背景:在中国的淡水河流中广泛分布。类似于Medaka(Oryziaslatipes),中国的石竹具有体积小的特点,快速生殖周期,适应性强,这使其适合作为基础生物学和环境毒理学研究的模式生物。中国的田园诗表现出明显的性二态性。然而,由于缺乏完整的基因组信息,在中国,性别决定和分化相关基因的调控仍不清楚。
方法:用PCR方法克隆了中国medakadmrt1(Osdmrt1),产生过表达Osdmrt1的medaka[Tg(CMV:Osdmrt1)]转基因个体,以研究Osdmrt1在性别决定中的作用。Western印迹用于验证Osdmrt1整合到medaka基因组中。组织切片和HE染色用于鉴定Tg(CMV:Osdmrt1)生理性别和表型。qRT-PCR用于分析性腺特异性基因的表达。
结果:克隆并鉴定了Osdmrt1,与mrt1具有相似的进化关系。Tg(CMV:Osdmrt1)在F2代表现出从女性到男性的部分性别逆转,基因女性个体发育睾丸并产生功能性精子。此外,转基因雌性的第二性征也改变为雄性。
结论:中国medakadmrt1基因可以将medaka中的女性转化为男性。
结论:这些结果不仅阐明了中国medakadmrt1的功能,而且为通过转基因技术研究模型鱼中经济鱼功能基因的功能积累了知识。
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.
CONCLUSIONS: The Chinese medaka dmrt1 gene could convert females to males in medaka.
CONCLUSIONS: 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.