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      The route of the visual receptor rhodopsin along the cilium

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          ABSTRACT

          The photoreceptor outer segment is the most elaborate primary cilium, containing large amounts of rhodopsin (RHO) in disk membranes that grow from a connecting cilium. The movement of RHO along the connecting cilium precedes formation of the disk membranes. However, the route that RHO takes has not been clearly determined; some reports suggest that it follows an intracellular, vesicular route along the axoneme, possibly as an adaptation for the high load of delivery or the morphogenesis of the disk endomembranes. We addressed this question by studying RHO in cilia of IMCD3 cells and mouse rod photoreceptors. In IMCD3 cilia, fluorescence recovery after photobleaching (FRAP) experiments with fluorescently tagged RHO supported the idea of RHO motility in the ciliary plasma membrane and was inconsistent with the hypothesis of RHO motility within the lumen of the cilium. In rod photoreceptors, FRAP of RHO–EGFP was altered by externally applied lectin, supporting the idea of plasmalemmal RHO dynamics. Quantitative immunoelectron microscopy corroborated our live-cell conclusions, as RHO was found to be distributed along the plasma membrane of the connecting cilium, with negligible labeling within the axoneme. Taken together, the present findings demonstrate RHO trafficking entirely via the ciliary plasma membrane.

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          Abstract

          [Related article:] Highlighted Article: The unresolved question of the route that rhodopsin travels along the cilium to accommodate disk membrane morphogenesis was addressed; it was found to be exclusively plasmalemmal.

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          Author and article information

          Journal
          J Cell Sci
          J. Cell. Sci
          JCS
          joces
          Journal of Cell Science
          The Company of Biologists Ltd
          0021-9533
          1477-9137
          15 May 2019
          15 May 2019
          15 May 2020
          : 132
          : 10
          : jcs229526
          Affiliations
          [1 ] Departments of Ophthalmology and Neurobiology, Jules Stein Eye Institute, David Geffen School of Medicine at UCLA, University of California , Los Angeles, CA 90095, USA
          [2 ] Molecular Biology Institute, University of California , Los Angeles, CA 90095, USA
          [3 ] Brain Research Institute, University of California , Los Angeles, CA 90095, USA
          Author notes
          [* ]Author for correspondence ( dswilliams@ 123456ucla.edu )
          Author information
          http://orcid.org/0000-0002-7758-3932
          Article
          PMC6550008 PMC6550008 6550008 JCS229526
          10.1242/jcs.229526
          6550008
          30975916
          d489ae46-bdc9-4826-80e8-e1470bbf5e70
          © 2019. Published by The Company of Biologists Ltd
          History
          : 3 January 2019
          : 2 April 2019
          Funding
          Funded by: National Institutes of Health, http://dx.doi.org/10.13039/100000002;
          Award ID: F32EY026318
          Award ID: R01EY013408
          Award ID: EY027442
          Award ID: P30EY0331
          Categories
          136
          Research Article

          Photoreceptor,Rhodopsin,Trafficking,Cilium
          Photoreceptor, Rhodopsin, Trafficking, Cilium

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