Specific binding sites for atrial natriuretic factor (ANF) have been detected and localized in viable human spermatozoa through radioreceptor analysis and autoradiography, respectively. Radiotracer uptake was time and concentration dependent. Scatchard analysis of saturation data showed a single class of ANF receptors with a kd of 2.5 nM and a Bmax of 1.03 fmol/10(6) sperm 2.5 min-1, corresponding to about 620 molecules per sperm. Nonreducing SDS-PAGE analysis after covalent cross-linking of sperm bound 125I-ANF evidenced a single displaceable (i.e., specific) band with an apparent molecular weight of 135-140kD. In 125I-ANF bound spermatozoa, optical autoradiography showed an exclusive distribution of silver grains covering the midpiece region. The effects of ANF binding on ionic homeostasis and cyclic nucleotide metabolism, which modulate a number of sperm cellular processes, could make this factor play outstanding roles in gamete physiology.
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