Subcellular location of proteins with thiols groups, during epidydimal maturation.

CABRILLANA ME; MONCLUS MA; VINCENTI AE; FORNÉS MW; YUNES R

Mendoza

Congreso; XXVI Reunión Científica Anual de la Sociedad de Biología de Cuyo. I Reunión anual de la drección de Investigación, Ciencia y Técnica del Ministerio de Salud del Gobierno de Mendoza; 2008

Sociedad de Biología de Cuyo

During epididymal trip, the sperm proteins support a thiol oxidation. This redox changes from thiol to disulfides bonds in flagellar proteins are associating with the acquisition of sperm motility, capacitation, acrosome reaction and fertilization. Ours objectives were isolate different sperm subcellular fractions with proteins rich in this thiol groups. Using different detergent and sucrose gradient, we fractionated this cell such as head, acrosomal proteins, principal and terminal piece, outer dense fibers and fibrous sheath. Then we certificated the success of this procedure by electron microscopy. On the other hand we used monobromobimane, a fluorescent reactive that labels with protein`s thiols groups. We localized, by fluorescence microscopy, witch cellular fraction reacted. Others aliquots, were prepared for SDS-PAGE. About 27 kDa band, display a great fluorescence as well as the tail of the sperm. With this positive band we raised polyclonal antibody immunize rabbits. A positive reaction with sperms tail was observed. Finally, by immunological technique we are in condition to localize this protein/s in other tissues, and then to initiate the molecular characterization of this protein/s. These results allow us to find a probable function of these proteins in the sperm.