P-001 ClpX is required in maintaining mitochondrial functions during meiosis and spermatogenesis

Abstract Study question What is the role of ClpX in spermatogonia differentiation and spermatogenesis? Summary answer required to maintain normal mitochondrial functions spermatocytes, deficiency resulted meiosisarrest diruption spermatogenesis mice. known already mitochondria-specific quality control protease, it maintains proteostasis via degrading misfolded or damagedproteins matrix. a AAA+ protease that uses energy ATP binding hydrolysis degradeunfolded proteins. reported working as complex with ClpP, consists hexamer ofClpX tetradecamer ClpP. recognizes protein substrates their unstructured peptidesequences, termed degradation tags recognition signals. The fragments cleaved polypeptides can then exitthe chamber be further degraded. design, size, duration design: we generated germ cell specific knockout mice line evaluated weight, inner structure oftestis tissue. We also applied biochemical techniques high-throughput sequencing investigate themechanism regulating spermatogenesis. Size: more than 50 were used this study, including Duration: 2 years Participants/materials, setting, methods Participants/materials: conditional (cKO) by CRISPR-gene editing Cre-LoxP system, which specifically cells male Setting: siblings WT cKO compared various experiments. Mehods: performed morphology comparement check weight size tetes cKOmice, immunofluorescence, western blot, histological andpharmocological treatment. Main results chance found reduced quantity affected supply duringmeiosis attenuated zygotene-pachytene transformation cells. spermatocytesexhibited disorder chromosome synapsis cross-over events. Their telomeres failed attach nuclearenvelop, was associated failure α-tubulin formation deficient spermatocytes. dysregulatedspermatocytes finally underwent apoptosis resulting decreased testicular vacuolar structures within theseminiferous tubules. Both transcriptome analysis m6A-methylation highlighted dysregulationof metabolic pathways mTOR signaling isolated confirmed over-activation themTORC1 pathway increased expression phosphorylated S6 4EBP after deletion spermatocytes.Long-term inhibition mTORC1 rapamycin treatment vivo partially rescue tubules much less apoptotic late stage meiotic cells, such asround spermatids elongated spermatozoon. data reveal novel roles meiosis andspermatogenesis. Limitations, reasons for caution This study based on animal experiments, thus, these only indicated important micespermatogenesis rather human beings. Clinical has not yet Clpx gene mutation reproductivedisease. Wider implications findings report ClpP induce autosomal recessive Perrault syndrome, while themutation been from clinical data. Our resutls demonstrated severe effect onspermatogenesis knockingout ClpX, might give some insights clinic. Trial registration number applicable