{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T16:28:23Z","timestamp":1791822503879,"version":"4.3.4"},"reference-count":41,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T00:00:00Z","timestamp":1783555200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003500","name":"University of Padua","doi-asserted-by":"publisher","award":["ANDR_BIRD24_01"],"award-info":[{"award-number":["ANDR_BIRD24_01"]}],"id":[{"id":"10.13039\/501100003500","id-type":"DOI","asserted-by":"publisher"}]},{"award":["ANDR_BIRD24_01"],"award-info":[{"award-number":["ANDR_BIRD24_01"]}],"id":[{"id":"https:\/\/ror.org\/00240q980","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>The anion exchanger 1 (AE1), traditionally known for its role in erythrocyte anion transport and acid\u2013base homeostasis, has recently been identified in non-erythroid tissues, suggesting broader physiological functions. In the present study, we investigated for the first time the expression and potential role of AE1 in human ovarian granulosa cells (GCs) obtained from women with endometriosis (ENDO-GCs) or male factor infertility controls (MF-GCs). Cells were cultured in the presence of follicular fluid derived from control (FF-MF) or endometriosis patients (FF-ENDO), and DIDS-sensitive anion exchange activity was pharmacologically inhibited using 4,4\u2032-diisothiocyanatostilbene-2,2\u2032-disulfonic acid (DIDS). AE1 expression was evaluated at both protein and transcript levels together with markers of proliferation, inflammation, and steroidogenesis. The results demonstrate that AE1 is constitutively expressed in GCs and may contribute to granulosa cell homeostasis. Inhibition of DIDS-sensitive anion exchange activity inhibited cell proliferation, shifted cell morphology toward a fibroblast-like phenotype, and reduced estradiol and progesterone secretion and inflammatory (IL-6) gene transcription. Notably, MF-GCs cultured in FF-MF exhibited compensatory upregulation of AE1, whereas ENDO-GCs and cells exposed to FF-ENDO showed impaired adaptive responses and generalized transcriptional suppression. These findings provide preliminary evidence supporting a role for DIDS-sensitive anion exchange activity in granulosa cell physiology and warrant further studies to clarify the specific contribution of AE1 to follicular dysfunction associated with endometriosis. However, the specific mechanistic contribution of AE1 could not be established and will require further functional and genetic investigations.<\/jats:p>","DOI":"10.3390\/biom16071004","type":"journal-article","created":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T11:55:32Z","timestamp":1783598132000},"page":"1004","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Expression of Anion Exchanger 1 (AE1) and Its Potential Involvement in Human Granulosa Cell Physiology: An In Vitro Pilot Study"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4771-031X","authenticated-orcid":false,"given":"Loris","family":"Marin","sequence":"first","affiliation":[{"name":"Department of Women\u2019s and Children\u2019s Health, University of Padova, 35128 Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chiara","family":"Sabbadin","sequence":"additional","affiliation":[{"name":"Endocrine Unit, Department of Medicine DIMED, University of Padova, 35128 Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5548-9785","authenticated-orcid":false,"given":"Claudia Maria","family":"Radu","sequence":"additional","affiliation":[{"name":"Thrombotic and Haemorrhagic Diseases Unit, Department of Medicine-DIMED, University of Padova, 35128 Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3279-9000","authenticated-orcid":false,"given":"Paola","family":"Brun","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine DMM, University of Padova, 35128 Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carolina","family":"Frison","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine DMM, University of Padova, 35128 Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4269-4269","authenticated-orcid":false,"given":"Giuseppe","family":"Gullo","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Villa Sofia Cervello Hospital, University of Palermo, 90146 Palermo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6433-5155","authenticated-orcid":false,"given":"Decio","family":"Armanini","sequence":"additional","affiliation":[{"name":"Endocrine