Brief Summary of Minutes of Annual Meeting

Accomplishments

Objective 1: Identify Mechanisms that Regulate Ovarian Function and Oocyte Quality during the Estrous Cycle

Demonstrated that orphan nuclear receptor steroidogenic factor 1 (SF-1; Nr5a1) is crucial to the regulation of the primordial follicle pool, regulating assembly and survival of primordial follicles, and their eventual exit to join the growing pool of follicles.

Generated results that may indicate that in mitochondria as at the plasma membrane, PA-6 potassium channels might be involved in hyperpolarization. Novel insights into potassium homeostasis in maturing mouse oocytes. Deeper exploration of the roles played by potassium and potassium channels in oocyte maturation could pave the way for their potential application in treating infertility and/or developing better reproductive strategies in animals.

Demonstrated that Zrt- and Irt-like Protein 9 (ZIP9) is not hormonally regulated in the mouse ovary and the ZIP9 agonist, epicatechin, does not increase intracellular zinc. These results demonstrate that ZIP9 protein and signaling may be regulated differently in mammalian ovaries compared to other cell types in which ZIP9 has been characterized.

Demonstrated that use of multiple FSH administrations enhances ovarian response and oocyte developmental competence leading to greater embryo yield in Holstein heifers.

Demonstrated that the occurrence of endometritis after induced pregnancy loss did not affect the lifespan of the CL after pregnancy loss during the second month of pregnancy in cattle.

Determined the effect of Ang II on CCN1 expression, and the activity of gelatinase MMPs (MMP2 and MMP9), in steroidogenic cells obtained from the bovine corpus hemorrhagicum CL.

Determined the effect of calcium ionophore on CCN1 expression, and the activity of gelatinase MMPs (MMP2 and MMP9) by steroidogenic cells obtained from the bovine corpus hemorrhagicum CL.

Determined the effect of phytoestrogens in red clover on progesterone biosynthesis in bovine CL. Phytoestrogens present in red clover may have a negative effect on the ability of LH to stimulate progesterone production in luteal tissue.

Generated data that suggest that Ang II and calcium, both of which play roles in coagulation and healing of wounds, such as the one found on the ovarian surface after ovulation, may also increase the expression of the angiogenic inducer, CCN1, which is highly expressed in the early, developing bovine corpus hemorrhagicum CL.

Determined that KGN cells, an immortal granulosa cell tumor line, express proteins that are post translationally modified by O-GlcNAcylation and that manipulation of O-GlcNAcylation in vitro impairs KGN proliferation and oxidative phosphorylation.

Demonstrated that manipulation of O-GlcNAcylation in primary cultures of bovine granulosa cells impairs transcript expression of STAR, but otherwise has no effect on expression of FSHR and CYP19A1 transcripts, or estradiol production.

Provided preliminary evidence that induction of hypoxia in primary cultures of bovine granulosa cells augments estradiol production; however, manipulation of O-GlcNAcylation does not impact this.

Completed preliminary experiments evaluating the effects of hyper-O-GlcNAcylation and hypo-O-GlcNAcylation on the proteome/glycome of bovine granulosa cells.

Explored how the interleukin-6 (IL6) cytokine family regulates oocyte maturation, early embryo development, blastocyst formation, and early conceptus development in cattle. Demonstarted that supplementing IL6 improves bovine conceptus elongation.

Investigated how antioxidant supplementation may improve oocyte and embryo development potential in cattle. 

Determined the effect of form of selenium on serum blood metabolites, observing changes that suggest treatment-induced effects on hepatic function.

Demonstrated that the ovarian function and endocrine phenotypes of lactating dairy cows during the estrous cycle are associated with genomic-enhanced predictions of fertility potential. Cows in the lowest tertile for genomic daughter pregnancy rate were more likely to have aberrant estrous cycle phenotypes, had smaller corpus luteum size and produced less progesterone during the luteal phase.

