Study of in vitro maturation of canine oocytes in a bi-phasic system using LH and progesterone

• Grant number: 10/09231-7
• Support Opportunities: Regular Research Grants
• Start date: July 01, 2010
• End date: June 30, 2012
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Wilter Ricardo Russiano Vicente
• Grantee: Wilter Ricardo Russiano Vicente
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil

Abstract

The use of assisted reproduction techniques such as in vitro maturation and fertilization in canids aims the preservation of threatened canids genome. In addition, recent studies have been performed to define a specific culture media for the species, as well as to enhance maturation ratio of canine oocytes. Nevertheless, the goal of the present study is to evaluate the effects of a bi-phasic system using LH and progesterone in different stages of the in vitro culture of canine oocytes. Thirty canine females, with 1 to 7 years old will be grouped in Follicular Phase (FF) and anestrous (A) groups. Ovaries will be harvested by ovariohysterectomy (OHE) and the oocytes classified according to Hewitt & England (1998). Only cumulus-oocyte grade I (COC) with diameter greater than 120 ¼m will be selected. The oocytes of each phase will be arranged in 4 groups: GI (control: TCM -199), GII (control and progesterone after 48 hours of maturation), GIII (control and LH in the first 48 hours of maturation) e GIV (control and LH in the first 48 hours and progesterone after 48 hours). The samples will be evaluated before (time 0), and after 96 hours of the maturation process. Morphology of the oocytes and cumulus cells will be analyzed by scanning (SEM) and transmission electronic microscopy (TEM). Data will be evaluated by the chi-square test to compare means (Pe0.05%). (AU)