Unit, Department of Medicine DIMED, University of Padova, 35128 Padova, Italy"},{"name":"Studium Patavinum, University of Padova, 35122 Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4238-3153","authenticated-orcid":false,"given":"Luciana","family":"Bordin","sequence":"additional","affiliation":[{"name":"Department of Women\u2019s and Children\u2019s Health, University of Padova, 35128 Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0390-6823","authenticated-orcid":false,"given":"Eugenio","family":"Ragazzi","sequence":"additional","affiliation":[{"name":"Studium Patavinum, University of Padova, 35122 Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guido","family":"Ambrosini","sequence":"additional","affiliation":[{"name":"Department of Women\u2019s and Children\u2019s Health, University of Padova, 35128 Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9428-4363","authenticated-orcid":false,"given":"Alessandra","family":"Andrisani","sequence":"additional","affiliation":[{"name":"Department of Women\u2019s and Children\u2019s Health, University of Padova, 35128 Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"963","DOI":"10.1172\/JCI41350","article-title":"The Ovary: Basic Biology and Clinical Implications","volume":"120","author":"Richards","year":"2010","journal-title":"J. Clin. Investig."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"489","DOI":"10.1530\/REP-10-0094","article-title":"Theca: The Forgotten Cell of the Ovarian Follicle","volume":"140","author":"Young","year":"2010","journal-title":"Reproduction"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1530\/rep.1.01074","article-title":"TGF-\u03b2 Superfamily Members and Ovarian Follicle Development","volume":"132","author":"Knight","year":"2006","journal-title":"Reproduction"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Marin, L., Sabbadin, C., Faggin, G., Radu, C.M., Armanini, D., Paccagnella, M., Salata, C., Bordin, L., Ragazzi, E., and Ambrosini, G. (2025). Endometriotic Follicular Fluid Affects Granulosa Cells\u2019 Morphology and Increases Duplication Rate and Connexin-43 Expression. Biomolecules, 15.","DOI":"10.3390\/biom15040561"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Marin, L., Sabbadin, C., Radu, C.M., Brun, P., Frison, C., Gullo, G., Armanini, D., Bordin, L., Ragazzi, E., and Ambrosini, G. (2025). Progesterone and IL-6 Expression Are Modulated by Follicular Fluid in Granulosa Cell Cultures. Biomolecules, 15.","DOI":"10.3390\/biom15121646"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1186\/1477-7827-7-40","article-title":"Follicular Fluid Content and Oocyte Quality: From Single Biochemical Markers to Metabolomics","volume":"7","author":"Revelli","year":"2009","journal-title":"Reprod. Biol. Endocrinol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1186\/s13048-024-01551-9","article-title":"Unraveling the Complexity of Follicular Fluid: Insights into Its Composition, Function, and Clinical Implications","volume":"17","author":"Pan","year":"2024","journal-title":"J. Ovarian Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1021","DOI":"10.1095\/biolreprod.109.082941","article-title":"Formation of the Ovarian Follicular Antrum and Follicular Fluid1","volume":"82","author":"Rodgers","year":"2010","journal-title":"Biol. Reprod."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"735","DOI":"10.1007\/s10815-018-1143-3","article-title":"Influence of Follicular Fluid and Cumulus Cells on Oocyte Quality: Clinical Implications","volume":"35","author":"Giorgi","year":"2018","journal-title":"J. Assist. Reprod. Genet."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e9","DOI":"10.1017\/erm.2016.4","article-title":"Protein Pathways Working in Human Follicular Fluid: The Future for Tailored IVF?","volume":"18","author":"Bianchi","year":"2016","journal-title":"Expert Rev. Mol. Med."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Gebremedhn, S., Ali, A., Gad, A., Prochazka, R., and Tesfaye, D. (2020). Extracellular Vesicles as Mediators of Environmental and Metabolic Stress Coping Mechanisms During Mammalian Follicular Development. Front. Vet. Sci., 7.","DOI":"10.3389\/fvets.2020.602043"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1023","DOI":"10.1016\/j.rbmo.2020.09.002","article-title":"The Follicular Fluid Metabolome Differs According to the Endometriosis Phenotype","volume":"41","author":"Santulli","year":"2020","journal-title":"Reprod. Biomed. Online"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1076","DOI":"10.1093\/humrep\/16.6.1076","article-title":"Follicular Fluid as a Favourable Environment for Endometrial and Endometriotic Cell Growth in Vitro","volume":"16","author":"Somigliana","year":"2001","journal-title":"Hum. Reprod."