{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T20:13:08Z","timestamp":1781899988318,"version":"3.54.5"},"reference-count":46,"publisher":"Oxford University Press (OUP)","issue":"6","license":[{"start":{"date-parts":[[2025,10,4]],"date-time":"2025-10-04T00:00:00Z","timestamp":1759536000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"DOI":"10.13039\/100005825","name":"USDA NIFA","doi-asserted-by":"publisher","award":["2023-67015-39785"],"award-info":[{"award-number":["2023-67015-39785"]}],"id":[{"id":"10.13039\/100005825","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,12,16]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Zinc is essential for the proper functioning of a variety of cells and tissues. Due to the limited understanding of the role of zinc in the bovine ovary, our objective was to investigate the role of zinc in bovine mural granulosa cell (mGC) viability and identify cell signaling pathways regulated by zinc in this cell type. Key indicators of cell behavior were measured after cultured bovine mGCs were incubated with increasing levels of a zinc chelator, TPEN. Cell viability decreased in a dose dependent manner, with a 91% loss of cell viability seen at 10\u00a0\u03bcM TPEN (p\u2009=\u20091.02 \u00d7 10\u22129), which corresponded to a 49% increase in caspase 3\/7 activity (p\u2009=\u20090.006) and an upregulation of p-YAP at Ser127 (p\u2009=\u20090.006). TRULI, a potent LATS1\/2 inhibitor, in combination with TPEN treatments, partially rescued the decrease in cell viability seen at 3.5\u00a0\u03bcM TPEN (p\u2009=\u20092.54 \u00d7 10\u22127) and the increase in caspase 3\/7 activity seen at 10\u00a0\u03bcM TPEN (p\u2009=\u20090.0003). Surprisingly, TPEN treatment for 6\u00a0h showed a dose dependent increase in relative fold change in expression of the known YAP gene targets, CCN1 (p\u2009=\u20093.85 \u00d7 10\u221210) and CCN2 (p\u2009=\u20091.95 \u00d7 10\u22127). While the mechanism by which increased Hippo activation mediates TPEN-stimulated apoptosis is still unknown, these results demonstrate that zinc is an essential micronutrient for bovine mGC viability, and that zinc deficiency promotes Hippo signaling leading to increased apoptosis in bovine mGCs.<\/jats:p>","DOI":"10.1093\/biolre\/ioaf222","type":"journal-article","created":{"date-parts":[[2025,10,2]],"date-time":"2025-10-02T11:59:33Z","timestamp":1759406373000},"page":"1404-1413","source":"Crossref","is-referenced-by-count":2,"title":["Zinc deficiency in bovine mural granulosa cells results in hippo pathway-mediated apoptosis"],"prefix":"10.1093","volume":"113","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5319-2040","authenticated-orcid":false,"given":"Allison","family":"Gabel","sequence":"first","affiliation":[{"name":"Department of Animal Science, Center for Reproductive Biology and Health, Pennsylvania State University, University Park , PA ,","place":["USA"]},{"name":"Huck Institutes of the Life Sciences, Pennsylvania State University, University Park , PA ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2875-7393","authenticated-orcid":false,"given":"Francisco","family":"Diaz","sequence":"additional","affiliation":[{"name":"Department of Animal Science, Center for Reproductive Biology and Health, Pennsylvania State University, University Park , PA ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2025,10,4]]},"reference":[{"key":"2025121605122217600_ref1","doi-asserted-by":"publisher","first-page":"1341","DOI":"10.1210\/endo-100-5-1341","article-title":"Androgen production by theca and granulosa isolated from proestrous rat follicles","volume":"100","author":"Fortune","year":"1977","journal-title":"Endocrinology"},{"key":"2025121605122217600_ref2","doi-asserted-by":"publisher","first-page":"51","DOI":"10.1016\/0303-7207(94)90278-X","article-title":"Follicular oestrogen synthesis: the \u2018two-cell, two-gonadotrophin\u2019 model revisited","volume":"100","author":"Hillier","year":"1994","journal-title":"Mol Cell Endocrinol"},{"key":"2025121605122217600_ref3","article-title":"Zinc dynamics regulate early ovarian follicle development","volume":"299","author":"Chen","year":"2023","journal-title":"J Biol Chem"},{"key":"2025121605122217600_ref4","doi-asserted-by":"publisher","first-page":"873","DOI":"10.1210\/en.2011-1599","article-title":"Zinc depletion causes multiple defects in ovarian function during the periovulatory period in mice","volume":"153","author":"Tian","year":"2012","journal-title":"Endocrinology"},{"key":"2025121605122217600_ref5","doi-asserted-by":"publisher","first-page":"115","DOI":"10.1186\/s13048-024-01442-z","article-title":"Zinc