Cationic probes: Specificity of distribution of metal-binding sites in bovine sperm

Salts of transition elements that alter the rate of sperm cell movement act at or near calcium-binding sites. After living bull sperm cells had been preincubated in VO43−, Ni2+, Zn2+, Mn2+, and also La3+, they were then fixed. Crisply defined organelles and the absence of particulate deposits in the morphological controls contrasted sharply with the treated specimens; the latter contained regions of increased electron density, the nature and distribution of which depended on the test substance, reflecting the differential affinities of the specific ions. La3+ formed fine dense areas, mainly at the exocytic surface of the plasma membrane. VO43− marks the cell surface but also left particulate densities within the cell. Ni2+ caused a nearly uniformly dense deposit at the surface and on the satellite fibers and axonemal microtubules. Zn2+ formed less uniform but coarser deposits, while in Mn2+ the distribution was similar to that in Zn2+ but much denser in the axonemal matrix and on the satellite fibers. Verapamil restricted the size and number of the opacities, while procaine permitted a similar distribution of slightly larger size reaction product. The differences in size and distribution of the enhanced densities were consistent and replicable for the individual assay substances. Vanadate, which specifically inhibits Na, K-ATPase, bound to ouabain-sensitive enzyme loci, however, completely disrupting the axonemal complex. This suggests that an important role of dynein in flagellar motion may relate to intracellular transport of Ca2+.