梅山与长大母猪下丘脑-垂体-卵巢轴Kiss1和GPR54基因的表达差异

研究早熟与晚熟品种母猪下丘脑-垂体-卵巢轴Kiss1和GPR54基因的表达差异。选用8头梅山母猪与12头长大(LY)母猪为研究对象,梅山母猪于70 d和100 d屠宰,LY母猪于70、100 d和199 d屠宰,收集血清及下丘脑、垂体、卵巢组织样品。ELISA检测血清瘦素(Leptin)和雌二醇(E2)水平,PCR克隆梅山与LY母猪Kiss1基因编码序列,实时荧光定量PCR检测母猪下丘脑、垂体、卵巢组织Kiss1和GPR54基因表达水平。结果表明:梅山与LY母猪Kiss1基因编码序列相似性为100%;梅山与LY母猪初情期下丘脑Kiss1基因表达量极显著高于垂体与卵巢(P<0.01),卵巢GPR54基因表达水平显著高于下丘脑与垂体(P<0.05)。梅山母猪下丘脑-垂体-卵巢轴Kiss1基因表达水平都显著高于相同日龄和初情期LY母猪(P<0.05),梅山母猪血清Leptin水平极显著高于相同日龄LY母猪(P<0.01)。而E2水平显著高于100 d与初情期LY母猪(P<0.01)。Leptin与下丘脑Kiss1和GPR54基因表达呈显著正相关(P<0.05),而E2仅与下丘脑Kiss1基因表达有极显著正相关关系(P<0.01);梅山与LY母猪Kiss1基因编码区序列相似性为100%,梅山母猪下丘脑-垂体-卵巢轴上kiss1基因表达量较LY母猪高,主要原因可能是初情日龄早,下丘脑Kiss1基因表达水平的高低与血清Leptin和E2浓度密切相关。

The Difference of Kiss1 and GPR54 Gene Expressions in Hypothalamus-Pituitary-Ovary Axis of Meishan and LY Breed Gilts

To investigate the difference between kiss1 and GPR54 gene expressions in hypothalamus-pituitary-ovary axis in the pre-maturity and late-maturity breed gilts.8 Meishan(MS) gilts and 12 Large white×Yorkshire(LY) breed gilts were selected and sacrificed at day 70,100 and day 70,100,199,respectively.Serum,hypothalamus,pituitary,ovary samples were collected for analysis.The concentrations of serum leptin and estradiol were measured by enzyme-linked immunosorbent assay.The coding sequences of Kiss1 gene in MS and LY gilts were cloned by using TA-cloning,and the expression of kiss1 and GPR54 gene were detected by real-time PCR.The coding sequences of Kiss1 gene in MS and LY gilts have a similarity of 100%.MS and LY gilts hypothalamus Kiss1 gene expression were significantly higher than pituitary and ovary in puberty(P <0.01),while GPR54 gene expression in ovary was significantly higher than that in hypothalamus and pituitary at the same time(P <0.05).The expression of Kiss1 gene in hypothalamus-pituitary-ovary axis in MS gilts was significantly higher than that in LY gilts at the same age and puberty,while GPR54 expression was not significant(P> 0.05).The concentration of serum Leptin in MS gilts was significantly higher than that at LY gilts at the same age(P <0.01),while E2 levels was significantly higher than that of LY gilts at 100 d and puberty(P <0.01).Serum leptin was significantly positively correlated to the expressions of Kiss1 and GPR54 gene in hypothalamus at puberty(P <0.05),but E2 was significantly positively correlated with Kiss1 gene expression in hypothalamic only(P <0.01),In conclusion,the coding sequences of Kiss1 gene in MS and LY gilts have a similarity of 100%.MS gilts show early puberty onsets age than LY gilts,which were probably determined by a higher Kiss1 gene expression in hypothalamus-pituitary-ovary axis,while hypothalamic Kiss1 expression were closely positively related to serum Leptin and E2 concentrations.