Objective 2: Determine Factors Associated with Fertilization, Embryo Development, and Conceptus-Endometrial Interactions that Dictate Pregnancy Success

Identified the orphan nuclear receptor SF-1 as a critical regulator of the formation of the ovarian reserve of follicles.

Identified a potasium channel family (Kir2.X) important for COC maturation.

Determined the that PMSG and hCG had no effect in the regulaiton of ZIP9 protein and that hte ZIP9 agonist, epicatechin had no effect on zinc transport in mammalian granulosa cells.

Determined that exposure of bovine oocytes to H2O2 during IVM culture caused reduced blastocyst development and altered abundance of METTL3 and FTO. Taken together, these data contribute to the knowledge regarding m6A-mediated mRNA metabolism to enhance our understanding of its role in the oocyte-to-embryo transition (OET) across species.

Demonstrated that overall decreases in mRNA abundance in placenta from obese dams could cause global effects of maternal obesity on synthesis, stability, and/or mRNA translation and represents a novel target for reversing abnormal placentation associated with obesity.

Objective 3: Develop and Evaluate Novel Reproductive Management Strategies and Technologies to Improve Reproductive Performance of Ruminants

Demonstrated that presynchronization strategies that do not require progesterone (P4) withdrawal, such as the 6&6 or 8&6 Synch, provide a more user-friendly alternative with comparable fertility to traditional protocols.

Generated data that suggested a modest increase in serum vitamin A and D will improve pregnancy rates after AI.

Demonstrated that perfusion techniques to vitrify whole ovaries are highly successful in preserving follicular function in ewes.

Evaluated the accuracy of a lateral flow-based electronic system for determination of the pregnancy and ovarian status of cattle.

Determined the effect of a targeted reproductive management programs based on automated detection of estrus during the voluntary waiting period on reproductive performance of lactating dairy cows

Provided evidence that combining reproductive outcomes predictors and automated estrus alerts recorded during the voluntary waiting period identified subgroups of cows with different reproductive performance potential.

Demonstrated that delayed induction of ovulation affects the ovarian function of lactating dairy cows synchronized with the Double-Ovsynch protocol and increases insemination after detected estrus.

Developed novel reproductive management programs aimed at increasing fertility of first service through promotion of expression of estrus at the end of synchronization of ovulation protocols.

Peer-reviewed journal articles reporting research from this project in 2023-2024

Hughes CHK, Smith OE, Meinsohn MC, Brunelle M, Gévry N, Murphy BD. Steroidogenic factor 1 (SF-1; Nr5a1) regulates the formation of the ovarian reserve. Proceedings of the National Academy of Sciences 2023; 120:e2220849120. https://doi.org/10.1073/pnas.2220849120

Pate JL, Hughes CHK. Luteal prostaglandins: mechanisms regulating luteal survival and demise in ruminants. Animal 2023; 17:100739. 10.1016/j.animal.2023.100739

Hughes CHK and Murphy BD. Steroidogenic factor 1 (SF-1; Nr5a1) orchestrates the establishment and depletion of the ovarian reserve. Society for the Study of Reproduction, 56th annual meeting. July 11-14, 2023, Ottawa, Ontario, Canada. (Invited)

Krause, ART, Garner, TB and Diaz FJ. Inhibition of mouse oocyte maturation by a specific IK1 inhibitor. Society for the Study of Reproduction, July 11-14, 2023, Ottawa, Canada.

Carothers A and Diaz FJ. Hormonal Regulation and Zinc Influx Activity of Zrt- and Irt-like Protein 9 (ZIP9) in the Mammalian Ovary. Society for the Study of Reproduction, July 11-14, 2023, Ottawa, Canada.

