Rhodopsin transport in the membrane of the connecting cilium of mammalian photoreceptor cells.
The transport of the photopigment rhodopsin from the inner segment to the photosensitive outer segment of vertebrate photoreceptor cells has been one of the main remaining mysteries in photoreceptor cell biology. Because of the lack of any direct evidence for the pathway through the photoreceptor cilium, alternative extracellular pathways have been proposed. Our primary aim in the present study was to resolve rhodopsin trafficking from the inner to the outer segment. We demonstrate, predominantly by high-sensitive immunoelectron microscopy, that rhodopsin is also densely packed in the membrane of the photoreceptor connecting cilium. Present prominent labeling of rhodopsin in the ciliary membrane provides the first striking evidence that rhodopsin is translocated from the inner segment to the outer segment of wild type photoreceptors via the ciliary membrane. At the ciliary membrane rhodopsin co-localizes with the unconventional myosin VIIa, the product of human Usher syndrome 1B gene. Furthermore, axonemal actin was identified in the photoreceptor cilium, which is spatially co-localized with myosin VIIa and opsin. This actin cytoskeleton of the cilium may provide the structural bases for myosin VIIa-linked ciliary trafficking of membrane components, including rhodopsin.
Wolfrum
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Institute of Zoology, Johannes Gutenberg-University of Mainz, Mainz, Germany. wolfrum@mail.uni-mainz.de
Schmitt
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D000199:Actins / Q000032:analysis / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001692:Biological Transport; D002417:Cattle; D002923:Cilia / Q000737:chemistry / Q000378:metabolism; D004398:Dyneins; D005260:Female; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D016253:Microscopy, Immunoelectron; D008875:Middle Aged; D008969:Molecular Sequence Data; D000081412:Myosin VIIa; D009218:Myosins / Q000032:analysis / Q000276:immunology; D051381:Rats; D017207:Rats, Sprague-Dawley; D017948:Retinal Rod Photoreceptor Cells / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D012174:Retinitis Pigmentosa / Q000235:genetics; D012243:Rhodopsin / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000378:metabolism* / Q000648:ultrastructure*; D017299:Rod Opsins / Q000032:analysis / Q000276:immunology
D000199:Actins; D000911:Antibodies, Monoclonal; C000631455:MYO7A protein, human; C000631456:Myo7a protein, mouse; C000631457:Myo7a protein, rat; D000081412:Myosin VIIa; D017299:Rod Opsins; D012243:Rhodopsin; D009218:Myosins; D004398:Dyneins