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. 1979 Apr 1;81(1):215–228. doi: 10.1083/jcb.81.1.215

Freeze-fracture evidence for the presence of cholesterol in particle- free patches of basal disks and the plasma membrane of retinal rod outer segments of mice and frogs

PMCID: PMC2111529  PMID: 314450

Abstract

The freeze-fracture technique was used to examine the membranes of the photoreceptors of mice and frogs. Particle-free patches were found in the plasma membrane and basal disk membranes of the outer segments of both mice and frogs housed at room temperature, but not in frogs kept in a cold room. These patches were shown not to be artifacts of cryoprotection or fixation, and they persisted when fresh isolated outer segments were frozen by an ultrarapid method. They were also found to persist in mouse rods when retinas were incubated and subsequently fixed at temperatures up to 80 degrees C. Cholesterol was implicated as a significant component of the patches by the observation that, in the outer segments, pits, induced by treatment with the sterol- specific polyene antibiotic filipin, were present in and confined to the particle-free patches. That these lesions are not inherently limited to particle-free membrane areas was evident in the apical plasma membrane of the photoreceptor inner segments, where particles and pits were intermixed. Treatment with saponin, a surface-active agent which specifically complexes cholesterol, resulted in the disappearance of the particle-free patches. Patches were found in basal disks of both mouse and frog rods but not in older disks nearer the pigment epithelium, which indicates that changes occur in the composition of disk membranes and/or in the molecular ordering of their protein and lipid components during the early phase of their transit from the base towards the apex of the outer segment.

