Genetic Mutations And Function Detections In Autosomal Dominant Optic Atrophy

Purpose: Autosomal dominant optic atrophy(ADOA),is a primary degeneration of the ganglion cell layer in the retina,accompanied with ascending optic atrophy.It is a common optic atrophy which can lead moderate to severe visual impairment.The study aims to collect the blood sample from families with ADOA and use the whole exome sequencing(WES)to detect,sanger sequencing to evaluate the mutation sites.Then explore the mutate site and evaluate the structure and function of mutations.Methods: The blood of the family with ADOA was gathered to take a genome DNA sample.Using WES to detect,sanger sequencing to evaluate the mutation sites.PyMol and Schr?dinger are used to construct the mutate protein structure and detect the function in molecular dynamics simulation.Three kinds of plasmids,including wild type(WT),negative control(NC)and mutant type(MT),were built to transduce to RGC 5.The function of mutations was evaluated in the content of cleaved-caspase-3 of each group by western blot and mitochondrial network morphology analysis.Results: In the family,the mutation(NM＿130837:exon10 c.1034G>A:p.R345Q)was assayed.The mutate site is located in GTPase domain.The stability of mutate protein is decreased,and the protein-ligand contact decreased by using molecular dynamics simulation.Compared to the WT and NC,the expression of cleaved-caspase-3 in MT are increased significantly(p<0.001).Compared to the NC,the counts of individuals increased(p=0.014),the mean branch length decreased(p<0.001),and the counts of individuals/ the counts of networks increased in MT(p<0.001).The mitochondrial network morphology changed a lot in Mutant type.Conclusion: The study proved that the stability of mutate protein is decreased,and the protein-ligand contact decreased by using molecular dynamics simulation.Besides,the OPA1 mutation could damage the mitochondrial network morphology,increase the expression of cleaved-caspase-3 and may be relevant with apoptosis.