The novel porcine gene early growth response 4 (Egr4) is differentially expressed in the ovaries of Erhualian and Pietrain pigs

Abstract

The early growth response 4 (Egr4) gene plays a critical role in human and mouse fertility. In the present study, Affymetrix microarray gene chips were used to evaluate differential gene expression in the ovaries between Erhualian and Pietrain pigs. In all, 487 and 573 transcripts were identified with significantly higher and lower expression, respectively, in Erhualian compared with Pietrain sows. The Egr4 gene, one of the differentially expressed genes, was cloned and its genomic structure was analysed. Egr4 expression is increased 120-fold in ovaries from Erhualian sows. The full-length cDNA of porcine Egr4 was obtained by in silico cloning and 5' rapid amplification of cDNA ends. The gene consists of two exons and its predicted protein contains a Cys2His2 zinc finger structure. The porcine transcript is alternatively spliced by exon sequence deletion, producing two different mRNAs differing at the 5' end of Exon 2. Egr4 transcripts were detected in the central nervous system, including the cerebrum, cerebellum, hypothalamus and pituitary gland, and were highly expressed in the ovary. The Egr4 gene was evaluated as a candidate gene for porcine reproductivity. To investigate the role of Egr4 in the ovary, Egr4 was knocked down using short interference (si) RNA in porcine granulosa cells. Knockdown of Egr4 using siRNA effectively inhibited Egr4 mRNA and protein expression and knockdown significantly affected the expression of Bax, P450arom, P450scc, Egr1, Egr2, and Egr3. In conclusion, these observations establish an important role for Egr4 in the porcine ovary.