Stress indicator genes expression pattern in bovine blastocysts produced by IVF and SCNT

CÁNEPA MJ, KAISER G, MUCCI N AND MUTTO A

Phoenix

Congreso; IETS; 2012

IETS

Reproductive biotechnologies, such as in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT) enable to improve reproductive efficiency of animals. However, the birth rate from in vitro derived embryos still lags behind their in vivo counterparts. It is critical to develop an accurate evaluation and prediction system of embryo competence, for commercial purposes and for scientific research. Previous works have demonstrated that in vitro culture systems induce alterations in the relative abundance (RA) of diverse transcripts compromising embryo quality. The aim of this work was to analyze the RA of a set of genes involved in cellular stress (HSP70), endoplasmic reticulum (ER) stress (BiP and PSMB5) and apoptosis (Bax and Caspase-3), in bovine blastocysts produced by IVF, SCNT and their in vivo counterparts. Poly(A) RNA was isolated from three pools of 10 blastocysts per treatment and analyzed by Real-Time RT-PCR. The results were normalized to the expression of the endogenous control (GAPDH). Data of mRNA expression was analyzed using the InfoStat software (Universidad Nacional de Córdoba, Argentina). One-way Analysis of Variance ANOVA, followed by Tukey multiple comparison test, was used for the analysis of differences in mRNA expression assayed by quantitative RT-PCR. Differences of p