Ciliary Transport of Opsin

  • Deepti Trivedi
  • David S. Williams
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 664)


As part of the renewal of photoreceptor outer segment disk membranes, membrane proteins are transported along the region of the cilium, connecting the inner and outer segments. Genetics studies have indicated the role of motor proteins in this transport. Direct analysis of live cells is needed to increase our understanding of the transport mechanisms further. Here, we show that transfection of hTERT-RPE1 cells with constructs encoding RHO-EGFP, but not RHO-mCherry, results in the distribution of fluorescently-tagged opsin in the plasma membrane. When the cells have differentiated and possess cilia, a portion of the RHO-EGFP was observed along the cilia. Due to the remarkable conservation of ciliary protein function, this system of Rho-Egfp transfected hTERT-RPE1 cells provides a valid model with which to study the ciliary transport of opsin directly in live cells.


Outer Segment Disk Membrane Photoreceptor Outer Segment Joubert Syndrome Intraflagellar Transport 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We thank Dr Zhaohuai Yang, UCSD, for providing with the Rho-Gfp and Rho-mCherry constructs, and Dr Carolyn Ott, NIH, for providing with the Smo-CFP construct. We also thank Dr Vanda Lopes for providing the hTERT-RPE1 cell line.


  1. Beech PL, Pagh Roehl K, Noda Y et al (1996) Localization of kinesin superfamily proteins to the connecting cilium of fish photoreceptors. J Cell Sci 109:889–897PubMedGoogle Scholar
  2. Besharse JC, Forestner DM, Defoe DM (1985) Membrane assembly in retinal photoreceptors. III. Distinct membrane domains of the connecting cilium of developing rods. J Neurosci 5:1035–1048PubMedGoogle Scholar
  3. Besharse JC, Wetzel MG (1995) Immunocytochemical localization of opsin in rod photoreceptors during periods of rapid disc assembly. J Neurocytol 24:371–388CrossRefPubMedGoogle Scholar
  4. Blacque OE, Leroux MR (2006) Bardet-Biedl syndrome: an emerging pathomechanism of intracellular transport. Cell Mol Life Sci 63:2145–2161CrossRefPubMedGoogle Scholar
  5. Brown SS (1999) Cooperation between microtubule- and actin-based motor proteins. Ann Rev Cell Dev Biol 8:1751–1755Google Scholar
  6. Chuang JZ, Sung CH (1998) The cytoplasmic tail of rhodopsin acts as a novel apical sorting signal in polarized MDCK cells. J Cell Biol 142:1245–1256CrossRefPubMedGoogle Scholar
  7. Cole DG, Diener DR, Himelblau AL et al (1998) Chlamydomonas kinesin-II-dependent intraflagellar transport (IFT): IFT particles contain proteins required for ciliary assembly in Caenorhabditis elegans sensory neurons. J Cell Biol 141:993–1008CrossRefPubMedGoogle Scholar
  8. Fliegauf M, Benzing T, Omran H (2007) When cilia go bad: cilia defects and ciliopathies. Nat Rev Mol Cell Biol 8:880–893CrossRefPubMedGoogle Scholar
  9. Insinna C, Pathak N, Perkins B et al (2008) The homodimeric kinesin, Kif17, is essential for vertebrate photoreceptor sensory outer segment development. Dev Biol 316:160–170CrossRefPubMedGoogle Scholar
  10. Jimeno D, Feiner L, Lillo C et al (2006) Analysis of kinesin-2 function in photoreceptor cells using synchronous Cre-loxP knockout of Kif3a with RHO-Cre. Invest Ophthalmol Vis Sci 47:5039–5046CrossRefPubMedGoogle Scholar
  11. Jin S, McKee TD, Oprian DD (2003) An improved rhodopsin/EGFP fusion protein for use in the generation of transgenic Xenopus laevis. FEBS Lett 542:142–146CrossRefPubMedGoogle Scholar
  12. Kozminski KG, Beech PL, Rosenbaum JL (1995) The Chlamydomonas kinesin-like protein FLA10 is involved in motility associated with the flagellar membrane. J Cell Biol 131:1517–1527CrossRefPubMedGoogle Scholar
  13. Liu X, Udovichenko IP, Brown SDM et al (1999) Myosin VIIa participates in opsin transport through the photoreceptor cilium. J. Neurosci 19:6267–6274PubMedGoogle Scholar
  14. Marszalek JR, Liu X, Roberts EA et al (2000) Genetic evidence for selective transport of opsin and arrestin by kinesin-II in mammalian photoreceptors. Cell 102:175–187CrossRefPubMedGoogle Scholar
  15. Moritz OL, Tam BM, Papermaster DS et al (2001) A functional rhodopsin-green fluorescent protein fusion protein localizes correctly in transgenic Xenopus laevis retinal rods and is expressed in a time-dependent pattern. J Biol Chem 276:28242–28251CrossRefPubMedGoogle Scholar
  16. Nir I, Papermaster DS (1983) Differential distribution of opsin in the plasma membrane of frog photoreceptors: an immunocytochemical study. Invest Ophthalmol Vis Sci 24:868–878PubMedGoogle Scholar
  17. Orozco JT, Wedaman KP, Signor D et al (1999) Movement of motor and cargo along cilia [letter]. Nature 398:674CrossRefPubMedGoogle Scholar
  18. Otto EA, Loeys B, Khanna H et al (2005) Nephrocystin-5, a ciliary IQ domain protein, is mutated in Senior-Loken syndrome and interacts with RPGR and calmodulin. Nat Genet 37:282–288CrossRefPubMedGoogle Scholar
  19. Pazour GJ, Baker SA, Deane JA et al (2002) The intraflagellar transport protein, IFT88, is essential for vertebrate photoreceptor assembly and maintenance. J Cell Biol 157:103–113CrossRefPubMedGoogle Scholar
  20. Richardson TM (1969) Cytoplasmic and ciliary connections between the inner and outer segments of mammalian visual receptors. Vision Res 9:727–731CrossRefPubMedGoogle Scholar
  21. Rosenbaum JL, Witman GB (2002) Intraflagellar transport. Nat Rev Mol Cell Biol 3:813–825CrossRefPubMedGoogle Scholar
  22. Sayer JA, Otto EA, O‘Toole JF et al (2006) The centrosomal protein nephrocystin-6 is mutated in Joubert syndrome and activates transcription factor ATF4. Nat Genet 38:674–681CrossRefPubMedGoogle Scholar
  23. Taipale J, Chen JK, Cooper MK et al (2000) Effects of oncogenic mutations in Smoothened and Patched can be reversed by cyclopamine. Nature 406:1005–1009CrossRefPubMedGoogle Scholar
  24. Williams DS (2002) Transport to the photoreceptor outer segment by myosin VIIa and kinesin II. Vision Res 42:455–462CrossRefPubMedGoogle Scholar
  25. Wolfrum U, Schmitt A (2000) Rhodopsin transport in the membrane of the connecting cilium of mammalian photoreceptor cells. Cell Motil Cytoskeleton 46:95–107CrossRefPubMedGoogle Scholar
  26. Young RW (1967) The renewal of photoreceptor cell outer segments. J. Cell Biol 33:61–72CrossRefPubMedGoogle Scholar
  27. Young RW (1968) Passage of newly formed protein through the connecting cilium of retina rods in the frog. J Ultrastruct Res 23:462–473CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Jules Stein Eye Institute, University of CaliforniaLos AngelesUSA

Personalised recommendations