{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,18]],"date-time":"2026-04-18T15:15:34Z","timestamp":1776525334044,"version":"3.51.2"},"reference-count":40,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2004,7,29]],"date-time":"2004-07-29T00:00:00Z","timestamp":1091059200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecular Reproduction Devel"],"published-print":{"date-parts":[[2004,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>A cAMP\u2010induced increase of tyrosine\u2010phosphorylated proteins is involved in the expression of fertilizing ability in mammalian spermatozoa. We (Harayama, 2003: J Androl 24:831\u2013842) reported that incubation of boar spermatozoa with a cell\u2010permeable cAMP analog (cBiMPS) increased a 32\u2010kDa tyrosine\u2010phosphorylated protein (TyrP32). The purpose of this study is to characterize the signaling cascades that regulate the cAMP\u2010induced increase of TyrP32. We examined effects of tyrosine kinase inhibitor (lavendustin A), tyrosine phosphatase inhibitor (Na<jats:sub>3<\/jats:sub>VO<jats:sub>4<\/jats:sub>), cell\u2010permeable calcium chelator (BAPTA\u2010AM), and cholesterol acceptor (methyl\u2010\u03b2\u2010cyclodextrin: MBC) on the increase of TyrP32 and the change and loss of acrosomes in boar spermatozoa. The spermatozoa were used for detection of tyrosine\u2010phosphorylated proteins by Western blotting and indirect immunofluorescence and for examination of acrosomal integrity by Giemsa staining. At least eight tyrosine\u2010phosphorylated proteins including TyrP32 exhibited the cAMP\u2010dependent increase during incubation with cBiMPS. In many proteins of them, this increase was reduced by lavendustin A but was enhanced by Na<jats:sub>3<\/jats:sub>VO<jats:sub>4<\/jats:sub>. In contrast, the cAMP\u2010induced increase of TyrP32 was abolished by Na<jats:sub>3<\/jats:sub>VO<jats:sub>4<\/jats:sub> but was hardly affected by lavendustin A. Giemsa staining showed that the increase of spermatozoa with weakly Giemsa\u2010stained acrosomes (severely damaged acrosomes) or without acrosomes was correlative to the cAMP\u2010induced increase of TyrP32. Moreover, the lack of calcium chloride in the incubation medium or pretreatment of spermatozoa with BAPTA\u2010AM blocked the change and loss of acrosomes and the increase of TyrP32, suggesting these events are dependent on the extracellular and intracellular calcium. On the other hand, incubation of spermatozoa with MBC in the absence of cBiMPS could mimic the change and loss of acrosomes and increase of TyrP32 without increase of other tyrosine\u2010phosphorylated proteins. Based on these results, we conclude that the cAMP\u2010induced increase of TyrP32 is regulated by a unique mechanism that may be linked to the calcium\u2010dependent change and loss of acrosomes. Mol. Reprod. Dev. 69: 194\u2013204, 2004. \u00a9 2004 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/mrd.20099","type":"journal-article","created":{"date-parts":[[2004,7,29]],"date-time":"2004-07-29T21:25:29Z","timestamp":1091136329000},"page":"194-204","source":"Crossref","is-referenced-by-count":38,"title":["A unique mechanism for cyclic adenosine 3\u2032,5\u2032\u2010monophosphate\u2010induced increase of 32\u2010kDa tyrosine\u2010phosphorylated protein in boar 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