The removal of the sperm perinuclear theca and its association with the bovine oocyte surface during fertilization.
The perinuclear theca (PT) is a unique cytoskeletal structure whose anterior part is intercalated between the inner acrosomal membrane and the nuclear envelope of the mammalian sperm head and is important for spermiogenesis and stabilization of sperm structures (Oko and Maravei, Biol. Reprod. 50, 1000-1014, 1994; Oko and Maravei, Microsc. Res. Tech. 32, 520-532, 1995). Using immunofluorescence labeling of inseminated bovine oocytes and serial sectioning-ultrastructural analysis, we demonstrate that the PT is removed from the sperm nucleus following the loss of the sperm plasma membrane and the interaction of oocyte cortex with the PT. These events precede the development of the male pronucleus. The removal of the PT involves the elongated oocyte microvilli, rich in actin microfilaments, since it can be blocked by the microfilament-disrupting drug cytochalasin B. Reduction of disulfide bonds, which is a major factor supporting the disassembly of the sperm nucleus and accessory structures during mammalian fertilization, seems to exert little effect on the PT in vitro, as evidenced by the treatment of isolated bull sperm with the disulfide bond-reducing agent dithiothreitol. In vivo, intact bull sperm microinjected into mature oocytes do not undergo disassembly of the PT. Consequently, the decondensation of the sperm nucleus does not occur. These data suggest that the binding of the PT to the oocyte microvillar region and its removal from the sperm nucleus constitute an early step in mammalian fertilization, which is required for the conversion of the sperm nucleus into a male pronucleus.
Sutovsky
P
P
Department of Zoology, University of Wisconsin, 1117 West Johnson Street, Madison, Wisconsin 53706, USA.
Oko
R
R
Hewitson
L
L
Schatten
G
G
D000818:Animals; D000906:Antibodies / Q000276:immunology; D002417:Cattle; D003599:Cytoskeleton / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D004229:Dithiothreitol / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005306:Fertilization; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D008297:Male; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008871:Microvilli / Q000502:physiology / Q000648:ultrastructure; D009865:Oocytes / Q000502:physiology* / Q000648:ultrastructure; D013077:Sperm Head / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D013094:Spermatozoa / Q000502:physiology
D000906:Antibodies; D005456:Fluorescent Dyes; D004229:Dithiothreitol