Brief Summary of Minutes of Annual Meeting

Outputs from the group during this reporting period

1. 95 peer-reviewed manuscripts
2. 39 published abstracts
3. 22 invited oral presentations
4. 32 thesis/dissertations
5. 4 book chapters

Activities

Objective 1: Elucidate fundamental cellular, physiological, endocrine, and behavioral mechanisms that regulate gamete development and quality and enhance the management of reproductive function leading to the development of translational reproductive biotechnologies.

1. We have established stable ovine embryonic stem cell lines and demonstrated their plasticity across the pluripotency spectrum, providing a valuable platform for investigating ruminant stem cell biology and advancing livestock biotechnology. South Dakota
2. Studies to evaluate differences in in vivo sperm viability in cattle have been ongoing using a heterospermic artificial insemination approach. Kansas
3. We hypothesized that interferon-tau (IFNT) could be used to diagnose pregnancy in lactating cows. Results of our work demonstrate that detection of IFNT in cervical fluid via ELISA provides an accurate indication of pregnancy in lactating dairy cows.
4. Using RNA-Seq data from pooled early and expanded blastocysts categorized by quality grade, three mRNA transcripts were found to be up-regulated in grade 3 vs grade 1 embryos. These transcripts in addition to other embryo evaluations may identify the best candidates for embryo transfer. Colorado
5. We have developed a protocol for producing uterine organoids. Preliminary data from this work suggests that organoids derived from follicular vs. luteal stage tissue behave differently in culture. Montana
6. Using single-cell RNA-seq we have assessed the roles of Pramel1 and Pramex1 in germ cell differentiation and meiotic initiation in mice using knockout approaches. Collectively, results from this work demonstrate that Pramel1 and Pramex1 regulate pre-meiotic programs in germ cells during postnatal testis development. Pennsylvania
7. We have developed a sheep knockout model to study the role of follicle stimulating hormone (FSH) in gametogenesis. To date, four FSHβ knockout lambs have been born (3 male and 1 female). Follow-up evaluation of the males indicates that FSHβ knockout does not affect growth or sexual behaviors but does impact scrotal volume and causes oligospermia with increased frequency of sperm defects. California
8. Work to elucidate the roles of follicle stimulating hormone (FSH) in preantral folliculogenesis is ongoing. Present results suggest that exposure to supraphysiological levels of FSH leads to increased apoptosis in preantral follicles when compared to physiological FSH. California
9. We developed a mouse model of phthalate exposure using dibutyl phthalate (DBP) in which CD-1 female mice are orally exposed to the parent compound DBP at human relevant doses. Our results provide evidence that pituitary hormone release, ovarian sensitivity to gonadotropins, and preimplantation embryo development are impaired following daily oral exposure to DBP. Arizona
10. Research to identify proteomic differences that may contribute to the formation of aggregates in sperm was performed. No differences in protein abundances were observed but potential differentially modified post-translational modifications were identified. Wisconsin
11. We hypothesized that follicle stimulating hormone (FSH) induces coordinated metabolic reprogramming in granulosa cells, thereby altering the composition of follicular fluid. Our findings indicate that FSH drives coordinated metabolic reprogramming in granulosa cells, promoting lipid utilization and intracellular metabolic processing. Wisconsin
12. We evaluated the effects of follicle stimulating hormone on cumulus cell function and identified transcriptomic changes that suggest that FSH stimulation shifts cumulus cell metabolism from basal nutrient maintenance toward enhanced substrate exchange, lipid mobilization, and oxidative metabolism. Wisconsin
13. We used a bottom-up LC-MS/MS proteomic approach to compare the proteome of fresh versus cryopreserved spermatozoa from five bulls with proven fertility records. Results provide a proteomic atlas of cryopreservation damage in bull spermatozoa and identify candidate protein biomarkers for assessing cryotolerance prior to freezing. Iowa

