Cytoskeletal differences between stereocilia of the human sperm passageway and microvilli/stereocilia in other locations

Abstract

Background: Stereocilia of the human ductus epididymidis and ductus deferens display unique features in that they arise from an apical cell protrusion (hillock) and contain thick stem portions which are interconnected by cytoplasmic bridges. The molecular basis for this unique fusion and branching pattern is hitherto unknown. These morphologic specialties led us to study the cytoskeleton of male spermway stereocilia with respect to the major proteins that constitute the supportive cytoskeleton of intestinal microvilli and inner ear stereocilia.

Methods: Samples of the human epididymidis and ductus deferens were studied by immunoblotting and immunocytochemistry at the light and electron microscope levels.

Results: Spermway stereocilia are supported by an internal actin filament bundle crosslinked by fimbrin and associated with the membrane linker molecule ezrin. The stem portions and hillock area are supplied with the crossbridge forming molecule alpha-actinin. Spermway stereocilia differ from brush border microvilli of the intestine, kidney, and ductuli efferentes by the lack of the second bundling protein villin and the unusual expression of alpha-actinin in the stem region. They resemble inner ear stereocilia by the presence of fimbrin and absence of villin, but differ from them by expression of ezrin and alpha-actinin. Thus, the main molecular difference between spermway stereocilia and stereocilia/microvilli of other locations is the presence of alpha-actinin in their stem portion and the hillock area.

Conclusions: Since alpha-actinin can form crossbridges between adjacent actin filaments (bundles) at longer distances than the other crosslinker of the stereocilium core bundle, fimbrin, we assume that alpha-actinin is essential for both the formation of the stem portions of spermway stereocilia and for the generation of their striking branching pattern. A developmentally regulated temporal sequence of expression of fimbrin and alpha-actinin might control the unique architecture of spermway stereocilia.