Mutations in Pre-mRNA Processing Factors 3, 8, and 31 Cause Dysfunction of the Retinal Pigment Epithelium

• DOI: 10.1016/j.ajpath.2014.06.026
• Publication Date: 2014-08-08T13:00:28Z
• Authors: Michael H. Farkas; Deborah S. Lew; Maria E. Sousa; Kinga Bujakowska; Jonathan Chatagnon; Shomi S. Bhattach...; Eric A. Pierce; Emeline F. Nandrot
• Keywords: Male; 0301 basic medicine; RNA Splicing; Mice, Transgenic; Retinal Pigment Epithelium; Retina; Pathology and Forensic Medicine; Mice; 03 medical and health sciences; Phagocytosis; RNA Precursors; Animals; Humans; [SDV.MHEP.OS]Life Sciences [q-bio]/Human health and pathology/Sensory Organs; Eye Proteins; Retinal Degeneration; RNA-Binding Proteins; Circadian Rhythm; Disease Models, Animal; [SDV.MHEP.OS] Life Sciences [q-bio]/Human health and pathology/Sensory Organs; Gene Knockdown Techniques; Mutation; Female; RNA Splicing Factors; Retinitis Pigmentosa

Mutations in the ubiquitously expressed pre-mRNA processing factors 3, 8, and 31 (PRPF3, PRPF8, and PRPF31) cause nonsyndromic dominant retinitis pigmentosa in humans, an inherited retinal degeneration. It is unclear what mechanisms, or which cell types of the retina, are affected. Transgenic mice with the human mutations in these genes display late-onset morphological changes in the retinal pigment epithelium (RPE). To determine whether the observed morphological changes are preceded by abnormal RPE function, we investigated its phagocytic function in Prpf3(T494M/T494M), Prpf8(H2309P/H2309P), and Prpf31(+/-) mice. We observe decreased phagocytosis in primary RPE cultures from mutant mice, and this is replicated by shRNA-mediated knockdown of PRPF31 in human ARPE-19 cells. The diurnal rhythmicity of phagocytosis is almost lost, indicated by the marked attenuation of the phagocytic burst 2 hours after light onset. The strength of adhesion between RPE apical microvilli and photoreceptor outer segments also declined during peak adhesion in all mutants. In all models, at least one of the receptors involved in binding and internalization of shed photoreceptor outer segments was subjected to changes in localization. Although the mechanism underlying these changes in RPE function is yet to be elucidated, these data are consistent with the mouse RPE being the primary cell affected by mutations in the RNA splicing factors, and these changes occur at an early age.