Localization of sulfatoxygalactosylacylalkylglycerol at the surface of rat testicular germinal cells by immunocytochemical techniques: pH dependence of a nonimmunological reaction between immunoglobulin and germinal cells.

Abstract

The synthesis of sulfatoxygalactosylacylalkylglycerol (SGG) is a marker of germinal cell differentiation during spermatogenesis. Antibodies raised against this lipid have been used to visualize SGG on the surfaces of rat spermatocytes and spermatids. An ionic interaction between SGG and immunoglobulin was shown to occur at physiological pH, resulting in high fluorescence backgrounds for control cells treated with nonimmune sera. Immunofluorescence was therefore performed at alkaline pH such that this interaction was much reduced or eliminated. A method was also developed to detect surface-bound complement fixed in the presence of anti-SGG. SGG was found to be mobile within the plane of the membrane, undergoing ligand-induced "patching" and occasional "capping." However, this phenomenon was independent of temperature.