Search tips
Search criteria 


Logo of jcellbiolHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
J Cell Biol. 1991 June 2; 113(6): 1281–1293.
PMCID: PMC2289036

Polarized sorting of rhodopsin on post-Golgi membranes in frog retinal photoreceptor cells


We have isolated a subcellular fraction of small vesicles (mean diameter, 300 nm) from frog photoreceptors, that accumulate newly synthesized rhodopsin with kinetics paralleling its appearance in post- Golgi membranes in vivo. This fraction is separated from other subcellular organelles including Golgi and plasma membranes and synaptic vesicles that are sorted to the opposite end of the photoreceptor cell. The vesicles have very low buoyant density in sucrose gradients (rho = 1.09 g/ml), a relatively simple protein content and an orientation of rhodopsin expected of transport membranes. Reversible inhibition of transport by brefeldin A provides evidence that these vesicles are exocytic carriers. Specific immunoadsorption bound vesicles whose protein composition was indistinguishable from the membranes sedimented from the subcellular fraction. Some of these proteins may be cotransported with rhodopsin to the rod outer segment; others may be involved in vectorial transport.

Full Text

The Full Text of this article is available as a PDF (3.8M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Balch WE, Dunphy WG, Braell WA, Rothman JE. Reconstitution of the transport of protein between successive compartments of the Golgi measured by the coupled incorporation of N-acetylglucosamine. Cell. 1984 Dec;39(2 Pt 1):405–416. [PubMed]
  • Bartles JR, Feracci HM, Stieger B, Hubbard AL. Biogenesis of the rat hepatocyte plasma membrane in vivo: comparison of the pathways taken by apical and basolateral proteins using subcellular fractionation. J Cell Biol. 1987 Sep;105(3):1241–1251. [PMC free article] [PubMed]
  • Bennett MK, Wandinger-Ness A, Simons K. Release of putative exocytic transport vesicles from perforated MDCK cells. EMBO J. 1988 Dec 20;7(13):4075–4085. [PubMed]
  • Berryman MA, Rodewald RD. An enhanced method for post-embedding immunocytochemical staining which preserves cell membranes. J Histochem Cytochem. 1990 Feb;38(2):159–170. [PubMed]
  • Besharse JC, Pfenninger KH. Membrane assembly in retinal photoreceptors I. Freeze-fracture analysis of cytoplasmic vesicles in relationship to disc assembly. J Cell Biol. 1980 Nov;87(2 Pt 1):451–463. [PMC free article] [PubMed]
  • Burger PM, Mehl E, Cameron PL, Maycox PR, Baumert M, Lottspeich F, De Camilli P, Jahn R. Synaptic vesicles immunoisolated from rat cerebral cortex contain high levels of glutamate. Neuron. 1989 Dec;3(6):715–720. [PubMed]
  • Chavrier P, Parton RG, Hauri HP, Simons K, Zerial M. Localization of low molecular weight GTP binding proteins to exocytic and endocytic compartments. Cell. 1990 Jul 27;62(2):317–329. [PubMed]
  • Cooper MS, Cornell-Bell AH, Chernjavsky A, Dani JW, Smith SJ. Tubulovesicular processes emerge from trans-Golgi cisternae, extend along microtubules, and interlink adjacent trans-golgi elements into a reticulum. Cell. 1990 Apr 6;61(1):135–145. [PubMed]
  • De Camilli P, Harris SM, Jr, Huttner WB, Greengard P. Synapsin I (Protein I), a nerve terminal-specific phosphoprotein. II. Its specific association with synaptic vesicles demonstrated by immunocytochemistry in agarose-embedded synaptosomes. J Cell Biol. 1983 May;96(5):1355–1373. [PMC free article] [PubMed]
  • de Curtis I, Simons K. Isolation of exocytic carrier vesicles from BHK cells. Cell. 1989 Aug 25;58(4):719–727. [PubMed]
  • Defoe DM, Besharse JC. Membrane assembly in retinal photoreceptors. II. Immunocytochemical analysis of freeze-fractured rod photoreceptor membranes using anti-opsin antibodies. J Neurosci. 1985 Apr;5(4):1023–1034. [PubMed]
  • Deretic D, Hamm HE. Topographic analysis of antigenic determinants recognized by monoclonal antibodies to the photoreceptor guanyl nucleotide-binding protein, transducin. J Biol Chem. 1987 Aug 5;262(22):10839–10847. [PubMed]
  • Dunn WA, Hubbard AL. Receptor-mediated endocytosis of epidermal growth factor by hepatocytes in the perfused rat liver: ligand and receptor dynamics. J Cell Biol. 1984 Jun;98(6):2148–2159. [PMC free article] [PubMed]
