The Research Of Pig H3K9Acetylation And Its Regulation To The Fshr Expression

Follicle stimulating hormone (FSH) is one of the glycoprotein gonadotropin.It is synthesized and secreted by the animal anterior pituitary basophilic cells and is an important hormone for the normal gonadal development and reproductive function. FSH plays an important role in mammalian reproduction by interacting with its specific receptor in the gonads.The expression of FSH receptor was affected by the paracrine/autocrine factors or regulatory elements of the promoter region. Therefore, it is very important to study the the FSHR5’flanking region containing regulatory elements. People have cloned the FSHR promoter region in many species, including humans, rats, mice and sheep.But there are few studies on the pig FSHR promoter.Chromosomal DNA was made of the nucleosome. Histone epigenetic modifications on the nucleosomes can affect gene expression, which cause more and moer attention. Previous studies have demonstrated that DNA Methylation can change the gene expression. We can use the chromatin immunoprecipitation techniques to study the relationship between acetylation and FSHR expression regulation. The experimental results are as follows:1Deletion Analysis of the Pig FSHR PromoterAnalysis of FSHR5’region sequences obtained from the chromatin walking found no TATA box, and bioinformatic analysis did not find the CpG islands. Partial of pig FSHR promoter were cloned into the promoterless pGL3-basic vector. Promoter activity was analyzed with truncated promoter function in transiently transfected pig granular cells. The longest construct P1590, containing nucleotides from-1557to+32relative to the transcription start site showed almost2.07-fold higher activity in granular cells compared to the pGL3-basic vector. Further deletion from-1557to-1159led to a3.03-foid higher promoter activity.Further stepwise deletions from-1159to-562obviously decreased compared to the p1185vector. So a construct only containing nucleotides from-1159to-562may be the important promoter regulation regions.2The correlation of H3K9acetylation and FSHR gene expression regulationTo study the relationship between H3K9acetylation and FSHR gene expression in vivo and vitro experiments differently.In vivo experiments,the FSHR promoter acetylation level of healthy follicles was higher than the atretic follicles,while the FSHR expression was high; In vitro experiments, the granulosa cells were treated by FSH and sodium butyrate, and found that FSHR gene expression increased while H3K9acetylation level was high.It indicates that there was a correlation between histone acetylation and gene expression.