Lei Chen , Yan Huang , Qi-Hua Pan , Meng-Yang Wang , Jing-Jie Liang , Tian-Sheng Chen
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引用次数: 0
Abstract
Background
Chinese medaka (Oryzias sinensis) is widely distributed in freshwater rivers in China. Similar to the medaka (Oryzias latipes), Chinese medaka has the characteristics of small size, rapid reproductive cycle, and strong adaptability, which makes it suitable as a model organism for studies in basic biology and environmental toxicology. Chinese medaka exhibits distinct sexual dimorphism. However, due to the lack of complete genomic information, the regulation of sex determination and differentiation-related genes in Chinese medaka remains unclear.
Methods
Chinese medaka dmrt1 (Osdmrt1) was cloned by PCR, and transgenic individuals of medaka [Tg(CMV:Osdmrt1)] overexpressing Osdmrt1 were generated to investigate the role of Osdmrt1 in sex determination. Western blot was used to validate the integration of the Osdmrt1 into the medaka genome. Tissue sectioning and HE staining were used to identify Tg(CMV:Osdmrt1) physiological gender and phenotype. qRT-PCR was used to analyze the expression of gonad-specific genes.
Results
Osdmrt1 was cloned and identified, and it shared similar evolutionary relationships with medaka dmrt1. Tg(CMV:Osdmrt1) exhibited partial sex reversal from female to male in the F2 generation, with genetically female individuals developing testes and producing functional sperm. Additionally, the secondary sexual characteristics of the transgenic females also changed to males.
Conclusion
The Chinese medaka dmrt1 gene could convert females to males in medaka.
General significance: These results not only elucidate the function of Chinese medaka dmrt1, but also accumulate knowledge for studying the function of economically important fish genes in model fish by transgenic technology.
背景中国青鳉广泛分布于中国的淡水河流中。与青鳉相似,中国青鳉具有体型小、繁殖周期快、适应性强等特点,适合作为基础生物学和环境毒理学研究的模式生物。中国青鳉具有明显的性二型。然而,由于缺乏完整的基因组信息,中国青鳉性别决定和分化相关基因的调控仍不清楚:方法:利用PCR技术克隆了中国青鳉的dmrt1(Osdmrt1),并产生了过表达Osdmrt1的转基因青鳉个体[Tg(CMV:Osdmrt1)],以研究Osdmrt1在性别决定中的作用。用 Western blot 验证了 Osdmrt1 与青鳉基因组的整合。用组织切片和HE染色鉴定Tg(CMV:Osdmrt1)的生理性别和表型:结果:克隆并鉴定了Osdmrt1,它与青鳉Dmrt1具有相似的进化关系。Tg(CMV:Osdmrt1)在F2代表现出从雌性到雄性的部分性别逆转,基因上为雌性的个体发育出睾丸并产生功能性精子。此外,转基因雌性个体的第二性征也变为雄性:结论:中国青鳉 dmrt1 基因可使雌性青鳉转为雄性:这些结果不仅阐明了中国青鳉 dmrt1 的功能,而且为利用转基因技术研究经济鱼类功能基因在模式鱼类中的功能积累了知识。
期刊介绍:
BBA General Subjects accepts for submission either original, hypothesis-driven studies or reviews covering subjects in biochemistry and biophysics that are considered to have general interest for a wide audience. Manuscripts with interdisciplinary approaches are especially encouraged.