14. We evaluated which sperm biomarkers most reliably distinguish true capacitation from cryopreservation-induced damage in cattle, a critical distinction for interpreting post-thaw sperm quality in AI programs. Phosphatidylserine translocation, cholesterol spatial distribution, and acrosome reaction status were the most reliable markers for differentiating genuine capacitation events from cryodamage artifacts. Iowa
15. We compared INRA96 (milk-based) and OviPlus (egg yolk-based) extenders for ram sperm stored at 4°C over 96 hours using image-based flow cytometry (IBFC) and computer-assisted semen analysis (CASA). Findings from this work provide the first zinc signature-based assessment of ram semen extenders and demonstrate that egg yolk-based formulations better protect ram spermatozoa against premature capacitation and oxidative damage during extended cooled storage. Iowa
16. Using the porcine model, we evaluated 206 sperm parameters to determine which biomarkers best predict in vitro fertilization (IVF) cleavage success. Our results validate zinc signatures as actionable IVF outcome predictors and are being translated to bovine IVF systems, where preliminary trials with W4112 collaborators are underway. Iowa
17. We performed LC-MS lipidomics paired with image-based flow cytometry on porcine spermatozoa under conditions of non-capacitated, capacitated, and capacitated with 2.0 mM zinc. Results from this work demonstrate that zinc ions stabilize over one-third of capacitation-associated lipid changes, with direct implications for understanding membrane competency for fertilization across mammalian species. Iowa
18. Using the porcine model to establish proof-of-concept for anti-ferroptotic semen preservation strategies, we treated semen with ferroptosis inducers (Erastin, RSL-3) and inhibitors (Ferrostatin-1, NAC) during 17°C liquid storage. Ferrostatin-1 and NAC reduced nearly all stressed subpopulations below untreated baseline levels. This is the first report connecting the ferroptosis pathway to semen storage outcomes in any species. Iowa
19. We evaluated the impact of sorting and catch media formulations on sperm motility, capacitation state, and viability using boar semen. Results demonstrate that sorting stress manifests with a delayed onset during capacitation, and that catch media composition is critical for maintaining post-sort sperm quality. Iowa
20. Work to examine the impact of oral melatonin supplementation on fresh and post-thawed sperm traits as well as scrotal temperatures was performed. Melatonin supplementation from fall to winter had both acute and long-term impacts on sperm kinematic and flow cytometric parameters. Factors related to motility were improved without negatively affecting scrotal temperatures. Mississippi

Objective 2: Identify impacts of reproductive management, animal management, and stress on follicle recruitment, ovulation, corpus luteum function, and pregnancy.

21. We have segregated yearling heifers according to their ability to attain puberty earlier or later in their first breeding season and have discovered blood markers that predict puberty attainment. Florida