Snider AP, Spuri-Gomes R, Summers AF, Tenley SC, Abedal-Majed MA, McFee RM, Wood JR, Davis JS, and Cupp AS (2023) Identification of lipids and cytokines in plasma and follicular fluid before and after FSH stimulation as potential markers for follicular maturation in cattle. Animals, 13:3289, PMID: 37894013

Ermisch AF, Bidne KL, Kurz SG, Bochantin KA, and Wood JR (2023) Ovarian inflammation mediated by TLR-4 increased transcripts of maternal effect genes and decreased embryo development. Biol Reprod 108(3): 423-436, PMID: 36461933

Ermisch AF and Wood JR (2023) N6-methyladenosine dynamics and differential methylation of maternal and zygotic mRNAs during the early stages of the maternal to zygotic transition. 56th Annual Meeting of the Society for the Study of Reproduction, Ottawa, Canada, Pre-doctoral Platform Competition Talk

Monteiro P. L. J., Consentini C. E. C., Andrade J. P. N., Beard A. D., García-Guerra A., Sartori R., Wiltbank M. C. 2023. Research on timed AI in Beef Cattle: Past, Present and Future, a 27-year Perspective. Theriogenology, 211: 161-171. DOI: 10.1016/j.theriogenology.2023.07.037.

Hayden C. B., Sala R. V., Pereira D. C., Moreno J. F., García-Guerra A. Use and dosage of p-FSH for ovarian superstimulation before ovum-pick up and in vitro embryo production in pregnant Holstein heifers. Journal of Diary Science, 106: 8110-8121. DOI: 10.3168/jds.2023-23576.

Carranza Martin A, García-Guerra A, Relling A. 2023. Effects of polyunsaturated fatty acid supplementation on plasma and follicular fluid Resolvin D1 concentration and the expression of mRNA of genes associated with resolving synthesis in ewes. Journal of Animal Science, 101: skad310.  DOI: 10.1093/jas/skad310.

Constantino, J. V., Carranza-Martin A., Premanandan C., Kirkpatrick B. W., Wiltbank M. C., García-Guerra A. 2023. Preantral follicle numbers and size in heifers carrying Trio, a bovine high fecundity allele. Reproduction, 166: 13-26. DOI: 10.1530/REP-23-0072.

Rojas Canadas E., Battista S.E., Kieffer J.D., Wellert S.R., Mussard M. L., Garcia-Guerra A. 2023. GnRH dose at initiation of a 5-day CO-Synch + P4 for fixed time artificial insemination in suckled beef cows. Animal Reproduction Science, 250: 107210.  DOI: 10.1016/j.anireprosci.2023.107210.

2023. Rykaczewski, N. B. Bello, J. D. Kieffer, A. Z. Chang, M. Trotter, A. Garcia-Guerra. 2023. “Evaluation of on-animal sensors for monitoring diurnal physical activity of beef bulls during the breeding season” Journal of Animal Sciences 101(Suppl. 3): 545-546. DOI: 10.1093/jas/skad281.641

386. P. Folchini, B. J. Duran, C. Rykaczewski, A. E. Crist, M. Saad, A. C. Carranza-Martin, J. C. L. Motta, D. F. Mollenkopf, M. L. Mussard, A. Garcia-Guerra. “Induction of endometritis after pregnancy loss during the second month of gestation in cattle and its effect on the fate of the corpus luteum” Proceedings of the 56th Annual Meeting of the Society for the Study of Reproduction 2023, July 11th – 14th, Ottawa, Canada, Page 386.

380. F. Incarnato, A. E. Crist, J. C. L. Motta, C. B. Hayden, A. Garcia-Guerra. “Assessment of circulating anti-Müllerian hormone during pregnancy in beef cattle” Proceedings of the 56th Annual Meeting of the Society for the Study of Reproduction, July 11th – 14th, 2023, Ottawa, Canada, Page 380.