deficiency deteriorates ovarian follicle development and function by inhibiting mitochondrial function","volume":"17","author":"Liu","year":"2024","journal-title":"J Ovarian Res"},{"key":"2025121605122217600_ref6","doi-asserted-by":"publisher","first-page":"51","DOI":"10.1016\/j.ydbio.2013.01.015","article-title":"Acute dietary zinc deficiency before conception compromises oocyte epigenetic programming and disrupts embryonic development","volume":"376","author":"Tian","year":"2013","journal-title":"Dev Biol"},{"key":"2025121605122217600_ref7","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1095\/biolreprod.113.113910","article-title":"Preconception zinc deficiency disrupts postimplantation fetal and placental development in mice","volume":"90","author":"Tian","year":"2014","journal-title":"Biol Reprod"},{"key":"2025121605122217600_ref8","doi-asserted-by":"publisher","first-page":"695","DOI":"10.1017\/S1431927615000380","article-title":"Localization of the trace elements iron, zinc and selenium in relation to anatomical structures in bovine ovaries by X-ray fluorescence imaging","volume":"21","author":"Ceko","year":"2015","journal-title":"Microsc Microanal"},{"key":"2025121605122217600_ref9","doi-asserted-by":"publisher","first-page":"865","DOI":"10.1111\/rda.12385","article-title":"The importance of having zinc during in vitro maturation of cattle cumulus\u2013oocyte complex: role of cumulus cells","volume":"49","author":"Anchordoquy","year":"2014","journal-title":"Reprod Domest Anim"},{"key":"2025121605122217600_ref10","doi-asserted-by":"publisher","first-page":"679","DOI":"10.1530\/REP-19-0398","article-title":"Zinc supports transcription and improves meiotic competence of growing bovine oocytes","volume":"159","author":"Lodde","year":"2020","journal-title":"Reproduction"},{"key":"2025121605122217600_ref11","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3168\/jds.2016-11302","article-title":"Invited review: a systematic literature review and meta-analysis of mortality and culling in dairy cattle","volume":"100","author":"Compton","year":"2017","journal-title":"J Dairy Sci"},{"key":"2025121605122217600_ref12","doi-asserted-by":"publisher","first-page":"327","DOI":"10.1017\/S1751731116001348","article-title":"Associations between strain, herd size, age at first calving, culling reason and lifetime performance characteristics in Holstein-Friesian cows","volume":"11","author":"Adamczyk","year":"2017","journal-title":"Animal"},{"key":"2025121605122217600_ref13","doi-asserted-by":"publisher","first-page":"680","DOI":"10.1071\/AN20195","article-title":"Analysis of culling reasons and age at culling in Australian dairy cattle","volume":"61","author":"Workie","year":"2021","journal-title":"Anim Prod Sci"},{"key":"2025121605122217600_ref14","doi-asserted-by":"publisher","first-page":"28","DOI":"10.1093\/af\/vfz017","article-title":"Livestock animals to study infertility in women","volume":"9","author":"Abedal-Majed","year":"2019","journal-title":"Anim Front"},{"key":"2025121605122217600_ref15","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1016\/S0092-8674(02)00824-3","article-title":"Salvador promotes both cell cycle exit and apoptosis in drosophila and is mutated in human cancer cell lines","volume":"110","author":"Tapon","year":"2002","journal-title":"Cell"},{"key":"2025121605122217600_ref16","doi-asserted-by":"crossref","first-page":"5496","DOI":"10.1074\/jbc.M709037200","article-title":"Tumor suppressor LATS1 is a negative regulator of oncogene YAP","volume":"283","author":"Hao","year":"2008","journal-title":"J Biol Chem"},{"key":"2025121605122217600_ref17","doi-asserted-by":"publisher","first-page":"2747","DOI":"10.1101\/gad.1602907","article-title":"Inactivation of YAP oncoprotein by the hippo pathway is involved in cell contact inhibition and tissue growth control","volume":"21","author":"Zhao","year":"2007","journal-title":"Genes Dev"},{"key":"2025121605122217600_ref18","doi-asserted-by":"publisher","first-page":"1120","DOI":"10.1016\/j.cell.2007.07.019","article-title":"Elucidation of a universal size-control mechanism in drosophila and mammals","volume":"130","author":"Dong","year":"2007","journal-title":"Cell"},{"key":"2025121605122217600_ref19","doi-asserted-by":"crossref","first-page":"1962","DOI":"10.1101\/gad.1664408","article-title":"TEAD mediates YAP-dependent gene induction and growth control","volume":"22","author":"Zhao","year":"2008","journal-title":"Genes Dev"},{"key":"2025121605122217600_ref20","doi-asserted-by":"publisher","first-page":"267","DOI":"10.1016\/j.cell.2006.01.005","article-title":"From