RESUMO
Understanding the signal transduction processes that occur during oocyte maturation and fertilization requires knowledge of the constituent proteins from the cell surface to relevant intracellular compartments. To identify starfish oocyte and egg cell surface proteins, a biotinylation method was adapted from prior protocols using B cells, leukocytes, mouse oocytes, and sea urchin eggs (Cole et al. Mol Immunol 24:699-705, 1987; Flaherty and Swann NJ. Mol Reprod Dev 35:285-292, 1993; Haley and Wessel. Dev Biol 272:191-202, 2004; Hurley et al. J Immunol Methods 85:195-202, 1985). This method utilizes the water-soluble Sulfo-NHS-Biotin, which does not cross the egg plasma membrane. The process of biotinylation does not appear to have any effect on the process of oocyte maturation or fertilization. Furthermore, it can be used with either vitelline-intact or vitelline-free oocytes and allows the proteins to be visualized successfully through immunoblotting, immunoprecipitation, or by scanning confocal microscopy.