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Selected References

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  1. Anderson R. E., Maude M. B. Lipids of ocular tissues. 8. The effects of essential fatty acid deficiency on the phospholipids of the photoreceptor membranes of rat retina. Arch Biochem Biophys. 1972 Jul;151(1):270–276. doi: 10.1016/0003-9861(72)90497-3. [DOI] [PubMed] [Google Scholar]
  2. Anderson R. E., Risk M. Lipids of ocular tissues. IX. The phospholipids of frog photoreceptor membranes. Vision Res. 1974 Jan;14(1):129–131. doi: 10.1016/0042-6989(74)90127-8. [DOI] [PubMed] [Google Scholar]
  3. BANGHAM A. D., HORNE R. W. NEGATIVE STAINING OF PHOSPHOLIPIDS AND THEIR STRUCTURAL MODIFICATION BY SURFACE-ACTIVE AGENTS AS OBSERVED IN THE ELECTRON MICROSCOPE. J Mol Biol. 1964 May;8:660–668. doi: 10.1016/s0022-2836(64)80115-7. [DOI] [PubMed] [Google Scholar]
  4. Basinger S., Bok D., Hall M. Rhodopsin in the rod outer segment plasma membrane. J Cell Biol. 1976 Apr;69(1):29–42. doi: 10.1083/jcb.69.1.29. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bibb C., Young R. W. Renewal of fatty acids in the membranes of visual cell outer segments. J Cell Biol. 1974 May;61(2):327–343. doi: 10.1083/jcb.61.2.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bibb C., Young R. W. Renewal of glycerol in the visual cells and pigment epithelium of the frog retina. J Cell Biol. 1974 Aug;62(2):378–389. doi: 10.1083/jcb.62.2.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bownds D., Gordon-Walker A., Gaide-Huguenin A. C., Robinson W. Characterization and analysis of frog photoreceptor membranes. J Gen Physiol. 1971 Sep;58(3):225–237. doi: 10.1085/jgp.58.3.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brown M. F., Miljanich G. P., Dratz E. A. Interpretation of 100- and 360-MHz proton magnetic resonance spectra of retinal rod outer segment disk membranes. Biochemistry. 1977 Jun 14;16(12):2640–2648. doi: 10.1021/bi00631a010. [DOI] [PubMed] [Google Scholar]
  9. Brown M. F., Miljanich G. P., Dratz E. A. Proton spin-lattice relaxation of retinal rod outer segment membranes and liposomes of extracted phospholipids. Proc Natl Acad Sci U S A. 1977 May;74(5):1978–1982. doi: 10.1073/pnas.74.5.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Bunt A. H. Fine structure and radioautography of rabbit photoreceptor cells. Invest Ophthalmol Vis Sci. 1978 Feb;17(2):90–104. [PubMed] [Google Scholar]
  11. Cone R. A. Rotational diffusion of rhodopsin in the visual receptor membrane. Nat New Biol. 1972 Mar 15;236(63):39–43. doi: 10.1038/newbio236039a0. [DOI] [PubMed] [Google Scholar]
  12. Daemen F. J., De Grip W. J., Jansen P. A. Biochemical aspects of the visual process. XX. The molecular weight of rhodopsin. Biochim Biophys Acta. 1972 Jul 21;271(2):419–428. doi: 10.1016/0005-2795(72)90217-6. [DOI] [PubMed] [Google Scholar]
  13. Demel R. A., Jansen J. W., van Dijck P. W., van Deenen L. L. The preferential interaction of cholesterol with different classes of phospholipids. Biochim Biophys Acta. 1977 Feb 14;465(1):1–10. doi: 10.1016/0005-2736(77)90350-9. [DOI] [PubMed] [Google Scholar]
  14. Dewey M. M., Davis P. K., Blasie J. K., Barr L. Localization of rhodopsin antibody in the retina of the frog. J Mol Biol. 1969 Feb 14;39(3):395–405. doi: 10.1016/0022-2836(69)90134-x. [DOI] [PubMed] [Google Scholar]
  15. Duppel W., Dahl G. Effect of phase transition on the distribution of membrane-associated particles in microsomes. Biochim Biophys Acta. 1976 Mar 19;426(3):408–417. doi: 10.1016/0005-2736(76)90386-2. [DOI] [PubMed] [Google Scholar]
  16. FINE B. S., ZIMMERMAN L. E. OBSERVATIONS ON THE ROD AND CONE LAYER OF THE HUMAN RETINA. A LIGHT AND ELECTRON MICROSCOPIC STUDY. Invest Ophthalmol. 1963 Oct;2:446–459. [PubMed] [Google Scholar]
  17. Grant C. W., McConnell H. M. Glycophorin in lipid bilayers. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4653–4657. doi: 10.1073/pnas.71.12.4653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Haest C. W., Verkleij A. J., De Gier J., Scheek R., Ververgaert P. H., Van Deenen L. L. The effect of lipid phase transitions on the architecture of bacterial membranes. Biochim Biophys Acta. 1974 Jul 12;356(1):17–26. doi: 10.1016/0005-2736(74)90290-9. [DOI] [PubMed] [Google Scholar]
  19. Hall M. O., Bok D., Bacharach A. D. 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. doi: 10.1016/0022-2836(69)90114-4. [DOI] [PubMed] [Google Scholar]
  20. Hollyfield J. G., Basinger S. F. Cyclic metabolism of photoreceptor cells. Invest Ophthalmol Vis Sci. 1978 Feb;17(2):87–89. [PubMed] [Google Scholar]
  21. Hong K., Hubbell W. L. Lipid requirements for Rhodopsin regenerability. Biochemistry. 1973 Oct 23;12(22):4517–4523. doi: 10.1021/bi00746a033. [DOI] [PubMed] [Google Scholar]
  22. Höchli M., Hackenbrock C. R. Fluidity in mitochondrial membranes: thermotropic lateral translational motion of intramembrane particles. Proc Natl Acad Sci U S A. 1976 May;73(5):1636–1640. doi: 10.1073/pnas.73.5.1636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. James R., Branton D. Lipid- and temperature-dependent structural changes in Acholeplasma laidlawii cell membranes. Biochim Biophys Acta. 1973 Oct 25;323(3):378–390. doi: 10.1016/0005-2736(73)90183-1. [DOI] [PubMed] [Google Scholar]
