{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T15:53:49Z","timestamp":1759938829445},"reference-count":39,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2013,1,2]],"date-time":"2013-01-02T00:00:00Z","timestamp":1357084800000},"content-version":"vor","delay-in-days":909,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Journal of Andrology"],"published-print":{"date-parts":[[2010,7,8]]},"abstract":"<jats:p><jats:bold>ABSTRACT: <\/jats:bold> This study investigates the dynamics of serine\/threonine (S\/T) protein phosphorylation in sperm incubated under capacitating (C) conditions using the boar as a model system. For the first time, this approach has identified multiple dephosphorylation events that occur in a bicarbonate\u2010dependent fashion. Different phospho\u2010(S\/T) kinase substrate antibodies were used, and dephosphorylation of 5 S\/T phosphoproteins was observed in C sperm compared with noncapacitated (N) cells. Specifically, dephosphorylation of 96\u2010, 90\u2010, 64\u2010, and 55\u2010kd proteins was detected by immunoblotting using 2 phospho\u2010Akt substrate antibodies and a phosphoprotein kinase A substrate antibody. In addition, dephosphorylation of a 105\u2010kd protein was detected using a phospho\u2010ATM\/ATR substrate antibody. In contrast, no dephosphorylation was observed using a phosphoprotein kinase C substrate antibody, and increased tyrosine phosphorylation of 32\u2010 and 20\u2010kd proteins was detected in C compared with N sperm. Immunolocalization experiments revealed subtle changes in the pattern expression as well as a reduction of phosphorylation in C sperm. Whereas sperm incubated in N medium containing dibutyryl cAMP (dbcAMP) and 3\u2010isobutyl\u20101\u2010methylxanthine (IBMX) did not show protein dephosphorylation, incubation in C medium with dbcAMP\/IBMX showed dephosphorylation as well as increased phosphorylation of other proteins (p68, p51, and p29). Finally, calyculin A, a phosphatase inhibitor, prevented dephosphorylation of p96, p90, p64, and p55 but not p105. Based on these data, we propose 2 pathways of protein dephosphorylation that are active during capacitation and independent of cAMP. Together, this provides direct evidence for more complex S\/T phosphorylation dynamics than has been previously described for sperm undergoing capacitation.<\/jats:p>","DOI":"10.2164\/jandrol.109.008383","type":"journal-article","created":{"date-parts":[[2010,9,8]],"date-time":"2010-09-08T18:27:08Z","timestamp":1283970428000},"page":"393-405","source":"Crossref","is-referenced-by-count":7,"title":["Bicarbonate\u2010Dependent Serine\/Threonine Protein Dephosphorylation in Capacitating Boar Spermatozoa"],"prefix":"10.1002","volume":"31","author":[{"given":"Fahima A.","family":"Alnagar","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paul","family":"Brennan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ian 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