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This study provides evidence that estrogen receptor alpha (ER\u03b1\/ESR1) phosphorylated at Ser309 is essential for efferent duct fluid reabsorption and epididymal luminal acidification to create and maintain the optimal luminal milieu in mice.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Abstract<\/jats:title>\n                    <jats:p>\n                      ER\u03b1 (ESR1) plays an important role in male reproduction and fertility. Its activity is modulated by phosphorylation of multiple amino acid residues. The ER\u03b1 phosphorylated at serine 305 (S305) in human cells (homologous with serine 309 in mice) induces ligand-independent ER\u03b1 activity. Here, we studied the effect of ER\u03b1 phosphorylation at S309 on the reproductive function of male mice. ER\u03b1309 (p.S309A,\n                      <jats:italic>ER\u03b1<\/jats:italic>\n                      <jats:sup>S309A<\/jats:sup>\n                      ) knock-in (KI) mice were generated by homologous recombination in mouse embryonic stem cells. The\n                      <jats:italic>ER\u03b1<\/jats:italic>\n                      <jats:sup>S309A<\/jats:sup>\n                      KI males were subfertile with decreased sperm motility, dilated lumen and thinned epithelium of efferent ducts. Ultrastructural observation showed that the area of endocytic vesicles and lysosomes in the non-ciliated cells and the number of cilia in the ciliated cells were significantly reduced in the efferent ducts of the\n                      <jats:italic>ER\u03b1<\/jats:italic>\n                      <jats:sup>S309A<\/jats:sup>\n                      KI mice. The pH value of the epididymal fluid was significantly higher in all regions of epididymis in the\n                      <jats:italic>ER\u03b1<\/jats:italic>\n                      <jats:sup>S309A<\/jats:sup>\n                      KI mice. Moreover, the expression of aquaporin 9 (AQP9) in the efferent ducts and carbonic anhydrase XII (CAR12) in the epididymis was decreased in\n                      <jats:italic>ER\u03b1<\/jats:italic>\n                      <jats:sup>S309A<\/jats:sup>\n                      KI mice. The results indicate that the ER\u03b1 phosphorylated at Ser309 participates in maintaining the unique luminal milieu by regulating the expression of critical regulators of fluid\/ion transport in the efferent ducts and epididymis.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1530\/rep-24-0109","type":"journal-article","created":{"date-parts":[[2025,1,7]],"date-time":"2025-01-07T07:04:47Z","timestamp":1736233487000},"update-policy":"https:\/\/doi.org\/10.1530\/crossmarkpolicy-11","source":"Crossref","is-referenced-by-count":3,"title":["ER\u03b1 phosphorylated at Ser309 participates in regulating luminal environment of efferent ducts and epididymis in mice"],"prefix":"10.1093","volume":"169","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0117-3208","authenticated-orcid":true,"given":"Jinhua","family":"Wei","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1051-2208","authenticated-orcid":true,"given":"Pang","family":"Cheng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3280-6565","authenticated-orcid":true,"given":"Binfang","family":"Ma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-7561-9446","authenticated-orcid":true,"given":"Xiao","family":"Feng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4735-4223","authenticated-orcid":true,"given":"Junxian","family":"Ma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-1795-0608","authenticated-orcid":true,"given":"Jie","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-8085-2302","authenticated-orcid":true,"given":"Yanqiu","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4944-4457","authenticated-orcid":true,"given":"Zhen","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","reference":[{"key":"bib1","series-title":"Mol Hum Reprod","article-title":"Motility of efferent duct cilia aids passage of sperm cells through the male reproductive system","volume":"27","author":"Aprea I","year":"2021","unstructured":"Aprea I, Nothe-Menchen T, Dougherty GW, et al. 2021 Motility of efferent duct cilia aids passage of sperm cells through the male reproductive system. 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