咪唑类物质KK-42对日本沼虾头胸甲中MnCP-3表达的影响

Effect of Imidazole Derivative KK-42 on Expression of MnCP-3 in Carapace from Macrobrachium Nipponensis

为了分析咪唑类物质KK-42对日本沼虾表皮蛋白(Macrobrachium nipponense cuticle proteins,MnCPs)基因表达的影响,首次在头胸甲中克隆得到一条基因,通过生物信息学分析确定其属于CPs基因,并将其命名为MnCP-3.采用Real-Time PCR(RT-PCR)测定了MnCP-3基因在头胸甲中蜕皮间期(C)、蜕皮前早期(D0-2)、蜕皮前晚期(D3-4)和蜕皮后期(A)4个不同蜕皮时期中的表达水平以及KK-42处理后表达水平的变化.结果显示,MnCP-3在D3-4期和A期表达较高.KK-42处理后,C期MnCP-3的表达水平在处理后3～48h相对于对照组显著升高,在D0-2期MnCP-3的表达水平在3、6、24和48h显著升高.结果提示,MnCP-3可能参与新的外表皮和内表皮形成,KK-42处理可显著诱导C期和D0-2期MnCP-3的表达,提前启动蜕皮过程的进行,加快新表皮的形成.

In order to analyze the effect of imidazole derivative KK-42 on expressions of Macrobrachium nipponense cuticular proteins （MnCPs） gene preliminarily, a novel cuticular protein gene named Macrobrachium nipponense cuticular protein-3 （ MnCP-3 ） containing chitin_bind_4 conserve domain was cloned from carapace. The relative expression of MnCP-3 in carapace during four molt stages-intermolt stage （C）, early premolt stage （D 0-2 ）, late premolt stage （D 3-4 ） and postmolt stage （A） and the effect after KK-42 treatment were measured by Real-time PCR （RT-PCR）. The results showed that the relative expression of MnCP-3 was higher during stages D 3-4 and A. What’s more, the expression levels were higher than that of control from 3 to 48 h during stage C and at 3, 6, 24 and 48 h during D 0-2 after KK-42 treatment. It suggests that MnCP-3 is likely involved in forming of new exocuticle and endocuticle, and the expression of MnCP-3 could be induced significantly at stage C and D 0-2 after KK-42 treatment, which results in starting molting process in advance and accelerating formation of new cuticle.