cell structure to transcription: hippo forges a new path","volume":"124","author":"Edgar","year":"2006","journal-title":"Cell"},{"key":"2025121605122217600_ref21","doi-asserted-by":"crossref","first-page":"1074","DOI":"10.1210\/endrev\/bnac013","article-title":"Hippo signaling in the ovary: emerging roles in development, fertility, and disease","volume":"43","author":"Clark","year":"2022","journal-title":"Endocr Rev"},{"key":"2025121605122217600_ref22","doi-asserted-by":"publisher","first-page":"10049","DOI":"10.1096\/fj.201900179RR","article-title":"Timely expression and activation of YAP1 in granulosa cells is essential for ovarian follicle development","volume":"33","author":"Lv","year":"2019","journal-title":"FASEB J"},{"key":"2025121605122217600_ref23","doi-asserted-by":"publisher","first-page":"113","DOI":"10.1186\/s12958-019-0552-1","article-title":"Ovulatory signals alter granulosa cell behavior through YAP1 signaling","volume":"17","author":"Sun","year":"2019","journal-title":"Reprod Biol Endocrinol"},{"key":"2025121605122217600_ref24","doi-asserted-by":"crossref","first-page":"1001","DOI":"10.1093\/biolre\/ioz139","article-title":"Yes-associated protein 1 is required for proliferation and function of bovine granulosa cells in vitro","volume":"101","author":"Plewes","year":"2019","journal-title":"Biol Reprod"},{"key":"2025121605122217600_ref25","doi-asserted-by":"publisher","first-page":"74","DOI":"10.1038\/s41556-021-00813-8","article-title":"The hippo pathway kinases LATS1 and LATS2 attenuate cellular responses to heavy metals through phosphorylating MTF1","volume":"24","author":"Han","year":"2022","journal-title":"Nat Cell Biol"},{"key":"2025121605122217600_ref26","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2\u2212\u0394\u0394CT method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"2025121605122217600_ref27","doi-asserted-by":"publisher","first-page":"2719","DOI":"10.1016\/S0021-9258(18)89421-2","article-title":"Cytosolic Ca2+ homeostasis in Ehrlich and Yoshida carcinomas. A new, membrane-permeant chelator of heavy metals reveals that these ascites tumor cell lines have normal cytosolic free Ca2+","volume":"260","author":"Arslan","year":"1985","journal-title":"J Biol Chem"},{"key":"2025121605122217600_ref28","doi-asserted-by":"publisher","first-page":"2330","DOI":"10.1002\/hlca.19670500817","article-title":"Pyridine derivatives as complexing agents. VIII. Preparation of a new quadridentate and a new hexadentate ligand","volume":"50","author":"Anderegg","year":"1967","journal-title":"Helv Chim Acta"},{"key":"2025121605122217600_ref29","doi-asserted-by":"crossref","first-page":"29","DOI":"10.4162\/nrp.2007.1.1.29","article-title":"Cellular Zn depletion by metal ion chelators (TPEN, DTPA and chelex resin) and its application to osteoblastic MC3T3-E1 cells","volume":"1","author":"Cho","year":"2007","journal-title":"Nutr Res Pract"},{"key":"2025121605122217600_ref30","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1186\/1471-2407-14-527","article-title":"Copper chelation selectively kills colon cancer cells through redox cycling and generation of reactive oxygen species","volume":"14","author":"Fatfat","year":"2014","journal-title":"BMC Cancer"},{"key":"2025121605122217600_ref31","doi-asserted-by":"publisher","first-page":"301","DOI":"10.1016\/j.jnutbio.2010.06.010","article-title":"The significance of copper chelators in clinical and experimental application","volume":"22","author":"Ding","year":"2011","journal-title":"J Nutr Biochem"},{"key":"2025121605122217600_ref32","doi-asserted-by":"crossref","first-page":"1500S","DOI":"10.1093\/jn\/130.5.1500S","article-title":"The role of zinc in growth and cell proliferation","volume":"130","author":"MacDonald","year":"2000","journal-title":"J Nutr"},{"key":"2025121605122217600_ref33","doi-asserted-by":"publisher","first-page":"14160","DOI":"10.3390\/ijms232214160","article-title":"FSH regulates YAP-TEAD transcriptional activity in bovine granulosa cells to allow the future dominant follicle to exert its augmented estrogenic capacity","volume":"23","author":"Andrade","year":"2022","journal-title":"Int J Mol Sci"},{"key":"2025121605122217600_ref34","doi-asserted-by":"publisher","first-page":"14","DOI":"10.1186\/s13048-023-01305-z","article-title":"Correlation between ovarian follicular development and hippo pathway in polycystic ovary syndrome","volume":"17","author":"Huang","year":"2024","journal-title":"J Ovarian Res"},{"key":"2025121605122217600_ref35","doi-asserted-by":"publisher","first-page":"72","DOI":"10.15252\/embr.201642681","article-title":"Osmotic