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Possible role of protein kinase C in regulation of flagellar motility and intracellular free Ca2+ concentration of fowl spermatozoa Protein kinase C and motility of fowl sperm 芦沢, 幸二 Ashizawa, Koji Katayama, S. Kobayashi, T. Tsuzuki, Y. 精子 sperm, spermatozoa, protein kinase C, calcium, motility, fowl The presence and possible role of protein kinase C in the regulation of fowl sperm functions were investigated. Immunoblot analysis of sperm extract using antibody to protein kinase C revealed a crossreacting protein of approximately 80 kDa. As the concentration of the protein kinase C activators N-（6-phenylhexyl）-5-chloro-1-naphthalenesulfonamide （SC-9） or 1-oleoyl-2-acetylglycerol （OAG） a protein kinase C activator, was increased, the motility of intact spermatozoa at 30°C was reduced. However, this inhibition of motility was reversed by reducing the concentrations of activators. Even in the presence of 1 mmol CaCl2 l-1, the addition of SC-9 and OAG inhibited the motility of intact spermatozoa. In contrast, the motility of demembranated spermatozoa was not inhibited by the addition of SC-9 or OAG at 30°C. On the other hand, the addition of 1-（5-isoquinolinesulfonyl）-2-methylpiperazine dihydrochloride （H-7）, a protein kinase C inhibitor, did not appreciably affect the motility of either intact or demembranated spermatozoa at 30°C. At 40°C, both intact and demembranated spermatozoa were almost immotile with or without any of the activators or inhibitor. Intracellular free Ca2+ concentrations, measured by means of a fluorescent Ca2+ indicator, fura-2, gradually increased after the addition of SC-9 and OAG, but no changes were observed in H-7-treated spermatozoa. These results suggest that endogenous protein kinase C is present in the cytoplasmic matrix and/or the membrane, but not retained in the axoneme, and that the activation of this enzyme may contribute to a decrease in the flagellar movement of fowl spermatozoa. Disclaimer. This is not the definitive version of record of this article. This manuscript has been accepted for publication in Reproduction, but the version presented here has not yet been copy edited, formatted or proofed. Consequently, neither Reproduction, the Society for Reproduction and Fertility nor its publisher, BioScientifica Ltd, accepts responsibility for any errors or omissions it may contain. The definitive version is now freely available at [insert DOI link]. 1994 Society for Reproduction and Fertility. 1994 eng journal article AM http://hdl.handle.net/10458/712 https://miyazaki-u.repo.nii.ac.jp/records/329 AA00705952 00224251 Journal of reproduction and fertility 101 3 511 517 https://miyazaki-u.repo.nii.ac.jp/record/329/files/ashizawa49.pdf application/pdf 1.5 MB 2020-06-21