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Riemma, G., Lagan\u00e0, A.S., Schiattarella, A., Garzon, S., Cobellis, L., Autiero, R., Licciardi, F., Della Corte, L., La Verde, M., and De Franciscis, P. (2020). Ion Channels in The Pathogenesis of Endometriosis: A Cutting-Edge Point of View. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21031114"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1507","DOI":"10.1016\/j.bbamem.2016.03.030","article-title":"Band 3, the Human Red Cell Chloride\/Bicarbonate Anion Exchanger (AE1, SLC4A1), in a Structural Context","volume":"1858","author":"Reithmeier","year":"2016","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/j.mam.2012.10.008","article-title":"The SLC4 Family of Bicarbonate Transporters","volume":"34","author":"Romero","year":"2013","journal-title":"Mol. Asp. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1002\/pst.185","article-title":"Sample Size of 12 per Group Rule of Thumb for a Pilot Study","volume":"4","author":"Julious","year":"2005","journal-title":"Pharm. Stat."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"The ESHRE Working Group on Ultrasound in ART, D\u2019Angelo, A., Panayotidis, C., Amso, N., Marci, R., Matorras, R., Onofriescu, M., Turp, A.B., Vandekerckhove, F., and Veleva, Z. (2019). Recommendations for Good Practice in Ultrasound: Oocyte Pick Up. Hum. Reprod. Open, 2019, hoz025.","DOI":"10.1093\/hropen\/hoz025"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Kalli, A.C., and Reithmeier, R.A.F. (2022). Organization and Dynamics of the Red Blood Cell Band 3 Anion Exchanger SLC4A1: Insights From Molecular Dynamics Simulations. Front. Physiol., 13.","DOI":"10.3389\/fphys.2022.817945"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Don\u00e0, G., Tibaldi, E., Andrisani, A., Ambrosini, G., Sabbadin, C., Pagano, M.A., Brunati, A.M., Armanini, D., Ragazzi, E., and Bordin, L. (2020). Human Sperm Capacitation Involves the Regulation of the Tyr-Phosphorylation Level of the Anion Exchanger 1 (AE1). Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21114063"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1007\/s00424-003-1180-2","article-title":"The SLC4 Family of HCO3 ? Transporters","volume":"447","author":"Romero","year":"2004","journal-title":"Pfl\u00fcgers Arch. Eur. J. Physiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"11917","DOI":"10.1021\/bi048622a","article-title":"Relocation of the Disulfonic Stilbene Sites of AE1 (Band 3) on the Basis of Fluorescence Energy Transfer Measurements","volume":"43","author":"Knauf","year":"2004","journal-title":"Biochemistry"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"7888","DOI":"10.1021\/bi00372a015","article-title":"Molecular Mechanisms of Band 3 Inhibitors. 1. Transport Site Inhibitors","volume":"25","author":"Falke","year":"1986","journal-title":"Biochemistry"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"418","DOI":"10.4103\/1008-682X.181816","article-title":"Chloride Channels Are Involved in Sperm Motility and Are Downregulated in Spermatozoa from Patients with Asthenozoospermia","volume":"19","author":"Liu","year":"2017","journal-title":"Asian J. Androl."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1007\/s13577-020-00439-2","article-title":"Characterization of Stem Cells from Human Ovarian Follicular Fluid; a Potential Source of Autologous Stem Cell for Cell-Based Therapy","volume":"34","author":"Rungsiwiwut","year":"2021","journal-title":"Hum. Cell"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Jozkowiak, M., Hutchings, G., Jankowski, M., Kulcenty, K., Mozdziak, P., Kempisty, B., Spaczynski, R.Z., and Piotrowska-Kempisty, H. (2020). The Stemness of Human Ovarian Granulosa Cells and the Role of Resveratrol in the Differentiation of MSCs\u2014A Review Based on Cellular and Molecular Knowledge. Cells, 9.","DOI":"10.3390\/cells9061418"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1097\/00062752-200203000-00009","article-title":"Band 3 Anion Exchanger and Its Involvement in Erythrocyte and Kidney