The Effects Of Sidt2 Mecganism On Skeletel Muscle Autophag And Mitochondrial Encephalopathy Due To Mutation In NDUFAF7

Objective Study the effects of Sidt2 mechanism on Skeletal Muscle Autophagic SignalingMethods(1)Generate the skeletal muscle-conditional Sidt2 knockout mice and control mice,using RT-PCR and Western Blot analysis Sidt2 m RNA and protein level;(2)Compare the mass of gastrocnemius、tibialis anterior、and the soleus weight with the control mice;(3)Compare Muscle strength level of two groups of mice;(4)ATPase staining analysis the content of fast-twitch muscle fibers and slow-twitch muscle fibers in muscle group;(5)The activity of CK and the content ATP 、collagen were analyzed by kits.(6)The expression level of autophagy related proteins m TORC1、S6K1 were detected by Western Blot;(7)Combined plasmid transfection with IP technology to find protein interacted with Sidt2.Results Both m RNA and protein level of Sidt2 were significantly lower in musclespecific Sidt2 knockout mice compared with control mice;The absolute weight and relative weight of GC and TA were decrease in muscle-specific Sidt2 knockout mice compared with control mice,but not for SOL muscles;The level of muscle strength level was significantly decreased in muscle-specific Sidt2 knockout mice;ATPase staining suggested that the content of fast-twitch muscle fibers and slow-twitch muscle fibers was changed in model muscle;The activity of CK and the content of collagen were increased,and the content of ATP was decreased;The expression level of p-m TORC1、p-S6K1 was increased showed by Western Blot;By using IP technology,NPC1 protein was precipitation down in overexpress Sidt2 cells.Conclusion Mice with muscle-specific deletion of the lysosomal membrane gene Sidt2 showed symptoms of primary muscular dystrophy-like phenotype.Sidt2 can affect m TORC1 pathway and may interact with NPC1 protein.Objective This study reviews a case of mitochondrial respiratory chain complex I deficiency due to the double mutation in mitochondrial NDUFAF7 gene.Methods The patient came from Hospital,Shanghai Jiao Tong University School.The patient presented with mental retardation,motor dysfunction,epilepsy.(MRI),blood lactic acid was normal;WES found NDUFAF7 double allele variants: c.355 c > T(p.G ln119 *)and c.842 T > c(p.I le281Thr).To confirm the diagnosis,construction of patient lymphocyte line patients and analysis the NUDFAF7 protein,complex enzyme activity level and BN-PAGE I detection.Results The results showed m RNA and protein level of NDUFAF7 was decrease.Mitochondrial function was impaired;Activity of complex I was lower than control cells;BN-PAGE showed complex I deficiency.Conclusion It is concluded that the patient diagnosed as complex I deficiency due to c.355 c > T(p.G ln119 *)and c.842 T > c(p.I le281Thr)mutation in NDUFAF7.