Abstract

Changes in the concentration of -SH groups on the human and rabbit spermatozoal membrane and during epididymal maturation were studied by means of a new fluorescent probe, carboxyphenylmaleimide (CPhM), which reacts specifically with -SH groups. Binding of CPhM did not modify oxygen uptake, motility, or viability of the sperm cells used, but produced a characteristic increase in fluorescence. By analysis of this increase it was possible to calculate the presence of 35 +/- 4.2 and 55 +/- 8 nmoles of exposed -SH groups/10(8) rabbit and human ejaculated spermatozoa, respectively. Caput epididymal cells bound significantly more CPhM than did cauda epididymal cells or ejaculated spermatozoa (155 +/- 22, 78 +/- 11, and 35 +/- 4.2 nmoles/10(8) cells, respectively, in rabbit cells; and 184, 110 +/- 18, and 55 +/- 8 nmoles/10(8) cells, respectively, in humans cells). In addition to the differences in number of exposed -SH groups observed between human and rabbit sperm cells, the behavior of these membrane-reactive groups when ethylenediaminetetraacetate and/or zinc were added to the incubation media indicates that the participation of membrane--SH groups in sperm physiology is species-specific.

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