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F‐Actin in acrosome‐reacted boar spermatozoa
Author(s) -
CastellaniCeresa L.,
Brivio M. F.,
Radaelli G.
Publication year - 1992
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.1080330114
Subject(s) - acrosome , acrosome reaction , boar , biology , biophysics , microbiology and biotechnology , immunoelectron microscopy , phalloidin , actin , sperm , anatomy , cytoskeleton , biochemistry , semen , cell , immunology , antibody , botany
Biochemical and immunoelectron microscopic methods have been used to analyze the distribution of actin in boar spermatozoa and its state of aggregation before and after acrosome reaction. F‐actin was detected on sperm head and tail by electron microscopy using an improved phalloidin probe: incubation with a fluorescein–phalloidin complex and an anti‐fluorescein antibody, followed by labeling with protein A‐gold complex. Gold particles, indicating the presence of F‐actin, were localized on the sperm surface of the acrosome‐reacted spermatozoa. Specific labeling was localized (1) between the outer acrosomal membrane and the plasma membrane in the equatorial region, (2) between the outer surface of the fibrous sheath and the plasma membrane in the postacrosomal region, (3) around the connecting piece and the neck region, and (4) on the external surface of the fibrous sheath in the principal piece of the tail. Furthermore, after NP‐40 extraction, the SDS‐PAGE revealed a difference in solubility between reacted and unreacted boar spermatozoa, reflecting actin polymerization. We conclude that most actin in the acrosome reacted boar spermatozoa is polymeric. © 1992 Wiley‐Liss, Inc.