Expression of caspase-3, caspase-9, GDF-9 and IGF-1 genes in ovine oocytes cultured with optimum and elevated doses of Amphiregulin, Neuregulin-1 and Tumour necrosis factor-α in-vitro

Abstract

The present study was conducted to evaluate the expression of caspase-3, caspase-9, growth differentiation factor-9 (GDF-9) and insulin growth factor-1 (IGF-1) genes in oocytes cultured in vitro with optimum and elevated doses of amphiregulin (50 ng and 150 ng), neuregulin-1 (25 ng and 150 ng) and tumor necrosis factor-α (25 ng and 150 ng) during in vitro maturation of oocytes based on the results of effects of AREG or NRG-1 or TNF-α concentration on maturation of oocytes which caused significant effect, were selected for gene expression studies along with the basal/control level. Total RNA was extracted from in vitro matured oocytes using Trizol method and Real-time reverse transcription polymerase chain reaction was used to evaluate the expression of genes. There was an upregulation of caspase-3 at 150 ng of TNF-α, caspase-9 at 50 ng of AREG, GDF-9 at 150 ng of AREG and IGF-1 at 150 ng of AREG and 25 ng of TNF-α. These results suggested that AREG at elevated dose and TNF-α at optimum dose enhanced the expression of GDF-9 and IGF-1, while the presence of elevated dose of TNF-α and optimum dose of AREG activated caspase-3 and caspase-9, respectively in oocytes cultured in vitro.