22. We have validated the use of sensor technology, based on accelerometers, to identify and quantify intensity of estrous behavior in beef cows. Florida
23. We have collaborated with international partners to fine-tune the use of blood markers to diagnose early pregnancy status in cattle. Florida
24. We have evaluated the effects of extending CIDR insertion from 7 to 8 or 9 days on follicular dynamics, estrous expression, and pregnancy per AI in beef cows. Results suggest that extended progesterone exposure alters follicular development and may reduce fertility in some animals, particularly those with smaller dominant follicles. Missouri
25. Efforts to determine the effect of melengestrol acetate (MGA) and CIDR treatments, together with corpus luteum Color blood-flow Doppler ultrasonography, on early re-insemination of beef heifers have been performed. Preliminary results suggest that resynchronization of heifers using a 7-d CIDR treatment, followed by Doppler ultrasonography at day 24 after the first AI, was more effective than the MGA treatment for early re-insemination of more heifers. Missouri
26. Our work using gene co-expression network analysis demonstrates that there are distinct differences in granulosa cell signaling networks between heifers classified as Early and Non-Cycling. Moreover, the consistent patterns of expression observed in both heifers and cows support the idea that reproductive phenotypes established before puberty persist into adulthood. Nebraska
27. Heifers with excess ovarian androstenedione (A4) production exhibit early and persistent hematologic changes consistent with low-grade systemic inflammation, and whole blood indices, particularly, red blood cell-associated parameters, which may serve as early biomarkers for identifying High A4 females prior to breeding. Nebraska
28. Integrating pubertal classification and anogenital distance measures has allowed us to generate a ROC curve that may serve as a threshold to predict females with greater reproductive longevity and greater fertility. Nebraska
29. We recently observed that luteal cells exhibit features resembling senescence. Work from our group indicates that transforming growth factor-β together with other signaling molecules produced by follicular cells may play a role in reprogramming granulosa cells to become senescent as the follicle develops and eventually become large luteal cells. Nebraska
30. We identified a novel follicle stimulating hormone receptor single nucleotide polymorphism (SNP). The SNP is not associated with excess androgen females but instead is associated with females with greater birthweight and reduced weaning weight. We are determining if it segregates to delayed puberty females. Nebraska
31. We have determined that cows with high androstenedione have an altered theca cell transcriptome with increased androgen production, cell cycle arrest, and fibrosis in ovaries, exhibiting a PCOS-like phenotype. Nebraska
32. Efforts to identify proteins targeted by pregnancy induced ISGylaton in the ovine corpus luteum (CL) indicate that pregnancy activated type I interferon pathways and ISGylation in the CL. Most importantly, ISGylation appears to be targeting cholesterol synthesis, which may serve to stabilize steroidogenesis for sustained production of progesterone from the CL during pregnancy. Colorado
33. Research has been performed to assess the transcriptional response of ovine gonadotropes to estradiol. Results suggest that transcription factors downstream of insulin signaling, particularly FOXO1, may play a role in GNRH receptor and luteinizing hormone subunit beta (LHB) transcription. Colorado
34. Efforts to enrich gonadotropes from dissociated sheep pituitary cells using magnetic cell separation have been ongoing. Use of the cell surface marker calcium-activated potassium channel subunit alpha-1 (KCNMA1) resulted in an approximately 5-fold enrichment of gonadotropes as compared to no antibody controls. Colorado
35. Work to identify which population of neurons in the brainstem are activated during the preovulatory luteinizing hormone (LH) surge found that no population of catecholamine neuron in the ovine brainstem was activated during the LH surge. Instead, we found other (non- catecholamine) cells in the nucleus of the solitary tract and the dorsal lateral medulla were activated. Colorado
36. Gestational mineral requirements are well-studied in cattle, however, there is limited research on the effects of supranutritional mineral supplementation to calf development. We found that calves derived from dams administered a supraphysiologic dose of trace minerals during mid-gestation were heavier at birth and bulls derived from this treatment had sperm with greater progressive motility. Montana
37. Despite the knowledge that exercise is beneficial for overall health, how maternal exercise indirectly benefits the fetus and/or placenta is not well understood. An ongoing study seeks to determine the effects of low intensity exercise during gestation on maternal health and fetal-placental growth. Kentucky
38. We have investigated luteal responses to prostaglandin F₂α before and after maternal recognition of pregnancy and assessed endocrine and molecular changes associated with luteal maintenance. These studies supported a re-evaluation of classical paradigms regarding pregnancy maintenance, particularly the role of conceptus-derived interferon tau beyond its traditional function. Ohio
39. Research has been performed to assess the efficacy of supplemental progesterone as a strategy to enhance fetal and placental growth in sheep. Collectively, data from our work demonstrate that sustained, producer-friendly P4 supplementation via use of an intravaginal CIDR device can enhance fetal growth and drive organ-specific developmental changes. Pennsylvania
40. We have worked to characterize the relationship between reproductive microbiota and estradiol at ovulation in beef cattle and the relationship with fertility/conceptus growth. Minimal effects of estradiol were observed, but shifts in microbiota were observed across different gestational days and results suggest conceptus-derived factors may play a role. Texas
41. Efforts have been made to assess the relationship between feed efficiency and reproductive measures in beef heifers. Heifers were submitted to a feed efficiency trial and subsequently categorized as average, efficient or inefficient. No effect of feed efficiency category was observed for the reproductive traits measured. Idaho
42. Artificial insemination (AI), pregnancy detection, pelvic measurements and other reproductive technologies are used by less than 38% of the beef operations in the US. Extension efforts devoted to training ranchers in performing reproductive technologies have continued, including hosting two to three AI and ultrasound workshops annually. Idaho
43. We have performed research to assess the effects of microplastics on preantral bovine follicles. Microplastics demonstrated limited penetrative capacity in bovine preantral follicles during a 24-h in vitro exposure and appear to accumulate primarily within the follicular basement membrane. There were also no detectable changes in mitochondrial activity. California
44. We are using a novel approach to address fetal growth restriction due to placental insufficiency, which is to directly provide a balanced mixture of oxygen and glucose to the fetus. Our results indicate this approach can reverse impacts of fetal growth restriction. Arizona
45. Work has been performed to determine the impact of in utero exposure to environmentally relevant concentrations of atrazine on dopamine neurons in the ventral tegmental area (VTA) of brains of neonatal boars. Results indicate that in utero exposure to atrazine causes a depletion in tyrosine hydroxylase expressing neurons in the VTA of juvenile boars. Wyoming

