Screening And Expression Analysis Of Genes Related To Laying Production In Huoyan Geese Ovarian Tissue

In this study, Liaoning Huoyan geese were used as test object. First, total RNA were extracted from laying period and ceased period ovarian tissue, then sequenced using Illumina hiseq 2000 high throughout sequencing technology platform to screen and anlyze function of the differentially expressed transcripts by the biological information tool. Second, the partial fragments of StAR, CYP11Aland CYP19Algenes, which related to the synthesis and secretion of ovarian reproductive hormones were amplified. At last, the expression variations of those three genes in ovarian tissues of Huoyan goose during egg-laying cycle were tested using real-time fluorescence quantitative PCR and Western blotting methods. Our results are listed as follows:(1)688 significantly differentially expressed genes composed of 344 up-regulated and 344 down-regulated genes were successfully screened and identified, which including the genes involved in the steroid hormone synthesis, gonadotropin releasing hormone signaling pathway, oocyte meiosis, progesterone mediated oocyte maturation, steroid synthesis, calcium signal transduction pathway.(2) The fragment of StAR, CYP11A1 and CYP19A1 genes for Huoyan goose were successfully obtained with the size 449 bp,272 bp and 498 bp. Similarity comparison showed that the StAR gene of Huoyan goose has the 99% and 94% similarity with Swan geese and mallard, respectively. The CYP11A1 gene similarity of Huoyan goose with the Swan goose is 99%, mallard is 97%. For CYP19A1 gene, the similarity fo Huoyan goose with Swan goose, mallard’s is 99%, and the chicken is 93%.(3)The change of the mRNA expression of StAR, CYP11A1, CYP19A1 from March, May, August, November is first increased and then decreased, in May the expression was the highest, and compared May with other months,the expression level has a significant difference; in November, StAR and CYP11A1 mRNA expression was the lowest, and CYP19A1 mRNA expression was the lowest in March. The expression of StAR, CYP11A1 and CYP19A1 mRNA were significantly different in different months. The change of protein expression of StAR, CYP11A1, CYP19A1 was consistent with the corresponding mRNA expression levels, from March, May, November.This study showed that the StAR, CYP11A1, CYP19A1 associated with reproductive hormone synthesis in Huoyan goose have the differentially, regular expressed pattern in ovarian tissue of huoyan geese during egg-laying cycle, suggested that they may involved in Huoyan goose egg lay and reproductive function regulation.