{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T18:56:00Z","timestamp":1791053760266,"version":"4.1.0"},"reference-count":51,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2021,5,10]],"date-time":"2021-05-10T00:00:00Z","timestamp":1620604800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Estrogen receptor beta (ER\u03b2) plays a critical role in granulosa cell (GC) functions. The existence of four human ER\u03b2 splice isoforms in the ovary suggests their differential implication in 17\u03b2-estradiol (E2) actions on GC apoptosis causing follicular atresia. In this study, we investigated whether E2 can regulate ER\u03b2 isoforms expression to fine tune its apoptotic activities in human GC. For this purpose, we measured by RT-qPCR the expression of ER\u03b2 isoforms in primary culture of human granulosa cells (hGCs) collected from patients undergoing in vitro fertilization, before and after E2 exposure. Besides, we assessed the potential role of ER\u03b2 isoforms on cell growth and apoptosis after their overexpression in a human GC line (HGrC1 cells). We confirmed that ER\u03b21, ER\u03b22, ER\u03b24, and ER\u03b25 isoform mRNAs were predominant over that of ER\u03b1 in hGCs, and found that E2 selectively regulates mRNA levels of ER\u03b24 and ER\u03b25 isoforms in these cells. In addition, we demonstrated that overexpression of ER\u03b21 and ER\u03b24 in HGrC1 cells increased cell apoptosis by 225% while ER\u03b25 or ER\u03b22 had no effect. Altogether, our study revealed that E2 may influence GC fate by specifically regulating the relative abundance of ER\u03b2 isoforms mRNA to modulate the balance between pro-apoptotic and non-apoptotic ER\u03b2 isoforms.<\/jats:p>","DOI":"10.3390\/ijms22095046","type":"journal-article","created":{"date-parts":[[2021,5,10]],"date-time":"2021-05-10T10:49:51Z","timestamp":1620643791000},"page":"5046","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Estradiol Regulates mRNA Levels of Estrogen Receptor Beta 4 and Beta 5 Isoforms and Modulates Human Granulosa Cell Apoptosis"],"prefix":"10.3390","volume":"22","author":[{"given":"Alice","family":"Pierre","sequence":"first","affiliation":[{"name":"BFA, UMR 8251, CNRS, ERL U1133, Inserm, Universit\u00e9 de Paris, F-75013 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne","family":"Mayeur","sequence":"additional","affiliation":[{"name":"Service de M\u00e9decine de la Reproduction et Pr\u00e9servation de la Fertilit\u00e9, H\u00f4pital Antoine B\u00e9cl\u00e8re, 92140 Clamart, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cl\u00e9mentine","family":"Marie","sequence":"additional","affiliation":[{"name":"BFA, UMR 8251, CNRS, ERL U1133, Inserm, Universit\u00e9 de Paris, F-75013 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victoria","family":"Cluzet","sequence":"additional","affiliation":[{"name":"BFA, UMR 8251, CNRS, ERL U1133, Inserm, Universit\u00e9 de Paris, F-75013 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathan","family":"Chauvin","sequence":"additional","affiliation":[{"name":"Lixoft, 8 rue de la Renaissance, B\u00e2timent D, 92160 Antony, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nelly","family":"Frydman","sequence":"additional","affiliation":[{"name":"Service de M\u00e9decine de la Reproduction et Pr\u00e9servation de la Fertilit\u00e9, H\u00f4pital Antoine B\u00e9cl\u00e8re, 92140 Clamart, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Grynberg","sequence":"additional","affiliation":[{"name":"BFA, UMR 8251, CNRS, ERL U1133, Inserm, Universit\u00e9 de Paris, F-75013 Paris, France"},{"name":"Service de M\u00e9decine de la Reproduction et Pr\u00e9servation de la Fertilit\u00e9, H\u00f4pital Antoine B\u00e9cl\u00e8re, 92140 Clamart, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joelle","family":"Cohen-Tannoudji","sequence":"additional","affiliation":[{"name":"BFA, UMR 8251, CNRS, ERL U1133, Inserm, Universit\u00e9 de Paris, F-75013 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4489-4304","authenticated-orcid":false,"given":"C\u00e9line J.","family":"Guigon","sequence":"additional","affiliation":[{"name":"BFA, UMR 8251, CNRS, ERL U1133, Inserm, Universit\u00e9 de Paris, F-75013 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8439-1453","authenticated-orcid":false,"given":"St\u00e9phanie","family":"Chauvin","sequence":"additional","affiliation":[{"name":"BFA, UMR 8251, CNRS, ERL U1133, Inserm, Universit\u00e9 de Paris, F-75013 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,5,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1262\/jrd.50.493","article-title":"Regulation mechanism of selective atresia in porcine follicles: Regulation of granulosa cell apoptosis during atresia","volume":"50","author":"Manabe","year":"2004","journal-title":"J. Reprod. Dev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1547","DOI":"10.1051\/rnd:19790916","article-title":"The human antral follicle: Functional correlates of growth and atresia","volume":"19","author":"McNatty","year":"1979","journal-title":"Ann. Biol. Anim. Bioch. Biophys."