弓背青鳉早期胚胎色素细胞发生及相关基因表达规律分析  

Development of pigment cells and analysis of the expression of related genes in the early embryo of Oryzias curvinotus

作　　者：黄嘉慧 郭昱嵩[1] 杜娟 董忠典 王中铎[1,2] HUANG Jiahui;GUO Yusong;DU Juan;DONG Zhongdian;WANG Zhongduo(Key Laboratory of Aquaculture in South China Sea for Aquatic Economic Animal of Guangdong Higher Education Institutes,Fisheries College,Guangdong Ocean University,Zhanjiang 524088,China;State Key Laboratory of Development Biology of Freshwater Fish,College of Life Sciences,Hunan Normal University,Changsha 410081,China)

机构地区：[1]广东海洋大学水产学院,南海水产经济动物增养殖重点实验室,广东湛江524088 [2]湖南师范大学生命科学学院,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

出　　处：《水产学报》2021年第12期1965-1972,共8页Journal of Fisheries of China

基　　金：国家自然科学基金(41806195,31972794);2019年度创新强校工程项目(230419069,230419055);省部共建淡水鱼类发育生物学国家重点实验室开放课题(2020KF004);粤桂联合基金(2020A1515410009)。

摘　　要：为了检验"依照自然选择理论,不同群体的体色形成过程可能受到光照等环境因素的选择作用而发生遗传分化"这一假说,实验以相同条件下人工繁育的不同纬度的弓背青鳉群体(饶平、高桥、三亚)后代(F;)为材料,使用体式荧光显微镜对早期胚胎的黑色素细胞和虹彩色素细胞进行观察统计,并利用实时荧光定量PCR(qRT-PCR)分析4种色素细胞发生关键基因(黑色素形成的限速酶基因tyr、虹彩色素细胞相关基因alk、sox10和pax3)在高桥群体后代早期发育(6~10、12和14肌节期)过程中的表达加以验证。结果表明,高纬度饶平群体早期胚胎的黑色素细胞和虹彩色素细胞分别集中出现于8肌节期和10肌节期,占比为81.9%和52.1%;高桥群体早期胚胎的黑色素细胞和虹彩色素细胞分别集中出现于7和11肌节期,占比47.7%和42.1%;而三亚群体早期胚胎的黑色素细胞和虹彩色素细胞分别集中出现于8和12肌节期,占比44.0%和62.2%。对高桥F6胚胎样本的色素细胞发生相关基因经qRT-PCR检测结果发现,tyr表达水平在7~9肌节期呈现明显上调,随后趋缓;而抑制黑色素形成通路并启动虹彩细胞分化通路的sox10和pax3与下游的虹彩色素细胞相关基因alk在10~14肌节期均呈现一致的单峰表达模式。综上,本研究中弓背青鳉早期发育阶段的黑色素细胞和虹彩色素发生模式与鱼类经典理论相符,受tyr、alk、sox10和pax3等基因的有序表达的调控,证实了发生阶段存在由遗传因素决定的群体分化,低纬度群体的虹彩细胞发生更晚,其具体机制仍待进一步研究。In different species of fish,the number and distribution of pigments are specific,which results in the diversity of body colors.The pigments in fish have lots of functions such as avoiding predators,protecting fish from radiation,and changing their body color for courtship.The diversity of pigments can make fish better adapt to the natural environment.According to the theory of natural selection,the process of formation of pigments of different groups may genetically be differentiated due to the selection effect of environment light and so on.Oryzias curvinotus is a kind of small oviparous fish and its body length is about 2-4 cm.O.curvinotus has strong fertility,a short generation cycle,sensitivity to water quality and environmental changes,and wide adaptability to salinity.O.curvinotus were used to living in the layer water,which is widely distributed in Guangdong coastal areas.Because the embryos of O.curvinotus are transparent,which makes them become the ideal materials for pigment research and gene function research.At present,there have been researches on the resources survey,development,and function of O.curvinotus which is expected to be developed as a model species for monitoring coastal water environment in China.To test the above hypothesis,in this study,different latitude groups(Raoping,Gaoqiao and Sanya)of O.curvinotus were bred under the same conditions,and their progenies(F;)were used as materials.The iridophores and melanophores of embryos of O.curvinotus were observed by stereo fluorescence microscope,in addition,the expression of four main genes(tyr,alk,sox10 and pax3)in the early development of the Gaoqiao population was analyzed by real-time fluorescence quantitative PCR(qRT-PCR),and the observation results were verified.The results of observation showed that the melanophores and iridophores of embryos in high latitude Raoping embryos were concentrated in 8 and 10 somites stages,accounting for 81.9%and 52.1%respectively;Accordingly,the melanophores and iridophores of the embryo in Gaoqiao embr

关 键 词：弓背青鳉 色素细胞发生 早期胚胎 群体分化 

分 类 号：S917.4[农业科学—水产科学]