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Acid glycohydrolases in rat spermatocytes, spermatids and spermatozoa: enzyme activities, biosynthesis and immunolocalization

Abstract

Mammalian sperm acrosomes contain several glycohydrolases that are thought to aid in the dispersion and digestion of vestments surrounding the egg. In this study, we have used multiple approaches to examine the origin of acrosome-associated glycohdyrdolases. Mixed spermatogenic cells, prepared from rat testis, were separated by unit gravity sedimentation. The purified germ cells (spermatocytes [SC], round spermatids [RS], and elongated/condensed spermatids [E/CS]) contained several glycohydrolase activities. Metabolic labeling in the cell culture, immunoprecipitation, and autoradiographic approaches revealed that β-D-galactosidase was synthesized in SC and RS in 88/90 kDa forms which undergo processing in a cell-specific manner. Immunohistochemical approaches demonstrated that the enzyme was localized in Golgi membranes/vesicles, and lysosome-like structures in SC and RS, and forming/formed acrosome of E/CS.

Abbreviations

MEM:

minimum essential medium

BSA:

bovine serum albumin

PBS:

phosphate buffered saline

SDS:

sodium dodecyl sulphate

PAGE:

polyacrylamide gel electrophoresis

PNP:

p-nitrophenyl

EKRB:

enriched Krebs-Ringer bicarbonate

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Correspondence to Daulat R. P. Tulsiani.

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Published December 3, 2001

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Abou-Haila, A., Tulsiani, D.R.P. Acid glycohydrolases in rat spermatocytes, spermatids and spermatozoa: enzyme activities, biosynthesis and immunolocalization. Biol Proced Online 3, 35–42 (2001). https://doi.org/10.1251/bpo21

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Indexing terms

  • Sperm acrosome
  • male germ cells
  • β-D-galactosidase
  • acrosmal glycohydrolases
  • mammalian fertilization