46. Efforts to assess the effect of semen type on embryo development and pregnancy establishment after timed artificial insemination (TAI) or fresh embryo transfer (ET) in multiparous Holstein cows have been ongoing. Interactions between semen type and service type (i.e., TAI and ET) affected pregnancy rate and pregnancy loss. Wisconsin
47. We hypothesized that balancing essential amino acids (AAs) in the diet of lactating dairy cows would improve pregnancy outcomes and modulate uterine gene expression. Pregnancy rates did not differ between diets, but findings demonstrate that balancing AAs remodeled the uterine transcriptome across immune, metabolic, and reproductive gene networks. Wisconsin
48. We contributed sperm quality assessments (Image based flow cytometry zinc signatures, CASA motility) to a study of 44 half-sibling yearling beef bulls fed at moderate or high planes of nutrition for 114 days. Results demonstrate that overnutrition during the peri-pubertal period can induce metabolic dysregulation that impairs semen quality. Iowa
49. The effects of third-trimester melatonin supplementation on the growth, development, and behavior of Angus beef calves was evaluated. Melatonin decreased heart and abdominal girths without altering other growth metrics, confirming localized changes, but increased post-weaning behavioral reactivity, suggesting stress response alterations requiring further research. Mississippi

Objective 3: Determine mechanisms regulating normal embryo development, pregnancy establishment, and maintenance by exploring maternal and paternal factors; including genomics, immune responses, fetal programming, and conceptus/uterine signaling.

50. We have identified changes in gene expression and metabolite abundance in the uterine lumen during the second week of the estrous cycle. These changes may be associated with developing milestones of the embryo. Florida

51. We have discovered changes in gene expression and metabolite abundance in the uterine lumen, expression of molecular markers in blood cells (such as ISG15) and abundance of endocrine markers in the blood (such as PAGs) that are associated with pregnancy outcome to embryo transfer and ability of individual cows to retain pregnancy. Florida
52. We have tested the ability of uterine cells from cows to grow in culture and to respond to pregnancy and immune signals. We determined that cows with a lower ability to retain pregnancies have an exacerbated response to inflammatory signals (such as LPS). Florida