2023. C. L. Motta, R. V. Sala, C. Hayden, D. C. Pereira, P. J. Ross, J.F. Moreno, A. Garcia-Guerra. 2023. “Ovarian stimulation with FSH increases in vitro embryo production in high AMH heifers in a dose-dependent manner”. Animal – science proceedings 14(3): 492. DOI: 10.1016/j.anscip.2023.03.099

2023. Carranza-Martin, C. Furnus, A. García-Guerra, A. Relling. 2023. “Effects of polyunsaturated fatty acids supplementation in resolvin D1 concentration in plasma and follicular fluid in ewes”. Journal of Animal Sciences 101(Suppl. 2): 332-333. DOI: 10.1093/jas/skad341.378

2023. B. Hayden, J. Motta, N. Bello, R. Sala, V. Absalon-Medina, P. Ross, J. Moreno, A. Garcia-Guerra. 2023. “Assessment of methods to synchronise follicle wave emergence in pregnant heifers”. Reproduction Fertility and Development 35(2):204-205. DOI: 10.1071/RDv35n2Ab153.

Hutchings DW, Elder D, Tsang PCW 2024. Effects of Phytoestrogens on Progesterone Biosynthesis in Dairy Cows. Poster; ASAS-CSAS-WSASAS Annual Meeting July 2024, Calgary, Canada.

Maucieri AM, Townson DH 2024. Influence of O-GlcNAcylation on KGN cell function and metabolism. Endocrine and Metabolic Science (In Revision)

Alward KJ, Cockrum RR, Ealy AD. Associations of antral follicle count with fertility in cattle: A review. JDS Commun. 2023 Jan 31;4(2):132-137. doi: 10.3168/jdsc.2022-0283.

Nix J, Marrella MA, Oliver MA, Rhoads M, Ealy AD, Biase FH. Cleavage kinetics is a better indicator of embryonic developmental competency than brilliant cresyl blue staining of oocytes. Anim Reprod Sci. 2023 Jan;248:107174. doi: 10.1016/j.anireprosci.2022.107174.

Nix JL, Schettini GP, Speckhart SL, Ealy AD, Biase FH. Ablation of OCT4 function in cattle embryos by double electroporation of CRISPR-Cas for DNA and RNA targeting (CRISPR-DART). PNAS Nexus. 2023 Oct 20;2(11):pgad343. doi: 10.1093/pnasnexus/pgad343.

Rhoads ML. Review: Reproductive consequences of whole-body adaptations of dairy cattle to heat stress. Animal. 2023 May;17 Suppl 1:100847. doi: 10.1016/j.animal.2023.100847.

Soffa, D.R., J.W. Stewart, E.D. Pack, A.G. Arneson, R. De Vita, J.W. Knight, D.W. Fausnacht, R.P. Rhoads, S.G. Clark, D.G. Schmale and M.L. Rhoads. 2023. Short-term consumption of the mycotoxin zearalenone by pubertal gilts causes persistent changes in the histoarchitecture of reproductive tissues. J Anim Sci. 2023 Jan 3;101:skac421. doi: 10.1093/jas/skac421. PMID: 36574505.

Speckhart SL, Oliver MA, Ealy AD. Developmental Hurdles That Can Compromise Pregnancy during the First Month of Gestation in Cattle. Animals (Basel). 2023 May 25;13(11):1760. doi: 10.3390/ani13111760.

Speckhart SL, Wooldridge LK, Ealy AD.  An updated protocol for in vitro bovine embryo production. STAR Protoc. 2023 Mar 17;4(1):101924. doi: 10.1016/j.xpro.2022.101924.

Arneson AG, Stewart JW, Byrd MH, Perry GA, Rhoads ML. Plasma γ-Aminobutyric Acid (GABA) Concentrations in Lactating Holstein Cows during Thermoneutral and Heat Stress Conditions and Their Relationships with Circulating Glucose, Insulin and Progesterone Levels. Vet Sci. 2024 Mar 21;11(3):137. doi: 10.3390/vetsci11030137.

Keane JA, Ealy AD. An Overview of Reactive Oxygen Species Damage Occurring during In Vitro Bovine Oocyte and Embryo Development and the Efficacy of Antioxidant Use to Limit These Adverse Effects. Animals (Basel). 2024 Jan 21;14(2):330. doi: 10.3390/ani14020330.