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/j.anireprosci.2004.04.031","article-title":"Follicular development: The role of the follicular microenvironment in selection of the dominant follicle","volume":"82\u201383","author":"Fortune","year":"2004","journal-title":"Anim. Reprod. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1038\/sj.cr.7290234","article-title":"Apoptosis in granulosa cells during follicular atresia: Relationship with steroids and insulin-like growth factors","volume":"14","author":"Yu","year":"2004","journal-title":"Cell Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1002\/ajp.1350230405","article-title":"Estradiol-induced follicular atresia in rhesus monkeys is not prevented by exogenous gonadotropins","volume":"23","author":"Hutz","year":"1991","journal-title":"Am. J. Primatol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1111\/j.1600-0684.1990.tb00463.x","article-title":"Role of estradiol in regulating ovarian follicular atresia in rhesus monkeys: A review","volume":"19","author":"Hutz","year":"1990","journal-title":"J. Med. Primatol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2204","DOI":"10.1210\/endo.133.5.8404672","article-title":"Estrogens inhibit and androgens enhance ovarian granulosa cell apoptosis","volume":"133","author":"Billig","year":"1993","journal-title":"Endocrinology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"5129","DOI":"10.1073\/pnas.0306720101","article-title":"Impact of isotype-selective estrogen receptor agonists on ovarian function","volume":"101","author":"Siebel","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1016\/j.mce.2011.09.026","article-title":"Estrogen dependent signaling in reproductive tissues\u2014A role for estrogen receptors and estrogen related receptors","volume":"348","author":"Gibson","year":"2012","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"681","DOI":"10.1016\/j.steroids.2004.06.004","article-title":"Endogenous estrogen receptor beta is transcriptionally active in primary ovarian cells from estrogen receptor knockout mice","volume":"69","author":"Mueller","year":"2004","journal-title":"Steroids"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"634","DOI":"10.1289\/EHP396","article-title":"Differential Activation of a Mouse Estrogen Receptor beta Isoform (mERbeta2) with Endocrine-Disrupting Chemicals (EDCs)","volume":"125","author":"Donoghue","year":"2017","journal-title":"Environ. Health Perspect"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"985","DOI":"10.1016\/S0039-128X(02)00047-8","article-title":"Human oestrogen receptors: Differential expression of ER alpha and beta and the identification of ER beta variants","volume":"67","author":"Scobie","year":"2002","journal-title":"Steroids"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1385\/ENDO:22:2:101","article-title":"Molecular assays to profile 10 estrogen receptor beta isoform mRNA copy numbers in ovary, breast, uterus, and bone tissues","volume":"22","author":"Poola","year":"2003","journal-title":"Endocrine"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1006\/bbrc.1998.8738","article-title":"Cloning and characterization of human estrogen receptor beta isoforms","volume":"247","author":"Moore","year":"1998","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"13162","DOI":"10.1073\/pnas.0605676103","article-title":"Estrogen receptor (ER)-beta isoforms: A key to understanding ER-beta signaling","volume":"103","author":"Leung","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1038\/nrc3093","article-title":"The different roles of ER subtypes in cancer biology and therapy","volume":"11","author":"Thomas","year":"2011","journal-title":"Nat. Rev. Cancer"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1530\/ERC-19-0291","article-title":"The ERbeta5 splice variant increases oestrogen responsiveness of ERalphapos Ishikawa cells","volume":"27","author":"Collins","year":"2020","journal-title":"Endocr. Relat. Cancer"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1677\/jme.0.0300013","article-title":"Putative functional characteristics of human estrogen receptor-beta isoforms","volume":"30","author":"Peng","year":"2003","journal-title":"J. Mol. Endocrinol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1385\/ENDO:27:3:227","article-title":"Estrogen receptors beta4 and beta5 are full length functionally distinct ERbeta isoforms: Cloning from human ovary and functional