Ghrelin is an important signaling molecule linking reproductive and energy metabolism. In this study, ghrelin gene of Monopterus albus was cloned. Its structure and function were analysized preliminarily. By Rapid Amp...Ghrelin is an important signaling molecule linking reproductive and energy metabolism. In this study, ghrelin gene of Monopterus albus was cloned. Its structure and function were analysized preliminarily. By Rapid Amplification of cDNA Ends(RACE) technique, full-length cDNA and DNA sequences of ghrelin gene were obtained. The full-length ghrelin cDNA(GenBank accession no. JX122807) was 552 bp long, containing a 115 bp 5'-untranslated region, a 324 bp open reading frame and a 113 bp 3'-untranslated region. The full-length ghrelin DNA was 1 323 bp, consisting of three introns and four exons. The exon/intron junction sequences conformed to the GT/AG rule. Three introns were 594, 84 and 93 bp in length, respectively; four exons were229, 78, 112 and 133 bp in length, respectively. The results of amino acid sequence analysis showed that the deduced propreghrelin sequence of M. albus contained a signal peptide(SP) consisting of 22 amino acid residues, a mature peptide(MP)consisting of 19 amino acid residues and a C-terminal amino acid residue. Among them, the third amino acid of MP was serine(Ser^3) as the site for N-acylation and N-deacetylation reactions; the C-terminal amino acid residue sequence might contain a peptide hormone obestatin, which is a physiological antagonist of mature Ghrelin peptide. The homology and phylogenic relationships analyses of amino acid sequences suggested that propreghrelin of M. albus had high similarity to those of several Perciformes fishes; the propreghrelins of M. albus, Perciformes and Heterosomata fishes were clustered into a subgroup. The high conservatism of the gene structure and the amino acid sequences indicated that Ghrelin exerts important physiological functions and plays similar physiological mechanisms in vertebrates.展开更多
目的:探讨Ghrelin基因在卵巢颗粒细胞中的表达及其与多囊卵巢综合征(polycystic ovarian syndrome,PCOS)的关系。方法:选择47例在重庆医科大学辅助生育中心接受体外受精-胚胎移植治疗的不孕妇女,其中PCOS组11例,对照(Non-PCOS)组36例,采...目的:探讨Ghrelin基因在卵巢颗粒细胞中的表达及其与多囊卵巢综合征(polycystic ovarian syndrome,PCOS)的关系。方法:选择47例在重庆医科大学辅助生育中心接受体外受精-胚胎移植治疗的不孕妇女,其中PCOS组11例,对照(Non-PCOS)组36例,采用real-time PCR及免疫印迹法分别测定2组妇女卵巢颗粒细胞中Ghrelin m RNA和蛋白的表达水平。结果:PCOS妇女颗粒细胞中Ghrelin m RNA和蛋白的表达均低于Non-PCOS妇女(0.926±0.206 vs.1.137±0.105,t=2.73,P=0.015;0.647±0.052 vs.0.937±0.038,t=7.82,P=0.001)。结论:PCOS妇女卵巢颗粒细胞的Ghrelin m RNA和蛋白表达水平明显低于Non-PCOS妇女,可能与卵泡发育障碍相关。展开更多
基金Supported by Natural Science Foundation of Hubei Province(2013CFB393)Open Fund of Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilizations,Ministry of Agriculture(KF201307)+2 种基金National Pillar Program of China(2013BAD20B06)Pillar Program of Hubei Province(2015BBA235)Program for Outstanding Young and Middle-aged Scientific Innovation Team in Colleges and Universities of Hubei Province(T201503)
文摘Ghrelin is an important signaling molecule linking reproductive and energy metabolism. In this study, ghrelin gene of Monopterus albus was cloned. Its structure and function were analysized preliminarily. By Rapid Amplification of cDNA Ends(RACE) technique, full-length cDNA and DNA sequences of ghrelin gene were obtained. The full-length ghrelin cDNA(GenBank accession no. JX122807) was 552 bp long, containing a 115 bp 5'-untranslated region, a 324 bp open reading frame and a 113 bp 3'-untranslated region. The full-length ghrelin DNA was 1 323 bp, consisting of three introns and four exons. The exon/intron junction sequences conformed to the GT/AG rule. Three introns were 594, 84 and 93 bp in length, respectively; four exons were229, 78, 112 and 133 bp in length, respectively. The results of amino acid sequence analysis showed that the deduced propreghrelin sequence of M. albus contained a signal peptide(SP) consisting of 22 amino acid residues, a mature peptide(MP)consisting of 19 amino acid residues and a C-terminal amino acid residue. Among them, the third amino acid of MP was serine(Ser^3) as the site for N-acylation and N-deacetylation reactions; the C-terminal amino acid residue sequence might contain a peptide hormone obestatin, which is a physiological antagonist of mature Ghrelin peptide. The homology and phylogenic relationships analyses of amino acid sequences suggested that propreghrelin of M. albus had high similarity to those of several Perciformes fishes; the propreghrelins of M. albus, Perciformes and Heterosomata fishes were clustered into a subgroup. The high conservatism of the gene structure and the amino acid sequences indicated that Ghrelin exerts important physiological functions and plays similar physiological mechanisms in vertebrates.
文摘目的:探讨Ghrelin基因在卵巢颗粒细胞中的表达及其与多囊卵巢综合征(polycystic ovarian syndrome,PCOS)的关系。方法:选择47例在重庆医科大学辅助生育中心接受体外受精-胚胎移植治疗的不孕妇女,其中PCOS组11例,对照(Non-PCOS)组36例,采用real-time PCR及免疫印迹法分别测定2组妇女卵巢颗粒细胞中Ghrelin m RNA和蛋白的表达水平。结果:PCOS妇女颗粒细胞中Ghrelin m RNA和蛋白的表达均低于Non-PCOS妇女(0.926±0.206 vs.1.137±0.105,t=2.73,P=0.015;0.647±0.052 vs.0.937±0.038,t=7.82,P=0.001)。结论:PCOS妇女卵巢颗粒细胞的Ghrelin m RNA和蛋白表达水平明显低于Non-PCOS妇女,可能与卵泡发育障碍相关。
