Immunogold localization of actin in developing photoreceptor cilia of normal and rds mutant mice

Abstract

Recent immunocytochemical studies have localized actin to the distal cilium of mature vertebrate photoreceptors. Since this is the site where the ciliary plasma membrane evaginates to form new outer segment discs, the results suggest that an actin-mediated contractile mechanism or cytoskeletal network might regulate some aspect of outer segment disc morphogenesis. In the present study, immunoelectron microscopy was used to localize actin to the developing cilia of normal and rds mutant mouse photoreceptors. In normal mice, actin could not be localized to newly projecting cilia, but an actin-rich domain was demonstrated within the distal, bulbous ending of elongated cilia just prior to outer segment development. These results suggest that actin is not important for ciliary growth, but that it may be necessary for the subsequent differentiation of an outer segment. In the rds mutant mouse, there is an absence of outer segment formation, although cilia appear to develop normally. Rhodamine phalloidin staining of cryostat sections demonstrated a normal F-actin distribution within the rds retina. Utilizing immunogold labeling of developing rds photoreceptors, actin was localized to the distal, bulbous ending of elongated cilia. This result indicates that actin is situated within its normal domain in rds cilia.