Ultrastructural localization of acrosome reaction‐inducing substance (ARIS) on sperm of the starfish Asterias amurensis

Using colloidal gold tagged ligands we have identified the ultrastructural site of ARIS binding to intact and acrosome‐reacted starfish sperm. In intact sperm, colloidal gold conjugated ARIS was specifically localized to a single domain (0.1–0.3μ in diameter) on the plasma membrane. This site was located on the anterior‐lateral aspect of the sperm head, that is, just peripheral to the region occupied by the acrosomal vesicle and periacrosomal components. When sperm were labeled with colloidal gold conjugated ARIS, washed to remove unbound label, and then induced to undergo the acrosome reaction, the labeled patch remained associated with the plasma membrane and was positioned just lateral to the acrosomal process. However, when sperm were suspended in labeled ARIS and induced to undergo the acrosome reaction, label was observed along the entire anterior aspect of the sperm head with the exception of the acrosomal process. Labeling along the entire anterior aspect of the sperm head in this case was deemed to be nonspecific and due to binding of colloidal gold tagged molecules to components formerly located within the acrosomal vesicle, as the same pattern was obtained using colloidal gold tagged bovine serum albumin. Quantitative and qualitative aspects of ARIS binding observed here by electron microscopy are in agreement with measured binding characteristics previously reported (Ushiyama et al., 1993a: Zygote 1:121–127; Ushiyama et al., 1993b: J Reprod Dev 39:53–54), and indicate that the site of labeled ARIS binding represents a specific plasma membrane domain occupied by ARIS receptors. © 1995 Wiley‐Liss, Inc.

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