10.1071/RD14446.  PMID:  25976356

Duan JE, Jue NK, Jiang Z, O’Neill R, Wolf E, Blomberg LA, Dong H, Zheng X, Chen J, and Tian XC. 2015. (to be presented in the 42nd Annual Meeting of the International Embryo Transfer Society, Louiville, Kentucky, USA Jan 22-26, 2016).

Jiang Z, Dong H, Zheng X, Donovan DM, Chen J, Tian XC. 2015. mRNA Levels of Imprinted Genes in Bovine In Vivo Oocytes, Embryos and Cross Species Comparisons in Humans, Mice and Pigs (accepted by Scientific Reports, Nov 30, 2015)

Jiang Z, Harrington P, Zhang M, Marjani SL, Park J, Kuo L, Pribenszky C, Tian XC. 2015. Effects of high hydrostatic pressure on expression profiles of in vitro produced, vitrified bovine blastocysts. (to be presented in the 42nd Annual Meeting of the International Embryo Transfer Society, Louiville, Kentucky, USA Jan 22-26, 2016).

Johnson, S., Milner, D., Lopez-Lake, H., Wheeler, M.B.  2016. Chondrogenic pellet cultures for cartilage tissue engineering grow by deposition of matrix and not by cellular proliferation. Reproduction, Fertility and Development, 28, 254-254.

Long JA, HD Blackburn, A Martin, FG Silversides, R Taylor, Jr, and CR Youngs.  2016. The need for agricultural innovation to sustainably feed the world by 2050: Protecting food animal gene pools for future generations. CAST Publication (under review).

Lotti, S.N., Tasan, I., Zhao, H., Wheeler, M.B. 2016.  MAC-T cells as a tool to examine genome editing using clustered regularly interspaced short palindromic repeats (CRISPR). Reproduction, Fertility and Development, 28, 251-251.

Polkoff, K.M., Rubessa, M., Winters, R., Lopez, N.A., Wheeler, M.B. 2016. Comparison of two embedding systems for follicular culture. Reproduction, Fertility and Development, 28, 192-192.

Rodriguez, S.J.R. Ramirez, Y.E., Gomes Jr., E., Nasser, L.F., Pontes, J.H.F., Rubessa, M., Wheeler, M.B. 2016. Comparison of in vitro embryo production in Panama using Holstein oocytes with Bos indicus sexed semen from donors in different locations: field data. Reproduction, Fertility and Development, 28,183-183.

Rorie, R.W., C.L. Williams, and T.D. Lester.  2016.  Association of osteopontin gene promoter single nucleotide polymorphisms with bull semen quality.  Adv. Reprod. Sci. (accepted).

Rubessa, M., Ambrosi, A., Polkoff, K.M., Denmark, S.E., Wheeler, M.B. 2016. Embryo sugar consumption during in vitro development. Reproduction, Fertility and Development, 28, 175-175.

Rubessa, M., Rocha, R., Lima, L., Winters, R., Figueiredo, J.R., Wheeler, M.B. 2016. Comparison of NCSU-23 and alpha-minimal essential media in the development of isolated porcine preantral follicles in vitro. 2016. Reproduction, Fertility and Development, 28,198-198.

Sommer MM and CR Youngs. 2016. Impact of embryo transfer technology on the production of artificial insemination sires for the US dairy cattle industry. Animal Industry Report, Iowa State University (submitted).

Stewart, J., Rubessa, M., Polkoff, K.M., Lotti, S.N., Wheeler, M.B. 2016. Risk of transmission of bovine leukosis virus (BLV) using seropositive bulls for in vitro fertilization embryo production. Reproduction, Fertility and Development, 28, 255-255.

Tavares, K.C.S., Lazzarotto, C.R., Neto, S.G., Martins, L.T., Aguiar, L.H., Calderon, C.E.M., Teixeira, L.P.R., Lopes, F.E.M., Wheeler, M.B., Long, C.R., Whitelaw, B., Bertolini, M., Bertolini, L.R. 2016. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 associated with transient depletion of non-homologous end-joining pathway increased gene-targeting efficiency in goat fibroblasts. Reproduction, Fertility and Development, 28, 252-252.