Expression of ebaf in Pregnant Rat Oviduct. Sinthesys and Tissue Distribution

MARTIN E. ARGAÑARAZ; PABLO A. VALDECANTOS; DORA C. MICELI

Rosario, Santa Fé

Congreso; XLII Reunión anual de la Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular - SAIB; 2006

Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular - SAIB

Ebaf, a member of the TGF-b family, is involved in the regulation of other citokines of this family such us nodal, activin, BMPs, TGF-b1 and Vg1. Previously we had reported the identification, isolation, cloning and characterization of the temporal expression levels of ebaf in the oviduct during the early pregnancy of the rat. The aims of the present work were to confirm the synthesis of Ebaf protein in the oviduct and to study its tissue localization. The analyses of the deducted aminoacidic sequence (Accession No. AAV31601.1) revealed a proprotein of a 40,92 kDa with two putative cleavage sites, which could origin two processed protein forms of 32,86 and 26,09 kDa. As ebaf mRNA levels were higher during the fourth day of pregnancy, compared to the estrous cycle and other days of pregnancy, we studied samples at this stage. A western blot assay with 100 ug of oviduct total protein was performed, a positive (testicle), a negative (bowel) and a secondary antibody controls were included. We found two bands one of 41 KDa (precursor protein) and other of 26 kDa (processed form). We localized Ebaf in the stroma of the oviduct and mesosalpinx (the peritoneum of the broad ligament) by immunohistochemical techniques. Taken into account these results we suggest that Ebaf is synthesized and processed in the oviduct were it might have a relevant function during the early stages of pregnancy.