Analysis Of The Expression Patterns Of MIH, EcR And RXR Genes In Procambarus Clarkii During The Molting Process And Preliminary Exploration Of Their Interaction

The full-length cDNA of PcMIH,PcEcR and PcRXR from the crayfish Procambarus clarkii were cloned in this study.The gene sequence,functional structure,physical and chemical properties analyses,etc of PcMIH,PcEcR and PcRXR were performed by using bioinformatical approaches.The Real-time quantitative PCR(RT-qPCR)was used to detect the relative expression level of the three genes in different tissues and molt stages.The results indicated that the PcMIH was detected only in eyestalk.The PcEcR and PcRXR were detected in all tissues.Both of them showed the highest level in gill and hepatopancreas and the two groups of expression showed similar results.The expression level of PcMIH in eyestalk increased from postmolt,reached a maximum in intermolt and then decreased in premolt.The expression level of PcEcR increased from postmolt and reached a maximum in early premolt and then decreased in eyestalk,hepatopancreas and muscle.But in gill,the expression level of PcEcR increased from postmolt and reached a maximum in intermolt and then decreased in whole premolt.The expression level of PcRXR showed a same fluctuating trends in eyestalk,hepatopancreas,muscle and gill which decreased from postmolt to late premolt.Both unilateral and bilateral eyestalk ablation resulted in the increasing expression level of PcEcR in hepatopancreas and gill in three days and falled at the 7th day.A same result of PcRXR in gill suggested that PcMIH can inhibit the expression of PcEcR and PcRXR.On the other hand,the expression level of PcRXR kept increasing after both unilateral and bilateral eyestalk ablation in hepatopancreas in seven days.A possible reason is eyestalk ablation can promote the produce of vitellogenin(Vg)in hepatopancreas and PcRXR is one of the genes associated with the produce of Vg.As a result of RNA interference,different tissue showed different sensitivity of RNAi.An obvious decreasing expression level of PcEcR was found after RNAi suggesting a collaborative relationship between PcEcR and PcRXR.The expression level of PcMIH also decreased obviously,the reason not clear,and more experiments were needed.