characterization","volume":"27","author":"Poola","year":"2005","journal-title":"Endocrine"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"3955","DOI":"10.1158\/0008-5472.CAN-06-3505","article-title":"Estrogen receptor beta2 negatively regulates the transactivation of estrogen receptor alpha in human breast cancer cells","volume":"67","author":"Zhao","year":"2007","journal-title":"Cancer Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1111\/j.1447-0578.2003.00043.x","article-title":"Correlations between steroids concentration in follicular fluid, pronuclear morphology and embryo qualities in in vitro fertilization","volume":"2","author":"Hasegawa","year":"2003","journal-title":"Reprod. Med. Biol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2344","DOI":"10.1210\/jc.2007-2342","article-title":"Concentration of anti-Mullerian hormone and inhibin-B in relation to steroids and age in follicular fluid from small antral human follicles","volume":"93","author":"Rosendahl","year":"2008","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2851","DOI":"10.1210\/en.2011-1810","article-title":"Establishment of a human nonluteinized granulosa cell line that transitions from the gonadotropin-independent to the gonadotropin-dependent status","volume":"153","author":"Kiyono","year":"2012","journal-title":"Endocrinology"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1016\/S0960-0760(00)00100-X","article-title":"Trafficking of nuclear receptors in living cells","volume":"74","author":"Hager","year":"2000","journal-title":"J. Steroid Biochem. Mol. Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1677\/JOE-07-0087","article-title":"Estrogen receptor \u03b21 exerts antitumoral effects on SK-OV-3 ovarian cancer cells","volume":"193","author":"Treeck","year":"2007","journal-title":"J. Endocrinol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"3760","DOI":"10.1210\/en.2016-1297","article-title":"Endocrine Response Phenotypes Are Altered by Charcoal-Stripped Serum Variability","volume":"157","author":"Sikora","year":"2016","journal-title":"Endocrinology"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1016\/j.lfs.2012.05.010","article-title":"Why two endothelins and two receptors for ovulation and luteal regulation?","volume":"91","author":"Ko","year":"2012","journal-title":"Life Sci."},{"key":"ref_28","first-page":"145","article-title":"Endothelins in regulating ovarian and oviductal function","volume":"3","author":"Bridges","year":"2011","journal-title":"Front. Biosci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1171","DOI":"10.1093\/humrep\/der066","article-title":"Expression pattern of endothelin system components and localization of smooth muscle cells in the human pre-ovulatory follicle","volume":"26","author":"Choi","year":"2011","journal-title":"Hum. Reprod."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1016\/j.lfs.2016.01.010","article-title":"Ovarian hormones modulate endothelin A and B receptor expression","volume":"159","author":"Gohar","year":"2016","journal-title":"Life Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"6248197","DOI":"10.1155\/2019\/6248197","article-title":"Enhanced Endothelin A and B Receptor Expression and Receptor-Mediated Vasoconstriction in Rat Mesenteric arteries after Lipopolysaccharide Challenge","volume":"2019","author":"Zhang","year":"2019","journal-title":"Mediat. Inflamm."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2172","DOI":"10.1111\/j.1582-4934.2009.00867.x","article-title":"Differential regulation of oestrogen receptor beta isoforms by 5\u2019 untranslated regions in cancer","volume":"14","author":"Smith","year":"2010","journal-title":"J. Cell. Mol. Med."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1042\/BJ20100373","article-title":"Expression of oestrogen receptor beta isoforms is regulated by transcriptional and post-transcriptional mechanisms","volume":"429","author":"Smith","year":"2010","journal-title":"Biochem. J."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/S0960-0760(01)00071-1","article-title":"The multiple untranslated first exons system of the human estrogen receptor beta (ER beta) gene","volume":"78","author":"Hirata","year":"2001","journal-title":"J. Steroid Biochem. Mol. