| 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 |
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| 引用本文： | 黄嘉慧,郭昱嵩,杜娟,董忠典,王中铎.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[J].水产学报,2021,45(12):1965-1972. |
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| 作者姓名： | 黄嘉慧 郭昱嵩 杜娟 董忠典 王中铎 |
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| 作者单位： | 广东海洋大学水产学院,广东海洋大学水产学院,广东海洋大学水产学院,广东海洋大学水产学院,广东海洋大学水产学院 |
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| 基金项目： | 国家自然科学(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)；[Supported by National Natural Science Foundation of China (No.41806195, 31972794); |
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| 摘 要： | 体表色素具有保护鱼类免受紫外线辐照伤害等重要功能。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。为了检验这一假说，本研究以相同条件下人工繁育的不同纬度的弓背青鳉(Oryzias curvinotus)群体（饶平、高桥、三亚）后代（F6）为材料，使用荧光倒置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（6-10，12，14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%；相应地，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代胚胎样本的色素细胞发生相关基因qRT-PCR检测确定：tyr表达水平从6肌节期开始上调，至10肌节期趋于平稳；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基因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，本研究支持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、pax3等基因的有序表达的调控，且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。
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| 关 键 词： | 弓背青鳉 色素细胞发生 早期胚胎 群体分化 |
| 收稿时间： | 2020/11/9 0:00:00 |
| 修稿时间： | 2021/3/6 0:00:00 |
Development of pigment cells and analysis of the expression of related genes in the early embryo of Oryzias curvinotus |
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| HUANG Jiahui,GUO Yusong,DU Juan,DONG Zhongdian,WANG Zhongduo.Development of pigment cells and analysis of the expression of related genes in the early embryo of Oryzias curvinotus[J].Journal of Fisheries of China,2021,45(12):1965-1972. |
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| Authors: | HUANG Jiahui GUO Yusong DU Juan DONG Zhongdian WANG Zhongduo |
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| Institution: | Guangdong Ocean University,Guangdong Ocean University,Guangdong Ocean University,Guangdong Ocean University,Guangdong Ocean University |
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| Abstract: | 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 (F6) 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 embryos were concentrated in 7 and 11 somite stages, accounting for 47.7% and 42.1%, respectively; And the melanophores and iridophores of the embryo in Sanya embryos were concentrated in 8 and 12 somites stage, accounting for 44.0% and 62.2%. qRT-PCR analysis of Gaoqiao embryos confirmed that the expression of tyr, the rate-limiting enzyme of melanin formation, was up-regulated from the 7 to 9 somites stage, and then tended to be stable; And the sox10 and pax3 genes, which inhibit the formation pathway of melanin and initiate the differentiation of iridescent cells, showed the single peak expression pattern from the 10 to 14 somites stage as the downstream alk genes. In conclusion, this study supports that the pattern of melanin and iridescent pigments in the early development stage of O. curvinotus is consistent with the classical pathway of fish, which is regulated by the orderly expression of tyr, alk, sox10, pax3 genes, and the iridescent cells in the low latitude population generally occur later, which supports the existence of population genetic differentiation, but the specific mechanism remains to be studied in the future. |
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| Keywords: | Oryzias curvinotus pigment cells embryo genetic differentiation |
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