  24. Jan L. Y., Revel J. P. Ultrastructural localization of rhodopsin in the vertebrate retina. J Cell Biol. 1974 Aug;62(2):257–273. doi: 10.1083/jcb.62.2.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kaplan M. W., Deffebach M. E. Birefringence measurements of structural inhomogeneities in Rana pipiens rod outer segments. Biophys J. 1978 Jul;23(1):59–70. doi: 10.1016/S0006-3495(78)85432-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. LaVail M. M. Rod outer segment disc shedding in relation to cyclic lighting. Exp Eye Res. 1976 Aug;23(2):277–280. doi: 10.1016/0014-4835(76)90209-8. [DOI] [PubMed] [Google Scholar]
  27. LaVail M. M. Rod outer segment disk shedding in rat retina: relationship to cyclic lighting. Science. 1976 Dec 3;194(4269):1071–1074. doi: 10.1126/science.982063. [DOI] [PubMed] [Google Scholar]
  28. Leeson T. S. Freeze-etch studies of rabbit eye. II. Outer segments of retinal photoreceptors. J Anat. 1971 Jan;108(Pt 1):147–157. [PMC free article] [PubMed] [Google Scholar]
  29. Leeson T. S. Rat retinal rods: freeze-fracture replication of outer segments. Can J Ophthalmol. 1970 Jan;5(1):91–107. [PubMed] [Google Scholar]
  30. Letellier L., Moudden H., Shechter E. Lipid and protein segregation in Escherichia coli membrane: morphological and structural study of different cytoplasmic membrane fractions. Proc Natl Acad Sci U S A. 1977 Feb;74(2):452–456. doi: 10.1073/pnas.74.2.452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Olive J., Recouvreur M. Differentiation of retinal rod disc membranes in mice. Exp Eye Res. 1977 Jul;25(1):63–74. doi: 10.1016/0014-4835(77)90247-0. [DOI] [PubMed] [Google Scholar]
  32. Papermaster D. S., Dreyer W. J. Rhodopsin content in the outer segment membranes of bovine and frog retinal rods. Biochemistry. 1974 May 21;13(11):2438–2444. doi: 10.1021/bi00708a031. [DOI] [PubMed] [Google Scholar]
  33. Papermaster D. S., Schneider B. G., Zorn M. A., Kraehenbuhl J. P. Immunocytochemical localization of opsin in outer segments and Golgi zones of frog photoreceptor cells. An electron microscope analysis of cross-linked albumin-embedded retinas. J Cell Biol. 1978 Apr;77(1):196–210. doi: 10.1083/jcb.77.1.196. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Pontus M., Delmelle M. Fluid lipid fraction in rod outer segment membrane. Biochim Biophys Acta. 1975 Aug 20;401(2):221–230. doi: 10.1016/0005-2736(75)90306-5. [DOI] [PubMed] [Google Scholar]
  35. Poo M. M., Cone R. A. Lateral diffusion of phodopsin in Necturus rods. Exp Eye Res. 1973 Dec 24;17(6):503–510. doi: 10.1016/0014-4835(73)90079-1. [DOI] [PubMed] [Google Scholar]
  36. Poo M., Cone R. A. Lateral diffusion of rhodopsin in the photoreceptor membrane. Nature. 1974 Feb 15;247(5441):438–441. doi: 10.1038/247438a0. [DOI] [PubMed] [Google Scholar]
  37. Raubach R. A., Nemes P. P., Dratz E. A. Chemical labeling and freeze-fracture studies on the localization of rhodopsin in the rod outer segment disk membrane. Exp Eye Res. 1974 Jan;18(1):1–12. doi: 10.1016/0014-4835(74)90038-4. [DOI] [PubMed] [Google Scholar]
  38. Saibil H., Chabre M., Worcester D. Neutron diffraction studies of retinal rod outer segment membranes. Nature. 1976 Jul 22;262(5566):266–270. doi: 10.1038/262266a0. [DOI] [PubMed] [Google Scholar]
  39. Speth V., Wunderlich F. Membranes of Tetrahymena. II. Direct visualization of reversible transitions in biomembrane structure induced by temperature. Biochim Biophys Acta. 1973 Feb 16;291(3):621–628. doi: 10.1016/0005-2736(73)90467-7. [DOI] [PubMed] [Google Scholar]
  40. Tillack T. W., Kinsky S. C. A freeze-etch study of the effects of filipin on liposomes and human erythrocyte membranes. Biochim Biophys Acta. 1973 Sep 27;323(1):43–54. doi: 10.1016/0005-2736(73)90430-6. [DOI] [PubMed] [Google Scholar]
  41. Verkleij A. J., Nauta I. L., Werre J. M., Mandersloot J. G., Reinders B., Ververgaert P. H., de Gier J. The fusion of abnormal plasma lipoprotein (LP-X) and the erythrocyte membrane in patients with cholestasis studied by electronmicroscopy. Biochim Biophys Acta. 1976 Jun 17;436(2):366–376. doi: 10.1016/0005-2736(76)90200-5. [DOI] [PubMed] [Google Scholar]
  42. Verkleij A. J., Ververgaert P. H., van Deenen L. L., Elbers P. F. Phase transitions of phospholipid bilayers and membranes of Acholeplasma laidlawii B visualized by freeze fracturing electron microscopy. Biochim Biophys Acta. 1972 Nov 2;288(2):326–332. doi: 10.1016/0005-2736(72)90253-2. [DOI] [PubMed] [Google Scholar]
  43. Wallace B. A., Engelman D. M. The planar distributions of surface proteins and intramembrane particles in Acholeplasma laidlawii are differentially affected by the physical state of membrane lipids. Biochim Biophys Acta. 1978 Apr 20;508(3):431–449. doi: 10.1016/0005-2736(78)90090-1. [DOI] [PubMed] [Google Scholar]
  44. Wattiaux-De Coninck S., Dubois F., Wattiaux R. Lateral phase separations and structural integrity of the inner membrane of rat-liver mitochondria. Effect of compression. Implications in the centrifugation of these organelles. Biochim Biophys Acta. 1977 Dec 15;471(3):421–435. doi: 10.1016/0005-2736(77)90047-5. [DOI] [PubMed] [Google Scholar]
  45. Wunderlich F., Wallach D. F., Speth V., Fischer H. Differential effects of temperature on the nuclear and plasma membranes of lymphoid cells. A study by freeze-etch electron microscopy. Biochim Biophys Acta. 1974 Nov 27;373(1):34–43. doi: 10.1016/0005-2736(74)90102-3. [DOI] [PubMed] [Google Scholar]
  46. Young R. W., Droz B. The renewal of protein in retinal rods and cones. J Cell Biol. 1968 Oct;39(1):169–184. doi: 10.1083/jcb.39.1.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Young R. W. The renewal of photoreceptor cell outer segments. J Cell Biol. 1967 Apr;33(1):61–72. doi: 10.1083/jcb.33.1.61. [DOI] [PMC free article] [PubMed] [Google Scholar]

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