CEP290 localization in the rod connecting cilium of CEP290rd16 mice with fluorescence nanoscopy

摘要: Purpose: Mutations in CEP290 account for roughly 25% of cases of Leber Congenital Amaurosis (LCA). It is associated with the photoreceptor connecting cilia (CC) but its function is not certain. We used an LCA-like mouse model, homozygous for the CEP290 rd16 allele, with an in-frame deletion of the putative microtubule binding domain, to assess the function of this domain. We hypothesized that CEP290 and the CEP290 interacting partner, NPHP5, mislocalize in CEP290 rd16 photoreceptors and that this mislocalization contributes to the rapid retinal degeneration in the CEP290 rd16 animal.Methods: To obtain sub-diffraction resolution images, we used Structured Illumination Microscopy (SIM) in age-matched wild-type and mutant mice. In order to localize CEP290 prior to retinal degeneration, retinas from 10-day old mice were immunostained with antibodies for CEP290 and antibodies to other CC proteins …