53. We previously reported that shearing fall-bred pregnant ewes on day 50 of gestation improves umbilical blood flow and fetal development. Our recent efforts indicate that shearing plays a role in placental gene expression, potentially contributing to the phenotypical effects previously reported. South Dakota
54. Lambs born from multiple pregnancies (i.e., twins and triplets) have generally lower birth weights than singletons. Recent work evaluating umbilical blood flow during mid-gestation indicates there is no difference between singletons and triplets. Twins might have a hemodynamic advantage, but this does not translate into similar lamb or placental weights. South Dakota
55. Efforts to evaluate effects of maternal diet on offspring performance suggest that the immune system of calves can be programmed prior to birth based on optimization of the dam’s protein intake. South Dakota
56. Research focused on determining the effects of administration of the chemokine ligand CXCL12 at the time of embryo transfer in sheep provide preliminary evidence that modulation of CXCL12 signaling represents a novel strategy to improve reproductive outcomes. New Mexico
57. Our work focused on understanding the role of CXCL12/CXCR4 in early placental functional development. We determined that CXCL12/CXCR4 signaling is not required for conceptus attachment but is important for coordinated development of vascular, immune, and metabolic pathways during early placentation in sheep. New Mexico
58. The insulin-like growth factor 2 receptor (IGF2R), a maternally expressed gene, is downregulated in overgrowth because of in-vitro embryo production (IVP). Results from work evaluating the effects of IVP on IGF signaling suggest that IVP affects IGF-PI3K-AKT-mTOR signaling in a tissue-specific manner and alters the relation of some proteins with fetal weight. We are designing a machine learning model to predict fetal weight in a sex-specific manner. Missouri
59. We have conducted research to investigate the enrichment of fertility-specific quantitative trait loci (QTL) within regulatory regions of the genes within the bovine placenta at single-cell resolution across different gestational time points. Washington
60. Disruptions in placental development can lead to pregnancy failure, making it essential to identify key conserved mechanisms that support pregnancy. We have identified cell types and gene expression dynamics in the post-implantation placenta of sheep and cattle to identify conserved mechanisms contributing to pregnancy success in ruminants. Washington
61. We have integrated single-cell transcriptomic atlases from mature bovine placenta and mid-gestation human placenta to characterize conserved and divergent placental cell populations and gene expression dynamics. Comparative analysis revealed strong conservation of immune cell types and transcriptional profiles between species, whereas trophoblast populations exhibited marked divergence in gene expression, consistent with known species-specific placental morphologies. Washington
62. Although morphological and histological differences exist in placentae across species, gene expression dynamics have not yet been compared at single-cell resolution. Work from our group reveals that in addition to morphological differences, there are also underlying molecular differences in cell types and gene expression dynamics between bovine and human placentae. Washington
63. While immune cell involvement in placentation and pregnancy loss has been studied more extensively in humans, limited information exists in cattle. Recent efforts have sought to comparatively analyze the gene expression profiles of immune cells in the bovine placenta at single-cell resolution. Washington
64. We hypothesized that epigenetic alterations observed in the splenic tissue of fetuses persistently infected with bovine viral diarrhea (BVDV) at gestational day 245 would persist in the postnatal period. Analysis revealed 8367 differentially methylated sites contained within genes associated with the immune and cardiac system, as well as hematopoiesis. Colorado
65. Ongoing research efforts aim to understand how the type of semen extender used impacts bull fertility by evaluating how differences in the metabolome, lipidome, and proteome relate to sperm fitness as quantified by flow cytometry. Montana
66. Efforts to assess the consequences of biological extremes in uterine crowding in pigs have been ongoing. Using surgical approaches to create either undercrowded or overcrowded pregnancies, we have found that uterine crowding drastically alters neonatal phenotypes. Kentucky
67. Maternal exposure to endocrine disrupting chemicals (EDCs) during gestation is concerning due to the potential for adverse developmental programming of the fetus. We have sought to assess the effects of gestational exposure of biosolids (a source of a complex mixture of EDCs) on placental function in ewes. Kentucky
68. Research from our group has focused on defining mechanisms of conceptus–maternal communication that regulate embryonic development, implantation timing, and pregnancy establishment in cattle. Using integrated in vivo and molecular approaches, our group investigated how variation in conceptus developmental capacity influences trophoblast differentiation, uterine receptivity, and placental function. Ohio
69. We have performed whole-genome bisulfite sequencing (WGBS) on embryos at different developmental stages from PRAMEY antibody-treated and rabbit IgG control groups. Integration of WGBS and RNA-seq datasets is underway to examine correlations between DNA methylation and gene expression and to test our hypothesis that PRAMEY regulates early embryo development through epigenetic mechanisms. Pennsylvania
70. Using a novel sheep model carrying a loss-of-function mutation in tissue non-specific alkaline phosphatase (TNSALP) we have sought to develop insights into placental phosphate availability. Collectively, our findings indicate that while TNSALP contributes to phosphate metabolism, its deficiency does not limit phosphate availability at the tissue level due to compensatory regulatory mechanisms. Pennsylvania
71. We have characterized the spatiotemporal expression of calcium and vitamin D regulatory molecules in bovine tissues collected from Day 46 to 136 of gestation. Our results demonstrate dynamic, tissue-specific regulation of calcium- and vitamin D–related pathways at the maternal–fetal interface. Pennsylvania
