Characterization of the yolk sac proteins of the Bos indicus bovine embryos

Author(s)

Fabiana Santos Matsumoto

• Document type: Master's Dissertation
• Press: São Paulo.
• Institution: Universidade de São Paulo (USP). Faculdade de Medicina Veterinária e Zootecnia (FMVZ/SBD)
• Defense date: 2007-03-16

Examining board members

Maria Angélica Miglino; Carlos Eduardo Ambrosio; Tatiana Jazedje da Costa Silva

• Advisor: Maria Angélica Miglino

Abstract

In many species of mammals, the yolk sac is one of the embrionary membranes that plays an important role in the embryo´s initial survival, as well as, in the manufacturing of the necessary proteins for its development. In order to identify the proteins: alfafetoprotein, alfa 1 - antitrypsin, and transferrin present in the cow´s embryo´s yolk sac, 17 bovine embryos were collected in different pregnancy periods. This procedure was performed by Western Blot Technique with a polyacrylamide gel electrophoresis, SDS-PAGE, at 6%. Gels following the electrophoresis, where tainted with Comassie blue, and the membranes of Nitrocellulose, following their transference (the proteins that were present in the gel go to the membrane), with Ponceau. Monoclonal Antibody mouse anti human α-fetoprotein, alphafetoprotein mouse monoclonal antibody, transferrin receptor mouse IgG1, and rabbit polyclonal to alpha 1 antitrypsin were used as primary antibodies, and Peroxidase labelled antimouse e Peroxidase labelled antirabbit e anti-mouse IgG- Alkaline Fosfatase as secundary ones. The membrane´s revelation was of the alcaline fosfatase colormetric type and by ECL. The yolk sac was presented well developed until the 50 days of gestation, where to break of this period the involution process well it is characterized. In some of the yolk sac samples we detected the presence of alfafetoprotein, alfa 1- antitrypsin, and transferrin, however, the bands in some specimens (samples) were weak, demonstrating that the antibodies react with the bovine proteins. The fact that weak bands appeared might be related to a weak cross reaction since we are dealing with a non specific antibody. (AU)