Polymorphism Analysis Of Inha Gene And In Su Huai Pig And The Development Of Ovary In Piglets

Inhibin alpha subunit and beta A subunit forms inhibin A,while inhibin B contains inhibin alpha subunit and beta B subunit.Due to inhibin alpha subunit has glycosylation site,so inhibin alpha subunit is a key factor in the formation of inhibin.Inhibin A is one of the meof mbers the TGF family,which plays an important role in the regulation of ovarian development and hormone secretion.In mammalian peripheral blood,the formation and secretion of FSH is regulated by INHA/Activin/Fsp.INHA can also regulate the growth/proliferation/differentiation of granulose cells and thecia cells through paracrine and autocrine patterns,and then it affects the development of follicles.INHA also attends the formation and apoptosis of corpus luteum.we use the immune agents of inha in mammalian molecular breeding,especially in ruminant breeding.INHA also plays an important role in the progress of polycystic ovary syndrome,premature ovarian failure and ovarian cancer,and many ovarian diseases are associated with the disorder of INHA expression.With the development of modern civilization,environment pollution becomes more and more serious.In the process of growth,the piglets are easily disturbed by the external environment,especially the environmental endocrine disruptors(EEDs).it has adverse affects on reproduction organs,as premature reproduction/abnormal reproductive organs.The aim of these study are to analyze the genetic polymorphism of inhibin subunit gene and to study the premature ovarian development on its function.The primers were designed according to the gene sequences of GeneBank(Sus scrofa),which had been logged on the internet;We used bioinformatics softwares to analyze the sequences;we analyzed the connection between polymorphism of inha alpha subunit gene and the litter size in Suhuai pigs;Using HE and IHC analyzed the ovary in 60-day-old piglets.The main results were as follows:1.Bioinformatics analysis of 5'UTR region in inhibin alpha subunit geneThe length of the 5'UTR region of inhibin alpha subunit gene was 67bp,and found it had a higher in homology with human/pig/goat.Using the online software Methprimer(http://www.urogene.org)to predict the sequence,we found that there was no CpG island in this sequence.Using online software genomatic(www.genomatic.de.com)to predicted thetranscription factor,results indicated that there were many transcription factors binding 5'UTR region of inhibin alpha gene,such as PLAG1,PRDM5,EKLF,TBX21,CAX6 and so on.2.The connection between the polymorphism of inhibin alpha gene and litter size in Suhuai pigs.Analyzing the single nucleotide polymorphism of inhibin alpha subunit gene in Suhuai pigs,we found a polymorphic loci in in the 5 UTR region(-42G>A),and 4 polymorphic sites in the CDS region(3241C>T,3249A>G,3285G>A and 3294G>A),these four sites can cause amino acid changes.Which were not change the tertiary structure of INHA protein.At the-42G>A locus,G was the dominant allele(frequency 0.553).The correlation analysis showed that the GG genotype of the number of litter was significantly higher than that of type AA.Predicting transcription factors in the 5'UTR region,we found the mutation site could alter the binding of transcription factors,thereby it affected the transcription of inhibin alpha subunits.There was no significant difference between the genotypes at 4 polymorphic sites in CDS area and the litter size of Suhuai pigs.3.Histologic observation and expression patterns of PCNA and Caspase-3 in precocious puberty sowsWe selected 8 pigs randomly and killed them at 60 days old.And then we collected the ovary and uterus to analysis by hematoxylin-eosin staining and immunohistochemistry.The results showed that there is significant difference between the normal group and precocious group in uterine weight(P<0.01).Precocious group has a large number of tertiary follicles,granulosa cells have normal value-added features and produce estrogen in antral follicle.Precocious group's ovary decreased many primordial follicles compared normal group,it may cause dysplasia in ovary and uterus.Thus,premature ovarian development would affect the normal function.