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采用半定量PCR的方法,分别从时空表达、温度应激调控和血清雌二醇(Estradiol-17β,E2)水平3个方面,共同研究雌性黄颡鱼Sox9基因的功能。结果表明,Sox9a1基因在雌性黄颡鱼各组织中存在广泛的表达,而Sox9a2则在脑和卵巢中有高表达(脑>卵巢),胃中表达微弱。在卵巢年周期发育中,卵巢中Sox9a1在Ⅲ期时达到高峰,Sox9a2则在Ⅴ期时达到峰值;脑中Sox9a1和Sox9a2表达的最大值均在Ⅱ期,后随着卵巢的发育表达水平显著下降。在热应激调控试验中,卵巢中Sox9a1虽然对热应激存在响应,但并没有出现E2相一致的变化趋势,而Sox9a2的变化趋势与血浆中E2相同。虽然脑中Sox9a1和Sox9a2在卵巢年周期发育的表达变化出现下降的趋势,但在热应激条件下,脑中Sox9a1和Sox9a2在Ⅱ期与E2的变化相反,而Ⅲ期到Ⅵ期与E2的变化相一致。该实验结果表明,卵巢中Sox9a1可能不参与卵巢的年周期发育,Sox9a2则参与卵巢发育的调控和E2的合成;在脑中,我们推测热应激可能改变了Sox9与雌激素的负调控关系,但其机理需要更深入的研究。
Semi-quantitative PCR method was used to study the function of Sox9 gene from three aspects of spatiotemporal expression, temperature stress regulation and the level of estradiol-17β (E2). The results showed that Sox9a1 gene was widely expressed in various tissues of the female yellow soleus, while Sox9a2 was highly expressed in the brain and ovary (brain> ovary). The expression of Sox9a1 was weak in the stomach. During ovarian cycle development, Sox9a1 peaked in stage Ⅲ and Sox9a2 peaked in stage Ⅴ. The maximum expression of Sox9a1 and Sox9a2 in stage Ⅱ was significantly decreased in ovary with the development of ovary . In the regulation of heat stress, ovary Sox9a1 although there is a response to heat stress, but did not appear consistent trend of E2, and Sox9a2 trends and E2 in plasma the same. Although the expression of Sox9a1 and Sox9a2 in the brain decreased during the ovarian cycle development, under the condition of heat stress, Sox9a1 and Sox9a2 in the brain changed oppositely to E2 in stage Ⅱ, whereas in stage Ⅲ to Ⅵ and E2 Change consistent. The experimental results show that ovary Sox9a1 may not participate in ovarian cycle development, Sox9a2 is involved in the regulation of ovarian development and E2 synthesis; in the brain, we speculated that heat stress may change the negative regulation of Sox9 and estrogen, But its mechanism needs more research.