Characterization and immunolocalization of beta-D-glucuronidase in mouse testicular germ cells and spermatozoa.
Spermatozoa released from the seminiferous tubules are terminally differentiated cells with no known synthetic activity. Their components are synthesized in the spermatogenic cells during spermatogenesis. In this study, we report the characterization and immunolocalization of beta-glucuronidase in mouse testicular germ cells and spermatozoa. The enzyme is an exoglycohydrolase with dual localization, being present in lysosomes and endoplasmic reticulum of several mouse and rat tissues. The purified germ cell preparations (spermatocytes, round spermatids, and condensed/elongated spermatids) when assayed for beta-glucuronidase activity showed that the spermatocytes contained five times more enzyme activity per cell than the spermatids. Polyacrylamide gel electrophoresis, carried out under native and denaturing conditions, demonstrated that the germ cells express only the lysosomal form of the enzyme (pI 5.5-6.0) with a subunit molecular mass of 74 kDa. Immunocytochemical studies revealed a positive reaction in the Golgi membranes, Golgi-associated vesicles, and lysosomes of late spermatocytes (pachytene spermatocytes) and a stage-specific localization during spermiogenesis. The forming or formed acrosome of the elongated spermatids (stages 9-16) and epididymal spermatozoa was highly immunopositive. Comparison of immunoprecipitation curves and kinetic properties of the enzyme present in spermatocytes and spermatozoa revealed no major differences. Taken together, our results demonstrate that beta-glucuronidase activities present in the lysosomes of spermatocytes and the sperm acrosome are kinetically and immunologically similar.
Laboratoire de Biologie Cellulaire, Université René Descartes, 45, rue des Saints-Pères, Paris Cedex 06, 75270, France.
A
A
Abou-Haïla
J P
JP
Fouquet
D R
DR
Tulsiani
D000818:Animals; D004789:Enzyme Activation; D004822:Epididymis / Q000737:chemistry / Q000201:enzymology / Q000648:ultrastructure; D019084:Fluorescent Antibody Technique, Indirect; D005966:Glucuronidase / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D013094:Spermatozoa / Q000737:chemistry / Q000201:enzymology* / Q000648:ultrastructure; D013737:Testis / Q000737:chemistry / Q000201:enzymology* / Q000648:ultrastructure
D005966:Glucuronidase