SOLID-PHASE CELLULAR LECTIN ASSAY TO STUDY SURFACE LECTIN-BINDING SITES ON RHESUS-MONKEY SPERMATOZOA

Here we report the binding of lectins (Con A, PNA and WGA) to cauda ep ididymal and ejaculated rhesus monkey spermatozoa using a solid-phase cellular lectin enzyme immunoassay (CLEIA). For standardization of the assay, optimum dilutions of lectins, enzyme conjugated antibodies, in hibitor saccharides and sperm number for coating to microtitre plate w ells were established. Using radioiodinated lectins, radio-lectin assa y was also established but correlation analysis showed CLEIA had highe r sensitivity compared to radio-lectin assay (CRLA). Using CLEIA, WGA binding to ejaculated spermatozoa showed a significant increase compar ed to cauda epididymal spermatozoa. Using FITC labelled lectins, Con A , PNA and WGA were localized mainly in the acrosomal region of caudal and ejaculated spermatozoa while the post-acrosomal region did not sho w any fluorescence. The solid phase lectin immunoassay developed in th is study with a relatively small number of sperm is simple to perform, sensitive and specific and can be used to screen antisera and to anal yse changes in lectin binding to cell surfaces.