title: Rhodopsin mutant P23H destabilizes rod photoreceptor disk membranes.
abstract: Mutations in rhodopsin cause retinitis pigmentosa in humans and retinal degeneration in a multitude of other animals. We utilized high-resolution live imaging of the large rod photoreceptors from transgenic frogs (Xenopus) to compare the properties of fluorescently tagged rhodopsin, Rho-EGFP, and Rho(P23H)-EGFP. The mutant was abnormally distributed both in the inner and outer segments (OS), accumulating in the OS to a concentration of ∼0.1% compared to endogenous opsin. Rho(P23H)-EGFP formed dense fluorescent foci, with concentrations of mutant protein up to ten times higher than other regions. Wild-type transgenic Rho-EGFP did not concentrate in OS foci when co-expressed in the same rod with Rho(P23H)-EGFP. Outer segment regions containing fluorescent foci were refractory to fluorescence recovery after photobleaching, while foci in the inner segment exhibited recovery kinetics similar to OS regions without foci and Rho-EGFP. The Rho(P23H)-EGFP foci were often in older, more distal OS disks. Electron micrographs of OS revealed abnormal disk membranes, with the regular disk bilayers broken into vesiculotubular structures. Furthermore, we observed similar OS disturbances in transgenic mice expressing Rho(P23H), suggesting such structures are a general consequence of mutant expression. Together these results show that mutant opsin disrupts OS disks, destabilizing the outer segment possibly via the formation of aggregates. This may render rods susceptible to mechanical injury or compromise OS function, contributing to photoreceptor loss.
authors.affiliation: Center for Vision Research, Department of Neuroscience, State University of New York Upstate Medical University, Syracuse, New York, United States of America.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Haeri
authors.forename: Barry E
authors.initials: BE
authors.lastname: Knox
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D007150:Immunohistochemistry; D007814:Larva / Q000378:metabolism; D018613:Microscopy, Confocal; D009154:Mutation; D012160:Retina / Q000378:metabolism; D017948:Retinal Rod Photoreceptor Cells / Q000378:metabolism; D012243:Rhodopsin / Q000235:genetics / Q000378:metabolism*; D014981:Xenopus
chemical_list: D012243:Rhodopsin