McKinley E, Speckhart SL, Keane JA, Oliver MA, Rhoads ML, Edwards JL, Biase FH, Ealy AD.  Influences of Supplementing Selective Members of the Interleukin-6 Cytokine Family on Bovine Oocyte Competency. Animals (Basel). 2023 Dec 21;14(1):44. doi: 10.3390/ani14010044.

Oliver MA, Speckhart SL, Ealy AD. Interleukin-6 Family Members Improve Pre-Implantation Bovine Blastocyst Composition. 2023 American Association of Animal Science Meeting, Albuquerque, NM.

Oliver MA, Speckhart SL, Edwards JL, Ealy AD. Human Recombinant Interleukin-6 has Cryoprotective Properties in Preimplantation Bovine Embryos. 2023 Society for the Study of Reproduction Annual Meeting, Ottawa, ON, CA.  

McKinley E, Speckhart SL, Keane JA, Oliver MA, Ealy AD. Supplementing IL6, IL11, and LIF to improve culture bovine oocyte competency. 2023 Society for the Study of Reproduction Annual Meeting, Ottawa, ON, CA.  

Nix JL, Speckhart SL, Schettini GP, Ealy AD, Biase FH. Disruption of OCT4 in cattle embryos by electroporation of CRISPR-Cas ribonucleoproteins into zygotes for DNA and RNA targeting (CRISPR-DART). 2023 Society for the Study of Reproduction Annual Meeting, Ottawa, ON, CA.  

Speckhart SL, Oliver MA, Keane JA, Dias NW, Mercadante VRG, Biase FH, Ealy AD. Interleukin-6 control of conceptus elongation and survival is multifaceted in the cow. 2023 Society for the Study of Reproduction Annual Meeting, Ottawa, ON, CA.  

Keane JA, Wooldridge LK, Speckhart SL, Nix JL, Biase FH, Ealy AD. Modifications in media composition supports increased blastocyst development and inner cell mass numbers of in vitro produced bovine embryos. 2023 Society for the Study of Reproduction Annual Meeting, Ottawa, ON, CA.  

Habeeb, H. M. H., Kleditz, L., Hazzard, T. M., Bishop, C. V., Stormshak, F., Kutzler, M. A. (2023). Ovine endometrial estrogen receptor expression is altered following PG-600 administration. Veterinary Medicine and Science. 9(3). DOI:10.1002/vms3.1119

Kim, S.-Y., Xu, J., Mayerhofer, A., Bishop, C. V. (2023). Editorial: Small molecules and peptides in paracrine/autocrine regulation of ovarian folliculogenesis. Frontiers in Endocrinology, 14. DOI:10.3389/fendo.2023.1168701

Slayden, O., Bishop, C. V. The Primate Uterus: Cyclic Changes Encyclopedia of Reproduction (3rd Edition). Accepted for Publication November 2023. Elsevier.

Bishop, C., Stricklin, O., Fox, A., Harrington, S., Estill, C., Zelinski, M., Ting, A. “Ovarian activity after autologous cryopreservation and transplantation in ewes.” Poster Presentation, 56th Annual Meeting, Society for the Study of Reproduction, July 11-14 2023, Ottawa, Canada.

Carr, S.N., B.R. Crites, H. Shinde and P.J. Bridges.  2023.  Transcriptomic changes in response to form of selenium on the interferon-tau signaling mechanism in caruncular tissue of beef heifers at maternal recognition of pregnancy.  International Journal of Molecular Sciences.  24:17327. doi:10.3390/ijms242417327.

S.N. Carr, B.R. Crites and P.J. Bridges.  2023.  Effects of form of selenium on serum metabolites and iodothyronine deiodinases during early gestation in beef heifers.  Special Issue, Proc. of the 56th annual meeting of the Soc. for the Study of Reproduction.

