UTILIZATION OF THE IMMUNOHISTOCHEMISTRY IN A CELL MIGRATION IN GESTATIONS DERIVED BY CLONED BOVINE CONCEPTUSES THAT EXPRESS THE GREEN FLUORESCENT PROTEIN (GFP)

• Grant number: 07/08010-4
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: December 01, 2007
• End date: November 30, 2008
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Flávia Thomaz Verechia Pereira
• Grantee: Rodrigo da Silva Nunes Barreto
• Host Institution: Universidade Estadual Paulista (UNESP). Campus Experimental de Dracena. Dracena , SP, Brazil

Abstract

Actually the nuclear transfer is the most utilized technique in cloning of young and adult individual, however the success rate still low and associated with placental alterations caused by epigenetic deregulation of imprinted genes in embryogenesis, due to incomplete nuclear reprogramming. For better efficiency of nuclear transfer, should be required more accurate methods for selection of the nucleus donor cells and reconstructed embryos, so we have been using the green fluorescent protein for select transgenic cloned embryos that will use in this work for accompanying, in bovines, the migration of trophoblastic giant cells for uterine epithelium. Associated with this migratory event we have proteoglicans - integrins related with adhesion of the cellular matrix, motility and cellular differentiation - and cellular matrix components - collagen IV - that will be mapped. Thus, we will be utilized transgenic cloned bovine concepts with green fluorescent protein expression, concepts from in vitro fertilization and from natural stud/artificial insemination, part of these samples are already colleted. These samples will be preserved and processed according with specific immunohistochesmitry protocols, will be documented by light and fluorescence microscope and analyzed statistically.