The Effect Of Fam73a Gene On Auditory Function

Objective:to study the effect of fam73a gene on auditory functionMethods:1,A fam73a gene knockout mouse model was constructed by TALEN technology.2,RT-PCR and immunohistochemistry were used to observe the spatiotemporal expression of fam73a in the inner ear.3,audiometry changes of fam73a gene knockout mice through ABR detection.Scanning electron microscopy was performed to observe the morphological changes of cochlear hair cells in fam73a gene knockout mice.Results:First of all,we established a mouse model of the fam73a gene knockout using the TALEN technique:This mouse model was derived from the laboratory of Prof.jinjin.And by sequencing,PCR,Western Blot and other experimental methods to verify the gene knockout success.To investigate the effect of fam73a gene deletion on hearing,4 fam73a knockout mice of P30 and 3 wild type mice of the same age were used.ABR was measured after anesthesia with pentobarbital sodium,and the results were analyzed.After knocking out of fam73a gene,it causes hearing loss in mice.Anatomical basement membranes were immunofluorescently stained with hair cell marker myosin7a.We found that there was a punctate deletion of hair cells in the basement membrane at the base of the basement membrane.Based on the statistical analysis of this missing phenomenon,it was found that at the time of P30,hair cells of the inner ear hair cells of fam73a knockout mice were missing in the bottom circle.In order to continue to study the effect of fam73a gene deletion on hearing,5 fam73a knockout mice of P60 and 4 wild type mice of the same age were used to measure ABR after anesthesia with pentobarbital sodium,and the results were analyzed.It was found that fam73a knockouts cause severe hearing loss in mice.Anatomical basement membranes were immunofluorescently stained with the hair cell marker myosin7a.We found that the basement membranes had punctate deletions of hair cells from the top circle to the bottom circle.The absence of this type of hair cells was the outer hair cell,and a statistical analysis was performed on this missing phenomenon.It was found that the loss of inner ear hair cells from fam73a knockout mice increased from the top circle to the bottom circle at P60.Conclusion:after fam73a gene knockout Adult mouse hearing loss,...