Disorders","volume":"9","author":"Tanner","year":"2002","journal-title":"Curr. Opin. Hematol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e13796","DOI":"10.1111\/apha.13796","article-title":"Role of SLC4 and SLC26 Solute Carriers during Oxidative Stress","volume":"235","author":"Remigante","year":"2022","journal-title":"Acta Physiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"928","DOI":"10.1080\/09513590.2017.1332177","article-title":"Dapsone Hydroxylamine-Mediated Alterations in Human Red Blood Cells from Endometriotic Patients","volume":"33","author":"Andrisani","year":"2017","journal-title":"Gynecol. Endocrinol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1016\/j.rbmo.2014.01.016","article-title":"Increased Oxidation-Related Glutathionylation and Carbonic Anhydrase Activity in Endometriosis","volume":"28","author":"Andrisani","year":"2014","journal-title":"Reprod. Biomed. Online"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1616","DOI":"10.1016\/j.fertnstert.2009.10.038","article-title":"Evaluation of Erythrocyte Band 3 Phosphotyrosine Level, Glutathione Content, CA-125, and Human Epididymal Secretory Protein E4 as Combined Parameters in Endometriosis","volume":"94","author":"Bordin","year":"2010","journal-title":"Fertil. Steril."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"7265238","DOI":"10.1155\/2017\/7265238","article-title":"Oxidative Stress and Endometriosis: A Systematic Review of the Literature","volume":"2017","author":"Scutiero","year":"2017","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"7924021","DOI":"10.1155\/2018\/7924021","article-title":"The Role of Oxidative Stress and Membrane Transport Systems during Endometriosis: A Fresh Look at a Busy Corner","volume":"2018","author":"Vitale","year":"2018","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1016\/j.reprotox.2013.08.005","article-title":"Markers of Oxidative Stress in Follicular Fluid of Women with Endometriosis and Tubal Infertility Undergoing IVF","volume":"42","author":"Singh","year":"2013","journal-title":"Reprod. Toxicol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1656","DOI":"10.1177\/1933719116674077","article-title":"Oxidative Stress in Granulosa-Lutein Cells From In Vitro Fertilization Patients","volume":"23","author":"Rotoli","year":"2016","journal-title":"Reprod. Sci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1002\/mrd.22943","article-title":"Oocyte Oxidative DNA Damage May Be Involved in Minimal\/Mild Endometriosis-related Infertility","volume":"85","author":"Ferriani","year":"2018","journal-title":"Mol. Reprod. Dev."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"619","DOI":"10.1530\/REP-09-0112","article-title":"Regulation of Intracellular pH during Oocyte Growth and Maturation in Mammals","volume":"138","author":"FitzHarris","year":"2009","journal-title":"Reproduction"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1677\/joe.0.1640247","article-title":"The Role of the StAR Protein in Steroidogenesis: Challenges for the Future","volume":"164","author":"Stocco","year":"2000","journal-title":"J. Endocrinol."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Midzak, A., and Papadopoulos, V. (2016). Adrenal Mitochondria and Steroidogenesis: From Individual Proteins to Functional Protein Assemblies. Front. Endocrinol., 7.","DOI":"10.3389\/fendo.2016.00106"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"4575","DOI":"10.1210\/en.2003-0268","article-title":"Intracellular Transport of Calcium from Plasma Membrane to Mitochondria in Adrenal H295R Cells: Implication for Steroidogenesis","volume":"144","author":"Resin","year":"2003","journal-title":"Endocrinology"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1016\/0022-4731(89)90221-5","article-title":"Sodium-Dependent Regulation of Steroidogenesis in Rat Granulosa Cells: Possible Involvement of a Na+\/H+ Antiport","volume":"32","author":"Li","year":"1989","journal-title":"J. Steroid Biochem."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/16\/7\/1004\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T12:24:08Z","timestamp":1783599848000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/16\/7\/1004"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,9]]},"references-count":41,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["biom16071004"],"URL":"https:\/\/doi.org\/10.3390\/biom16071004","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,9]]}}}