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Calcium-Dependent Actin Filament-Severing Protein Scinderin Levels and Localization in Bovine Testis, Epididymis, and Spermatozoa¹

^a Department of Pathology and Cell Biology, Faculty of Medicine, Université de Montréal, P. Québec, Canada H3T 1J4 ^b Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, Ontario, Canada K1H 8M5

We assessed the levels and localization of the actin filament-severing protein scinderin, in fetal and adult bovine testes, and in spermatozoa during and following the epididymal transit. We performed immunoblots on seminiferous tubules and interstitial cells isolated by enzymatic digestion, and on bovine chromaffin cells, spermatozoa, aorta, and vena cava. Immunoperoxidase labeling was done on Bouin's perfusion-fixed testes and epididymis tissue sections, and on spermatozoa. In addition, immunofluorescence labeling was done on spermatozoa. Immunoblots showed one 80-kDa band in chromaffin cells, fetal and adult tubules, interstitial cells, spermatozoa, aorta, and vena cava. Scinderin levels were higher in fetal than in adult seminiferous tubules but showed no difference between fetal and adult interstitial cells. Scinderin levels were higher in epididymal than in ejaculated spermatozoa. Scinderin was detected in a region corresponding with the subacrosomal space in the round spermatids and with the acrosome in the elongated spermatids. In epididymal spermatozoa, scinderin was localized to the anterior acrosome and the equatorial segment, but in ejaculated spermatozoa, the protein appeared in the acrosome and the post-equatorial segment of the head. In Sertoli cells, scinderin was detected near the cell surface and within the cytoplasm, where it accumulated near the base in a stage-specific manner. In the epididymis, scinderin was localized next to the surface of the cells; in the tail, it collected near the base of the principal cells. In Sertoli cells and epididymal cells, scinderin may contribute to the regulation of tight junctional permeability and to the release of the elongated spermatids by controlling the state of perijunctional actin. In germ cells, scinderin may assist in the shaping of the developing acrosome and influence the fertility of the spermatozoa.

¹ This work was supported in part by Population Council Award B97.071P–9/ICMC to R.M.P.; CORPAQ grant to R.M.P. and M.L.V.; MRC Grant MT-12879 to M.L.V.; MRC grant MT-11169 to J.M.T. M.L.V. is also funded by a scholarship from Fonds de la Recherche en santé du Québec.

² Correspondence: R.-Marc Pelletier, Université de Montréal, Faculty of Medicine, Department of Pathology and Cell Biology, Pavillon Principal, 2900 Edouard-Montpetit blvd. Montréal, P. Québec, Canada H3T 1J4. FAX: 514 485 7932; pellemar{at}ere.umontreal.ca

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