  • Ey PL, Prowse SJ, Jenkin CR. Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A-sepharose. Immunochemistry. 1978 Jul;15(7):429–436. [PubMed]
  • Fanger BO. Adaptation of the Bradford protein assay to membrane-bound proteins by solubilizing in glucopyranoside detergents. Anal Biochem. 1987 Apr;162(1):11–17. [PubMed]
  • Fukuda MN, Papermaster DS, Hargrave PA. Rhodopsin carbohydrate. Structure of small oligosaccharides attached at two sites near the NH2 terminus. J Biol Chem. 1979 Sep 10;254(17):8201–8207. [PubMed]
  • Greenberger LM, Besharse JC. Photoreceptor disc shedding in eye cups. Inhibition by deletion of extracellular divalent cations. Invest Ophthalmol Vis Sci. 1983 Nov;24(11):1456–1464. [PubMed]
  • Hall MO, Bok D, Bacharach AD. Biosynthesis and assembly of the rod outer segment membrane system. Formation and fate of visual pigment in the frog retina. J Mol Biol. 1969 Oct 28;45(2):397–406. [PubMed]
  • Hollyfield JG, Rayborn ME. Endocytosis in the inner segment of rod photoreceptors: analysis of Xenopus laevis retinas using horseradish peroxidase. Exp Eye Res. 1987 Nov;45(5):703–719. [PubMed]
  • Hopkins CR, Gibson A, Shipman M, Miller K. Movement of internalized ligand-receptor complexes along a continuous endosomal reticulum. Nature. 1990 Jul 26;346(6282):335–339. [PubMed]
  • Köhler G, Milstein C. Derivation of specific antibody-producing tissue culture and tumor lines by cell fusion. Eur J Immunol. 1976 Jul;6(7):511–519. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Liang CJ, Yamashita K, Muellenberg CG, Shichi H, Kobata A. Structure of the carbohydrate moieties of bovine rhodopsin. J Biol Chem. 1979 Jul 25;254(14):6414–6418. [PubMed]
  • Lippincott-Schwartz J, Yuan LC, Bonifacino JS, Klausner RD. Rapid redistribution of Golgi proteins into the ER in cells treated with brefeldin A: evidence for membrane cycling from Golgi to ER. Cell. 1989 Mar 10;56(5):801–813. [PubMed]
  • Lippincott-Schwartz J, Donaldson JG, Schweizer A, Berger EG, Hauri HP, Yuan LC, Klausner RD. Microtubule-dependent retrograde transport of proteins into the ER in the presence of brefeldin A suggests an ER recycling pathway. Cell. 1990 Mar 9;60(5):821–836. [PubMed]
  • Lodish HF, Kong N, Hirani S, Rasmussen J. A vesicular intermediate in the transport of hepatoma secretory proteins from the rough endoplasmic reticulum to the Golgi complex. J Cell Biol. 1987 Feb;104(2):221–230. [PMC free article] [PubMed]
  • MacKenzie D, Arendt A, Hargrave P, McDowell JH, Molday RS. Localization of binding sites for carboxyl terminal specific anti-rhodopsin monoclonal antibodies using synthetic peptides. Biochemistry. 1984 Dec 18;23(26):6544–6549. [PubMed]
  • Mangini NJ, Pepperberg DR. Immunolocalization of 48K in rod photoreceptors. Light and ATP increase OS labeling. Invest Ophthalmol Vis Sci. 1988 Aug;29(8):1221–1234. [PubMed]
  • Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed]
  • Matter K, Brauchbar M, Bucher K, Hauri HP. Sorting of endogenous plasma membrane proteins occurs from two sites in cultured human intestinal epithelial cells (Caco-2). Cell. 1990 Feb 9;60(3):429–437. [PubMed]
  • Melançon P, Glick BS, Malhotra V, Weidman PJ, Serafini T, Gleason ML, Orci L, Rothman JE. Involvement of GTP-binding "G" proteins in transport through the Golgi stack. Cell. 1987 Dec 24;51(6):1053–1062. [PubMed]
  • Misumi Y, Misumi Y, Miki K, Takatsuki A, Tamura G, Ikehara Y. Novel blockade by brefeldin A of intracellular transport of secretory proteins in cultured rat hepatocytes. J Biol Chem. 1986 Aug 25;261(24):11398–11403. [PubMed]
  • Mueller SC, Hubbard AL. Receptor-mediated endocytosis of asialoglycoproteins by rat hepatocytes: receptor-positive and receptor-negative endosomes. J Cell Biol. 1986 Mar;102(3):932–942. [PMC free article] [PubMed]
  • Nathans J, Weitz CJ, Agarwal N, Nir I, Papermaster DS. Production of bovine rhodopsin by mammalian cell lines expressing cloned cDNA: spectrophotometry and subcellular localization. Vision Res. 1989;29(8):907–914. [PubMed]
  • Nir I, Papermaster DS. Differential distribution of opsin in the plasma membrane of frog photoreceptors: an immunocytochemical study. Invest Ophthalmol Vis Sci. 1983 Jul;24(7):868–878. [PubMed]
