The Effects Of Regulation Of AMPK On PD Transgenic Drosophila Model

Objective: In order to study whether there exists the neuroprotective role of regulated expression of AMPK in the PINK1 mutant related PD transgenic Drosophila model and the related mechanism. Methods : Utilizing the classical PD transgenic Drosophila model—PINK1^B9,which is dominantly modified PD related gene,PINK1,null mutant. Here, using the classical model of GAL/UAS system, and the MHC-GAL4 promotor to make the disease characteristics specifically expressed in Drosophila muscles, thus construct the MHC-Gal4 system of PD transgenic Drosophila model. Using Western Blot to measure the expression of activated form of AMPK to investigate whether the activation of AMPK is abnormal in the mutant form of PD Drosophila model. Furthermore, using MHC-Gal4 to drive AMPK over expressed or down regulated by RNA interference in PD transgenic Drosophila model to study the role of AMPK in the pathogenisis of PD. Observe the abnormal wing posture and flying rate of PD transgenic flies and the ATP level in the muscles and measure the ND42 mRNA level in mitochondrial respiratory chain oxide compound I and the expression of the corresponding protein—NDUFS-3 level to evaluate the alteration of the mitochondrial function. Study the role of regulated AMPK in the pathogenesis of PD and the related mechanism. Results: We found that the main activated form of AMPK—P-AMPK-α decreased in the mutant form of PD Drosophila model compared with the normal controls. No matter using the over expressed or down regulated form of AMPK to interfere PINK^B9 were able to rescue the abnormal wing posture and flying rate of PD transgenic flies, and both of the regulation could correspondingly alter the expression of P-AMPK-α.When detecting the mitochondrial function in the AMPK regulated PD transgenic flies, we found that the severe mitochondrial dysfunction in PD flies has been significantly ameliorated, and the ATP level notably increased, the morphology of mitochondria in muscle fibers has been improved with the improvement of ATP level, the amount of ND42 mRNA and the expression of NDUFS3 significantly increased, both of which mark the function of mitochondrial oxidative respiratory chain Complex I. Conclusions: Not only up regulation of AMPK could rescue the PD flies but also down regulated form could directly improve mitochondrial function of PINK1^B9 Drosophila model.