72. Work has been performed to characterize temporal changes in phosphate abundance, TNSALP activity (tissue non-specific alkaline phosphatase), and the expression and localization of TNSALP in bovine fetal fluids and utero-placental tissues during early- to mid-gestation. Results demonstrate dynamic, gestation-dependent regulation of TNSALP and support a role for this enzyme in modulating phosphate availability at the maternal–fetal interface in cattle. Pennsylvania
73. We have investigated the temporal changes in phosphate concentrations and the potential role of fibroblast growth factor-23 (FGF23) and its co-receptor klotho (KL) in cattle. Results from this work support the presence of localized and gestation-dependent regulatory mechanisms controlling phosphate availability and suggest that FGF23 and KL signaling may contribute to regulation of phosphate dynamics during pregnancy. Pennsylvania
74. Our group performed work to localize immune markers are the bovine utero-placental interface during early gestation. This study is one of the first to identify cell-type specific protein localization at bovine implantation sites on gestational days 20, 26, and 32 for ISG15 and TLR4. Texas
75. We have conducted studies aimed at exploring mechanisms by which cotyledonary villi invade caruncles to form placentomes in sheep. The work has identified differentially expressed genes (DEGs) that are upregulated in caruncles during the initial invasion of cotyledonary villus invasion. Maryland
76. We are investigating how early placentomes develop in cows. The work has identified a novel population of trophoblast that lines the caruncular crypts of placentomes in cows. Maryland
77. Utilizing single-cell sequencing, we have characterized the transcriptomic profiles of fetal peripheral blood mononuclear cells (PBMCs) from control and placental insufficiency-induced fetal growth restricted (PI-FGR) sheep at the single-cell level. Our data indicate that the shifted PBMC populations in PI-FGR fetuses may contribute to fetal endothelial dysfunction observed in PI-FGR fetal sheep through dysregulated PBMCs-endothelial cell interactions. Arizona
78. We hypothesized that betaine supplementation would alleviate the negative effects of maternal nutrient restriction in sheep. Overall, maternal nutrient restriction altered fetal muscle development and liver antioxidant capacity, with betaine supplementation providing partial, tissue specific responses, highlighting the need for further study of betaine as a targeted nutritional intervention during maternal nutrient restriction. Connecticut
79. Work from our group also evaluated the effects of maternal nutrient restriction and betaine supplementation on fetal muscle growth in sheep. Findings from this work demonstrate that maternal nutrient restriction alters fetal muscle cellularity in a muscle specific manner. While maternal betaine supplementation did not fully reverse the effects of nutrient restriction, it could potentially play a modulatory role. Connecticut
80. Efforts to determine the role of the membrane associated progesterone receptor, PGRMC1, in bovine embryo development have been ongoing. Using a CRISR/Cas9 approach we have determined that zygotic ablation of PGRMC1 impairs bovine embryo development in vitro and alters cell allocation and differentiation in the resulting blastocysts. Wyoming
81. Experiments were performed to assess the role of casein kinase 1 delta (CK1δ) and epsilon (CK1ε) in regulating the expression of the long non-coding RNA rhabdomyosarcoma 2-associated transcript (Rmst) in the murine female reproductive tract. The observed pattern of Rmst expression in the early postnatal uterus and oviduct suggests that it may regulate development of these tissues, and that it is molecularly ordered downstream of CK1δ/ε Wyoming
82. Prior work from our group demonstrated that conditional ablation of progesterone receptor membrane component-1 (Pgrmc1) and/or Pgrmc2 or conditional overexpression of Pgrmc1 resulted in subfertility that progressed to premature reproductive senescence. Recent efforts have further identified disrupted PGRMC homeostasis in transient ovarian somatic cells (peri-ovulatory granulosa and large luteal cells) as a driver of accelerated premature ovarian insufficiency (POI) and ovarian aging through the demise of the primordial follicle pool and deterioration of the somatic compartment. Wyoming
83. We performed fate-mapping studies to demonstrate the transition of simple cuboid and secretory glandular epithelial cells into barrier functioning luminal epithelial cells during uterine tissue regeneration that follows parturition in mice. Wyoming
84. Interferon-stimulated gene-15 is an evolutionarily conserved embryo-induced protein expressed in female reproductive and placental tissues in all mammalian species studied to date; yet its exact functions remain elusive. Work from our group supports a dual role for embryo-induced uterine ISG15 in pregnancy where it likely functions intracellularly to modulate critical metabolic pathways tied to energy utilization, and extracellularly as a cytokine that regulates uterine natural killer cell activity. Wyoming
85. Using a lentiviral-based approach, we sought to determine if progesterone receptor membrane component-1 (PGRMC1) influenced human endometrial cancer cell (Ishikawa) and xenograft tumor growth, angiogenesis and immune cell recruitment in immunocompromised mice. Collectively, our findings indicate that PGRMC1 is a novel target to prevent growth and progression of endometrial and breast cancer. Wyoming
86. We have evaluated the effect of reduced concentrations of antioxidant-related essential amino acids on embryo development and reactive oxygen species abundance under normal, and heat shock (HS) conditions in vitro. Limiting antioxidant-related essential amino acids may enhance the embryo’s resilience to heat-induced stress, potentially by modulating metabolic or stress-response pathways during early development. Wisconsin
87. We have investigated the impact of epidermal growth factor receptor (EGFR) signaling and pyruvate supplementation on cumulus cell metabolic programming and oocyte lipid accumulation during in-vitro maturation. Our findings demonstrate that EGFR signaling and extracellular pyruvate concentrations modulate cumulus cell metabolism, influencing glucose transport, glycolytic flux, and fatty acid oxidation. Wisconsin