stress-induced phosphorylation by NLK at Ser128 activates YAP","volume":"18","author":"Hong","year":"2017","journal-title":"EMBO Rep"},{"key":"2025121605122217600_ref36","doi-asserted-by":"crossref","first-page":"400","DOI":"10.3390\/v17030400","article-title":"Targeting latent HIV reservoirs: effectiveness of combination therapy with HDAC and PARP inhibitors","volume":"17","author":"Tibebe","year":"2025","journal-title":"Viruses"},{"key":"2025121605122217600_ref37","first-page":"dev201621","article-title":"Spatial cell fate manipulation of human pluripotent stem cells by controlling the microenvironment using photocurable hydrogel","volume":"151","author":"Wang","year":"2024","journal-title":"Dev Camb Engl"},{"key":"2025121605122217600_ref38","doi-asserted-by":"publisher","first-page":"3217","DOI":"10.1111\/jcmm.17879","article-title":"Histological characteristics of exercise-induced skeletal muscle remodelling","volume":"27","author":"Su","year":"2023","journal-title":"J Cell Mol Med"},{"key":"2025121605122217600_ref39","doi-asserted-by":"publisher","first-page":"169","DOI":"10.1677\/joe.0.1780169","article-title":"The CCN family: a new stimulus package","volume":"178","author":"Brigstock","year":"2003","journal-title":"J Endocrinol"},{"key":"2025121605122217600_ref40","doi-asserted-by":"publisher","first-page":"50","DOI":"10.1186\/1757-2215-7-50","article-title":"Changes in granulosa cells gene expression associated with growth, plateau and atretic phases in medium bovine follicles","volume":"7","author":"Douville","year":"2014","journal-title":"J Ovarian Res"},{"key":"2025121605122217600_ref41","doi-asserted-by":"publisher","first-page":"skab318","DOI":"10.1093\/jas\/skab318","article-title":"Effects of bone morphogenetic protein 4, gremlin, and connective tissue growth factor on estradiol and progesterone production by bovine granulosa cells","volume":"99","author":"Spicer","year":"2021","journal-title":"J Anim Sci"},{"key":"2025121605122217600_ref42","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1016\/j.mce.2015.11.009","article-title":"Growth differentiation factor 8 suppresses cell proliferation by up-regulating CTGF expression in human granulosa cells","volume":"422","author":"Chang","year":"2016","journal-title":"Mol Cell Endocrinol"},{"key":"2025121605122217600_ref43","doi-asserted-by":"publisher","first-page":"175","DOI":"10.1007\/s12011-019-1651-8","article-title":"Parenteral zinc supplementation increases pregnancy rates in beef cows","volume":"192","author":"Anchordoquy","year":"2019","journal-title":"Biol Trace Elem Res"},{"key":"2025121605122217600_ref44","doi-asserted-by":"publisher","first-page":"e99790","DOI":"10.1371\/journal.pone.0099790","article-title":"Serum and tissue zinc in epithelial malignancies: a meta-analysis","volume":"9","author":"Gumulec","year":"2014","journal-title":"PLoS One"},{"key":"2025121605122217600_ref45","doi-asserted-by":"publisher","first-page":"195","DOI":"10.1007\/s00404-020-05789-8","article-title":"Zinc distribution within breast cancer tissue of different intrinsic subtypes","volume":"303","author":"Rusch","year":"2021","journal-title":"Arch Gynecol Obstet"},{"key":"2025121605122217600_ref46","doi-asserted-by":"publisher","first-page":"e14","DOI":"10.1017\/erm.2015.12","article-title":"The hippo transducers TAZ and YAP in breast cancer: oncogenic activities and clinical implications","volume":"17","author":"Maugeri-Sacc\u00e0","year":"2015","journal-title":"Expert Rev Mol Med"}],"container-title":["Biology of Reproduction"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/biolreprod\/advance-article-pdf\/doi\/10.1093\/biolre\/ioaf222\/64493777\/ioaf222.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/biolreprod\/article-pdf\/113\/6\/1404\/64493777\/ioaf222.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/biolreprod\/article-pdf\/113\/6\/1404\/64493777\/ioaf222.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,16]],"date-time":"2025-12-16T10:12:32Z","timestamp":1765879952000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/biolreprod\/article\/113\/6\/1404\/8273750"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,4]]},"references-count":46,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2025,10,4]]},"published-print":{"date-parts":[[2025,12,16]]}},"URL":"https:\/\/doi.org\/10.1093\/biolre\/ioaf222","relation":{},"ISSN":["0006-3363","1529-7268"],"issn-type":[{"value":"0006-3363","type":"print"},{"value":"1529-7268","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2025,12]]},"published":{"date-parts":[[2025,10,4]]}}}