不同倍性鱼的gnih和gnihr3差异表达分析  

Comparative analysis of expression of gnih and gnihr3 genes in different ploidy fishes

作　　者：袁柳娇 黄露[1,2] 范思宇 李胜男 周天[1,2] 赵如榕 陶敏 刘少军[1,2] YUAN Liujiao;HUANG Lu;FAN Siyu;LI Shengnan;ZHOU Tian;ZHAO Rurong;TAO Min;LIU Shaojun(State Laboratory of Developmental Biology of Freshwater Fish,Hunan Normal University,Changsha 410081,China;Engineering Research Center of Polyploid Fish Reproduction and Breeding of the State Education Ministry,College of Life Sciences,Hunan Normal University,Changsha 410081,China)

机构地区：[1]湖南师范大学省部共建淡水鱼类发育生物学国家重点实验室,湖南长沙410081 [2]湖南师范大学生命科学学院,多倍体鱼繁殖及育种技术教育部工程研究中心,湖南长沙410081

出　　处：《水产学报》2020年第10期1585-1598,共14页Journal of Fisheries of China

基　　金：国家自然科学基金(31872551,31730098,31702328);湖南省杰出青年科学基金(2020JJ2022);现代农业体系建设专项资金(CARS-45);111计划(D20007)。

摘　　要：为研究促性腺激素抑制激素(gonadotropin inhibitory hormone,GnIH)及其受体GnIHR在不同倍性鲫鲤生殖发育过程中的作用,实验通过PCR和RACE技术获得了二倍体红鲫和异源三倍体鲫的gnih、gnihr3基因cDNA全长。结果显示,2种鱼gnih基因编码的蛋白序列均含有GnIH肽典型的LPXRF标志,gnihr3基因编码的蛋白为含有7个跨膜区的G蛋白偶联受体。采用实时荧光定量PCR分析了2种鱼的gnih、gnihr3基因mRNA在不同组织中的表达情况,结果显示gnih基因主要在下丘脑中高表达;gnihr3基因在下丘脑-垂体-性腺(HPG)轴中都有不同程度的表达;此外,无论是非繁殖期或繁殖期,gnih基因在红鲫下丘脑中的表达量均高于异源三倍体鲫;同时,gnihr3基因在红鲫下丘脑和垂体中的表达量均高于异源三倍体鲫,但在卵巢中的表达则后者较高。利用组织原位杂交分析了gnih和gnihr3基因在2种鱼下丘脑和垂体的组织定位情况,结果发现,gnih基因在2种鱼脑中Npp、Npo、NLT区均有明显杂交信号;gnihr3基因主要定位在这2种鱼垂体的PPD、RPD区,NH和PI区杂交信号较弱;同时,gnih和gnihr3基因在异源三倍体鲫中的杂交信号比红鲫弱。研究表明,在下丘脑大量表达的GnIH,结合广泛分布于HPG轴的受体GnIHR,并通过多条途径参与调控性腺发育。gnih、gnihr3 mRNA在2鱼组织中的表达存在差异性,从分子水平提供了异源三倍体鱼不育的证据,在鱼类繁殖生物学和遗传育种方面具有重要意义。In order to study gonadotropin inhibitory hormone(GnIH)and its receptor encoded by gnih and gnihr genes in gonadal development of different ploidy cyprinid fish,molecular cloning and comparative expression patterns of gnih and gnihr3 genes in diploid red crucian carp and allotriploid crucian carp were carried out.By PCR and RACE technology,the full-length cDNA sequences of gnih and gnihr3 genes of diploid red crucian carp and allotriploid crucian carp were obtained.Our data showed the amino acid sequences of GnIH encoded by gnih gene of red crucian carp and allotriploid crucian carp both contained the typical LPXRF marker of GnIH peptide.GnIHR3 encoded by gnihr3 gene was a G-protein coupled receptor with seven transmembrane domains.Real-time PCR was used to analyze the expressions of gnih and gnihr3 genes in different tissues of different ploidy Cyprinids.The results showed that the expression level of gnih gene in the hypothalamus of red crucian carp was higher than that of allotriploid crucian carp;the gnihr3 gene was expressed differently on the hypothalamic–pituitary–gonad axis(HPG).Nevertheless,it was noteworthy that regardless of non-breeding or breeding season the expression of gnih in the hypothalamus from red crucian carp was higher than that of allotriploid crucian carp.Simultaneously,the expression level of gnihr3 gene in hypothalamus and pituitary of red crucian carp was higher than that of allotriploid crucian carp,but the expression of gnihr3 in the ovary of allotriploid crucian carp was higher than that of red crucian carp.Through in situ hybridization,the locations of gnih and gnihr3 genes in brain or pituitary of different ploidy fishes were analyzed.Strong hybridization signals of gnih could be detected in Npp,Npo,NLT in the brain of red crucian carp and allotriploid crucian carp.Strong hybridization signals of gnihr3 were located mainly in the PPD and RPD regions of the pituitary,and weak hybridization signals were distributed in NH and PI regions.Meanwhile,the signal of gnihr3 hybridizatio

关 键 词：异源三倍体鲫 GNIH gnihr3 MRNA表达 不育 

分 类 号：S917.4[农业科学—水产科学]