Publications

Abdelhady, A. W., S. Winuthayanon, M. S. Ortega, A. M. Kelleher, and T. E. Spencer. 2026.

Directed differentiation of bovine trophoblast stem cells: A useful in vitro model for placenta

development. Proc Natl Acad Sci USA. 123:e2600783123.

https://doi.org/10.1073/pnas.2600783123

Absalon-Medina, V. A., R. V. Sala, D. C. Pereira, V. C. Fricke, I. Devkota, Z. L. Bonomo, D. M. Fuego, M. McDonald, J. M. Sánchez, M. B. Rabaglino, A. Matsakas, A. Vourekas, X. Fu, R. M.

Rivera, P. Lonergan, P. J. Ross, and C. A. Simintiras. 2025. Amniotic fluid metabolic biomarkers of fetal physiology and pregnancy success. 2025. Biol. Reprod. 114:998-1017. https://doi:10.1093/biolre/ioaf236

Andrade, J. P. N., R. R. Domingues, V. E. Gomez-León, G. Madureira, L. C. Sartori, G. F. Grillo, M. Fosado, R. V. Sala, and M. C. Wiltbank. 2025. Delayed embryonic attachment, circulating pregnancy-associated glycoproteins, and pregnancy loss in heifer embryo recipients. Theriogenology. 247:117561. https://doi.org/10.1016/j.theriogenology.2025.117561

Ashley R. L., M. Mullins, S. Salopek, K. Saucedo, E. Alvarez, M. Hunt, S. Baez, N. Kronlein, K. Valdez, L. Butler, and K. Ropp. 2026. CXCL12/CXCR4 signaling coordinates trophoblast, immune, vascular, and metabolic programming during early placentation in sheep. Biol. Reprod. (accepted).

Bennett, A., C. C. Rocha, A. Waheed, F. S. Mesquita, T. S. Maia, M. L Haimon, Q. A. Hoorn,

2025. Sagheer, C. J. Cuellar, H. Sultana, O. A. Ojeda-Rojas, R. L. Krisher, M. Rubessa, K. G. Pohler, P. J Hansen, P. Moriel, R. C. Chebel, and M. Binelli. 2025. Assessment and Prediction of Embryo Survival in Fertility-Classified Beef Cows. Theriogenology. 250:117560

https://doi.org/10.1016/j.theriogenology.2025.117560

Binelli, M., M. C Lopez-Duarte, A. Gonella-Diaza, F. A. C. C. Silva, G. Pugliesi, T. Martins, and C. C. Rocha. 2025. Effectors and predictors of conceptus survival in cattle: What is next? Domest. Anim. Endocrinol. 92:106939.https://doi.org/10.1016/j.domaniend.2025.106939

Bishop, J. V., A. Guzeloglu, T. Scheller, J. J. Docheff, C. L. Gonzalez-Berrios, H. Van Campen,

2025. M. Nett, A. L. Zezeski, T. W. Geary, W. W. Thatcher, and T. R. Hansen. 2025. Early

identification of bovine pregnancy status and embryonic mortality†. Biol. Reprod.