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.mce.2013.06.023","article-title":"Differential expression of estrogen receptor beta isoforms in prostate cancer through interplay between transcriptional and translational regulation","volume":"376","author":"Lee","year":"2013","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1132","DOI":"10.1210\/me.2007-0022","article-title":"The micro-ribonucleic acid (miRNA) miR-206 targets the human estrogen recep-tor-alpha (ERalpha) and represses ERalpha messenger RNA and protein expression in breast cancer cell lines","volume":"21","author":"Adams","year":"2007","journal-title":"Mol. Endocrinol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"4850","DOI":"10.1093\/nar\/gkp500","article-title":"Estradiol-regulated microRNAs control estradiol response in breast cancer cells","volume":"37","author":"Wang","year":"2009","journal-title":"Nucleic Acids Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"4778","DOI":"10.1158\/0008-5472.CAN-09-4104","article-title":"Estrogen receptor \u03b21 expression is regulated by miR-92 in breast cancer","volume":"70","author":"Burns","year":"2010","journal-title":"Cancer Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"616","DOI":"10.1095\/biolreprod67.2.616","article-title":"Involvement of androgen receptor in 17beta-estradiol-induced cell proliferation in rat uterus","volume":"67","author":"Weihua","year":"2002","journal-title":"Biol. Reprod."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"3487","DOI":"10.1210\/en.2004-0212","article-title":"Expression of progesterone receptors A and B in the mouse ovary during the estrous cycle","volume":"145","author":"Gava","year":"2004","journal-title":"Endocrinology"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"439","DOI":"10.1146\/annurev.physiol.62.1.439","article-title":"The mechanism of action of thyroid hormones","volume":"62","author":"Zhang","year":"2000","journal-title":"Annu. Rev. Physiol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1016\/0092-8674(88)90194-8","article-title":"The thyroid hormone receptor binds with opposite transcriptional effects to a common sequence motif in thyroid hormone and estrogen response elements","volume":"54","author":"Glass","year":"1988","journal-title":"Cell"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1991","DOI":"10.1210\/me.2012.1227","article-title":"Estrogen receptors beta1 and beta2 have opposing roles in regulating proliferation and bone metastasis genes in the prostate cancer cell line PC3","volume":"26","author":"Dey","year":"2012","journal-title":"Mol. Endocrinol."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Langdon, S.P., Herrington, C.S., Hollis, R.L., and Gourley, C. (2020). Estrogen Signaling and Its Potential as a Target for Therapy in Ovarian Cancer. Cancers, 12.","DOI":"10.3390\/cancers12061647"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"3176","DOI":"10.1158\/0008-5472.CAN-17-3470","article-title":"Differential Effects of Estrogen Receptor beta Isoforms on Glioblastoma Progression","volume":"78","author":"Liu","year":"2018","journal-title":"Cancer Res."},{"key":"ref_46","first-page":"100060","article-title":"Polycystic ovary syndrome (PCOS) and genetic predisposition: A review article","volume":"3","author":"Ajmal","year":"2019","journal-title":"Eur. J. Obs. Gynecol. Reprod. Biol. X"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"881","DOI":"10.1210\/jc.2007-1650","article-title":"Granulosa cell survival and proliferation are altered in polycystic ovary syndrome","volume":"93","author":"Das","year":"2008","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"6182","DOI":"10.1210\/jc.2019-00780","article-title":"Gene Expression in Granulosa Cells from Small Antral Follicles From Women With or Without Polycystic Ovaries","volume":"104","author":"Owens","year":"2019","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"379","DOI":"10.3109\/09513590.2014.1001733","article-title":"Estrogen and progesterone receptor subtype expression in granulosa cells from women with polycystic ovary syndrome","volume":"31","author":"Artimani","year":"2015","journal-title":"Gynecol. Endocrinol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"3970","DOI":"10.1210\/jc.2017-00308","article-title":"Dysregulation of the Anti-Mullerian Hormone System by Steroids in Women with Polycystic Ovary Syndrome","volume":"102","author":"Pierre","year":"2017","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"5532","DOI":"10.1210\/jc.2002-020323","article-title":"Estrogen receptor alpha and beta expression in theca and granulosa cells from women with polycystic ovary syndrome","volume":"87","author":"Jakimiuk","year":"2002","journal-title":"J. Clin. Endocrinol. Metab."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/9\/5046\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:58:51Z","timestamp":1760162331000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/9\/5046"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,5,10]]},"references-count":51,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2021,5]]}},"alternative-id":["ijms22095046"],"URL":"https:\/\/doi.org\/10.3390\/ijms22095046","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,5,10]]}}}