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Spatial Distribution of Actin and Tubulin in Human Sperm Nuclear Matrix‐intermediate Filament Whole Mounts—A New Paradigm
Author(s) -
Kadam Kaushiki,
D'souza Serena,
Natraj Usha
Publication year - 2007
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20438
Subject(s) - cytoskeleton , sperm , microbiology and biotechnology , axoneme , biology , immunogold labelling , centriole , microtubule , treadmilling , actin , nuclear matrix , tubulin , ultrastructure , biophysics , anatomy , microfilament , chromatin , biochemistry , cell , flagellum , botany , dna , gene
Sperm is a highly differentiated cell streamlined for fertilization. The function is thus heavily dependent on the cytoskeletal organization. Conventional methods limit the appreciation and correlation of this intricate cytoskeletal filament network in the context of an entire sperm. Our recent successful localization of nonmuscle myosin IIA on sperm nuclear matrix‐intermediate filament (NM‐IF) preparations from fertile men by embedment‐free electron microscopy (EF‐EM), prompted us to investigate the antigenic distribution of two major cytoskeletal proteins—actin and tubulin. The NM‐IF preparations were subjected to a cocktail of buffered paraformaldehyde (2%) with a low concentration of glutaraldehyde (0.05%). These proteins were localized by indirect immunogold technique using EF‐EM on sperm NM‐IF whole mounts. Ultrastructure analysis revealed well preserved centrioles, outer dense fibers, axonemal filaments, and submitochondrial reticulum in the sperm NM‐IF. Immunoreactive actin was localized along the length of the sperm whereas β‐tubulin was present in the axoneme alone. The spatial distribution of actin and tubulin in normal human sperm NM‐IF reported here together with that of myosin on whole mount offers a powerful technique to understand sperm cytoskeletal supramolecular structure. Microsc. Res. Tech., 2007. © 2007 Wiley‐Liss, Inc.