112(5):981–995. https://doi.org/10.1093/biolre/ioaf066

Botigelli R. C., R. B. Arcanjo, C. Guiltinan, J. M. Smith, A. J. Morton, and A. C. Denicol .2026. Low oxygen environment alters transcripts related to energy metabolism without altering the pluripotency core of bovine embryonic stem cells. Biol. Open. 2026 Apr 28:062352. https://doi.org/10.1242/bio.062352

Brenner, M. A., R. S. Marques, C. J. Posbergh, A. L. Zezeski, T. W. Geary, and S. R. McCoski. 2025. Maternal injectable mineral administration effects on calf growth and reproductive parameters. Animals (Basel) 15(3):330. https://doi.org/10.3390/ani15030330

Brown, W., M. Kuzniar, S. Retherford, B. Mitrisin, A. Ingols, J.-K. Tena, S. C. Sucheta, and J. Block. 2025. Effects of administration of a non-specific immunostimulant prior to mating on pregnancy establishment and fetal and placental development in mice. Biochem Biophys Rep. 44:102349. https://doi.org/10.1016/j.bbrep.2025.102349

Chasi, B., J. P. Andrade, P. L. J. Monteiro, B. Castro, E. Moreno, E. Anta, N. N. Ali, J. B.

Cole, M. C. Wiltbank, F. Peñagaricano, A. Z. Balboula, and M. S. Ortega. 2025. Association

of fertility traits with markers of oocyte competence in dairy cattle. J Dairy Sci. 108, 12814-12823. https://doi.org/10.3168/jds.2025-26726

Chasi, B., M. M. Herlihy, M. Schmitt, J. B. Cole, A. Z. Balboula, M. C. Wiltbank, F.

Peñagaricano, and M. S. Ortega. 2026. Association of fertility traits with embryo

development and pregnancy establishment. Journal of Dairy Science. 109:5773–5785.

http://doi.org/10.3168/jds.2025-27730

Chenchar, A., V. Mattos Pereira, A. Apte, B. D. Lyttle, A. E. Vaughn, T. Lehmann, J. R. Bardill, C. Zgheib, K. W. Liechty, K. C. S. Roballo, Z. Zhang, B. M. Alexander, and S. Nair. 2025. Targeting Cathepsin K to accelerate diabetic wound healing. ACS Pharmacol. Transl. Sci. 8(7):2258-2269. https://doi.org/10.1021/acsptsci.5c00295

Clemens, M. S. and C. Stenhouse. 2026. Significant variation among gilts may limit the ability

to detect sex differences during the pre-implantation period. Front. Vet. Sci.13:1829423. https://doi.org/10.3389/fvets.2026.1829423

Clemente L., C. Zhou, K. Chaiyakul, J. H. Adams, J. Jacobson, J. L. Austin, D. S Boeldt., I. M. Ong, and I.M. Bird. 2025. TNF but not VEGF induces secretion of multiple chemokines and cytokines by uterine artery endothelial cells - potential implications for preeclampsia. J. Mol. Endocrinol. 75(2):e250008. https://doi.org10.1530/JME-25-0008

Denicol A. C., and R. C. Botigelli. 2025. Dissecting the development of the early lineages and primordial germ cells in the bovine embryo. Anim. Reprod. 22(3):e20250098.

https://doi.org/10.1590/1984-3143-ar2025-0098

Domingues, R. R., J. Fox, E. Day, M. C. Wiltbank, and L. L. Hernandez. 2025. Uterotropic response to the antidepressant fluoxetine (Prozac) is dependent on a functional serotonin transporter and circulating serotonin. Sci. Rep. 15:43555. https://doi.org/10.1038/s41598-025-27477-w

Chen, Z., F. J. Liu, Y. X. Cheng, P. F. Li, L. Zhang, M. R. Riggs, W.-S. Liu, and Z. Zhu. 2026. miR-374b-5p suppresses melanoma progression by targeting VEGFC to inhibit MAPK signaling and reprogram the tumor microenvironment. Int. J. Mol. Sci. 27(6): 2854. https://doi.org/10.3390/ijms27062854

Ehresmann, L. X., F. P. J. da-Silva-Junior, H. Baroody, G. D. Dotts, E. G. Kidwell, P. M.

Bonacim, J. V. Bishop, A. Guzeloglu, T. R. Hansen, and R. R. Domingues. 2026.

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