Rial, C., Hussain, I., Hoff, R., Tompkins, S., Erickson, D., Branen, J. and Giordano, J.O.* 2024. Development and evaluation of a lateral flow-based portable optical system for determination of the pregnancy status of dairy cows. J. Dairy Sci.  Online ahead of print. https://doi.org/10.3168/jds.2024-24899

Laplacette A. L., Rial C., Magaña Baños G. S., García Escalera J. S., Torres S., Kerwin A., and Giordano J. O.* 2024. Effect of a targeted reproductive management program based on automated detection of estrus during the voluntary waiting period on reproductive performance of lactating dairy cows. Theriogenology 225:130-141. https://10.1016/j.theriogenology.2024.05.030     

Sitko E. M., Laplacette A. L., Duhatschek D., Rial C., Perez M. M., Tompkins S., Kerwin A. L., Domingues R. R., Wiltbank  M. C. and Giordano J. O.* 2023. Ovarian function and endocrine phenotypes of lactating dairy cows during the estrous cycle are associated with genomic-enhanced predictions of fertility potential. J. Dairy Sci. 107:7352-7370. https://10.3168/jds.2023-24378  

Sitko E. M., Laplacette A. L., Duhatschek D., Rial C., Perez M. M., Tompkins S., Kerwin A. L.  and Giordano J. O.* 2023. Reproductive physiological outcomes of lactating dairy cows with different genomic merit for fertility: biomarkers, uterine health, endocrine status, estrus features, and response to synchronization of estrus and ovulation. J. Dairy Sci. https://10.3168/jds.2023-24376 Online ahead of print.

Rial C., and J. O. Giordano*. 2023. Combining reproductive outcomes predictors and automated estrus alerts recorded during the voluntary waiting period identified subgroups of cows with different reproductive performance potential. J. Dairy Sci. 107:7299-7316. https://10.3168/jds.2023-24309

Laplacette A. L., Rial C., Duhastchek D., Perez M.M., Stangaferro M. L., Thomas M. J. and Giordano J. O.* 2023. Effect of delaying induction of ovulation on ovarian function of lactating dairy cows synchronized with Double-Ovsynch and inseminated after detected estrus or timed AI. J. Dairy Sci. 106 (E-Suppl. 1).

Rial C., Stangaferro M. L., Thomas M. J. and Giordano J. O.* 2023. Association between vaginal discharge scores with rumination time, activity time, a health index score, and milk yield in lactating dairy cows. J. Dairy Sci. 106 (E-Suppl. 1).

Rial C., Hussain I., Erickson D., Branen J. and Giordano J. O.* 2023. Development and demonstration of lateral-flow immunoassays for determination of the pregnancy and ovarian physiological status of cows. J. Dairy Sci. 106 (E-Suppl. 1).

Theses/Dissertations

IMMUNOLOGICAL AND ENDOMETRIAL CHANGES OF EARLY PREGNANCY IN DAIRY CATTLE. 2023. Doctoral Dissertation in Integrative and Biomedical Physiology. Maria Isabel da Silva, Pennsylvania State University.

DeCastro, LN (2024) Evaluation of the effects of O-GlcNAcylation and Hypoxia on Estradiol Production by Bovine Granulosa Cells. URI MS Thesis

Endya McKinley (2023). Investigating the supplementation of IL-6, IL-11, & LIF at in vitro maturation to improve oocyte competency (MS Thesis).

Savannah L. Speckhart (2023).  Interleukin-6 and its Contribution to Embryogenesis in Cattle (PhD Dissertation).

Carr, Sarah Nancy, "SELENIUM FORM-INDUCED CHANGES IN THE EARLY LUTEAL PHASE CORPUS LUTEUM, THE BLOOD, AND THE ENDOMETRIUM DURING EARLY GESTATION IN BEEF COWS" (2023). Theses and Dissertations--Animal and Food Sciences. 138. https://uknowledge.uky.edu/animalsci_etds/138