  • Oprian DD, Molday RS, Kaufman RJ, Khorana HG. Expression of a synthetic bovine rhodopsin gene in monkey kidney cells. Proc Natl Acad Sci U S A. 1987 Dec;84(24):8874–8878. [PubMed]
  • Orci L, Malhotra V, Amherdt M, Serafini T, Rothman JE. Dissection of a single round of vesicular transport: sequential intermediates for intercisternal movement in the Golgi stack. Cell. 1989 Feb 10;56(3):357–368. [PubMed]
  • Papermaster DS, Dreyer WJ. Rhodopsin content in the outer segment membranes of bovine and frog retinal rods. Biochemistry. 1974 May 21;13(11):2438–2444. [PubMed]
  • Papermaster DS, Converse CA, Siuss J. Membrane biosynthesis in the frog retina: opsin transport in the photoreceptor cell. Biochemistry. 1975 Apr 8;14(7):1343–1352. [PubMed]
  • Papermaster DS, Schneider BG, Besharse JC. Vesicular transport of newly synthesized opsin from the Golgi apparatus toward the rod outer segment. Ultrastructural immunocytochemical and autoradiographic evidence in Xenopus retinas. Invest Ophthalmol Vis Sci. 1985 Oct;26(10):1386–1404. [PubMed]
  • Papermaster DS, Schneider BG, DeFoe D, Besharse JC. Biosynthesis and vectorial transport of opsin on vesicles in retinal rod photoreceptors. J Histochem Cytochem. 1986 Jan;34(1):5–16. [PubMed]
  • Peters KR, Palade GE, Schneider BG, Papermaster DS. Fine structure of a periciliary ridge complex of frog retinal rod cells revealed by ultrahigh resolution scanning electron microscopy. J Cell Biol. 1983 Jan;96(1):265–276. [PMC free article] [PubMed]
  • Rodriguez-Boulan E, Nelson WJ. Morphogenesis of the polarized epithelial cell phenotype. Science. 1989 Aug 18;245(4919):718–725. [PubMed]
  • Salminen A, Novick PJ. A ras-like protein is required for a post-Golgi event in yeast secretion. Cell. 1987 May 22;49(4):527–538. [PubMed]
  • Schmied R, Holtzman E. Involvement of the Golgi apparatus in sorting of materials to opposite ends of frog rod retinal photoreceptors. J Neurobiol. 1989 Apr;20(3):115–138. [PubMed]
  • Schneider BG, Kraig E. Na+, K(+)-ATPase of the photoreceptor: selective expression of alpha 3 and beta 2 isoforms. Exp Eye Res. 1990 Nov;51(5):553–564. [PubMed]
  • Schneider BG, Shyjan AW, Levenson R. Co-localization and polarized distribution of Na,K-ATPase alpha 3 and beta 2 subunits in photoreceptor cells. J Histochem Cytochem. 1991 Apr;39(4):507–517. [PubMed]
  • Ulmer JB, Palade GE. Targeting and processing of glycophorins in murine erythroleukemia cells: use of brefeldin A as a perturbant of intracellular traffic. Proc Natl Acad Sci U S A. 1989 Sep;86(18):6992–6996. [PubMed]
  • Urbani L, Simoni RD. Cholesterol and vesicular stomatitis virus G protein take separate routes from the endoplasmic reticulum to the plasma membrane. J Biol Chem. 1990 Feb 5;265(4):1919–1923. [PubMed]
  • Valtorta F, Jahn R, Fesce R, Greengard P, Ceccarelli B. Synaptophysin (p38) at the frog neuromuscular junction: its incorporation into the axolemma and recycling after intense quantal secretion. J Cell Biol. 1988 Dec;107(6 Pt 2):2717–2727. [PMC free article] [PubMed]
  • Vaughan DK, Fisher SK, Bernstein SA, Hale IL, Linberg KA, Matsumoto B. Evidence that microtubules do not mediate opsin vesicle transport in photoreceptors. J Cell Biol. 1989 Dec;109(6 Pt 1):3053–3062. [PMC free article] [PubMed]
  • Walworth NC, Novick PJ. Purification and characterization of constitutive secretory vesicles from yeast. J Cell Biol. 1987 Jul;105(1):163–174. [PMC free article] [PubMed]
  • Walworth NC, Goud B, Kabcenell AK, Novick PJ. Mutational analysis of SEC4 suggests a cyclical mechanism for the regulation of vesicular traffic. EMBO J. 1989 Jun;8(6):1685–1693. [PubMed]
  • Wandinger-Ness A, Bennett MK, Antony C, Simons K. Distinct transport vesicles mediate the delivery of plasma membrane proteins to the apical and basolateral domains of MDCK cells. J Cell Biol. 1990 Sep;111(3):987–1000. [PMC free article] [PubMed]
  • Young RW. Visual cells and the concept of renewal. Invest Ophthalmol Vis Sci. 1976 Sep;15(9):700–725. [PubMed]
  • Young RW, Droz B. The renewal of protein in retinal rods and cones. J Cell Biol. 1968 Oct;39(1):169–184. [PMC free article] [PubMed]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press