Single Nucleotide Polymorphisms Of ESR,FSHβ And Leptin Genes In Swine And Their Correlations With Follicular Cysts

Ovarian follicular cysts are one of the common reproductive disturbance in sows,which can result in the infertility of swine.They could induce female a nimal have sustained,frequent,brief and irregular estrus.This disease has caus ed huge impediment of the development of pig aquaculture.Now,as the molec ular biology develops,the studies of the detection of swine follicular cysts mar ker genes receive more and more attentions.ESR、FSHβ and Leptin can play i mportant role on the growth of sow ovary via regulating each steps of hypot halamic-pituitary-ovarianaxis.Estrogen receptors(ESR)are one of the steroid receptor family,which can participate in the regulation of female mammals reproductive activities,and effect the expression and regulation of female animal gonadal tissue genes.Thus ESR can affect the animal secondary sex characteristics,reproductive cycle,fertility,pregnancy maintenance.The analysis results showed that ESR Pvu II polymorphism restriction site have significant effect on the the pig litters and reproductive characters.At present,however,there are no report which analysis the association between ESR Pvu II polymorphic sites and follicular cysts.Follicle-stimulating hormone(FSH)are a kind of glycoprotein,and FSH have an important effect on promoting granular cell develop.It was found that pig FSH gene structure contain a retrotransponson(292bp in length).The polymorphism of the gene locus was caused by the insertion of retrotransponson.After the analysis of FSHβ gene polymorphism site,the FSH gene were the major gene which effect the pig reproductive characters.At present,however,there were no study which select FSH as candidate gene to analysis the association between FSH gene polymorphisms and follicular cysts.Leptin(also known as OB)are a kind of proteohormone and are secreted by adipose cells.Their gene expression product are mainly involved in the regulation of animal development,growth and reproductive function.It has been found that the mutation of T3469 C locus had certain influence on the pig reproductive performance.However,there were no study which select Leptin as candidate gene to analysis the association between Leptin gene polymorphisms and follicular cysts.In this experiment,we choose estrogen receptors(ESR),follicle stimulating hormone β subunit gene(FSHβ)and(Leptin)gene as research sample.We try to screen gene polymorphisms sites of estrogen receptors(ESR),follicle stimulating hormone β subunit gene(FSHβ)and(Leptin)gene by normal and cyst ovary collection,extracting genome DNA,direct PCR or PCR-RFLP methods.Then we try to analysis the association between ESR,FSHβ,Leptin gene polymorphisms and follicular cysts,and find the molecular markers of follicular cysts.The main contents are as follows:(1)The detection results of gene polymorphisms showed that 4 polymorphic sites were detected within 3 genes,which were located in the ESR gene intron A > G mutation,FSH β gene intron 5 ’regulatory region T>C mutation,FSHβ gene intron C>T mutation and Leptin genet exon C >T mutation.(2)The analysis result of polymorphism information content,heterozygosity and effective allele showed that ESR Pvu II polymorphism restriction site showed moderate polymorphism in the two groups(0.25<PIC<0.5),and ESR genotype distribution were deviated from the Hardy-Weinberg equilibrium in cyst group(p<0.05),but ESR genotype distribution were in accordance with the Hardy-Weinberg equilibrium in healthy group(p>0.05).FSHβ 5’ regulatory region polymorphism site showed low polymorphism in follicular cysts group(PIC<0.25),and showed moderate polymorphism in healthy group(0.25<PIC<0.5).The genotype distribution of FSHβ 5’ regulatory region polymorphism sites in two groups were all deviated from the HardyWeinberg equilibrium(p<0.05);The HaeⅢ polymorphism restriction site of FSH β showed low polymorphism(PIC< 0.25)in two groups,and the polymorphism level was low.The genotype distribution of this polymorphism site was in accordance with the Hardy-Weinberg equilibrium(p>0.05);The Hinf I polymorphism restriction site of Leptin gene showed low polymorphism(PIC<0.25)in two groups.The genotype distribution of this polymorphism site were in accordance with the Hardy-Weinberg equilibrium in follicular cyst group,but deviated from the Hardy-Weinberg equilibrium(p<0.05)in health group.(3)The analysis of ESR FSHβ and Leptin genotype distribution was carry out by analysis software.The results showed that ESR AA and AB genotypes had significantly difference between healthy group and follicular cyst group(p<0.05),which had relation to the formation of follicular cysts;The genotype distribution of FSH and Leptin gene had no significant difference between 2 groups(p>0.05),which had no relation to the formation of follicular cysts.ESR gene can be used as marker gene for follicular cyst screening,and laid the foundation for researching new techniques of increasing sow fecundity.