Cell and Tissue Research

, Volume 269, Issue 1, pp 39–48 | Cite as

Morphogenesis of the photoreceptor outer segment during postnatal development in the mouse (BALB/c) retina

  • Shuichi Obata
  • Jiro Usukura


Disc formation of rod photoreceptor cells in developing BALB/c mice retinas was studied by rapid freeze, freeze-substitution, freeze-etching, immunocytochemistry, and myosin S-1 decoration methods. Freeze-substituted photoreceptor cells contained variously shaped vesicles in the apical swelling of the connecting cilium or the base of the outer segment during postnatal development. Rapid freezing successfully arrested pinocytosis; the fusion of small vesicles to give large ones, and the compression of certain vesicles (0.3–0.6 μm) appears to lead gradually to the formation of the so-called discs. We therefore propose that membranous discs are formed by the fusion of small pinocytotic vesicles and their subsequent compression. Discs formed in this way were partially stacked, but were ordered at random during the early developmental stages. During development, a partial stack of discs was progressively rearranged to a regular form as seen in mature outer segments. Cytoskeletal actin was expected to be involved in the disc formation; it was demonstrated in the distal axoneme of the connecting cilium during development and showed no change in its distribution. However, the polarity of the actin filaments, as revealed by myosin S-1 decoration in early developmental stages, was much more variable than in the adult. Barbed ends of actin filaments were associated with the plasma membrane or the membrane of vesicles. We also found actin filaments coiled up helically on ciliary microtubules.

Key words

Retina Photoreceptor cells Outer segment Disc membrane Morphogenesis Rapid freezing Actin immunocytochemistry Mouse (BALB/c) 


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  1. Albert B, Bray D, Lewis J, Raff M, Roberts K, Watson JD (1989) Molecular biology of the cell, 2nd edn. Garland, New York, pp 613–680Google Scholar
  2. Anderson DH, Fisher SK (1976) The photoreceptors of diurnal squirrels: outer segment structure, disc shedding, and protein renewal. J Ultrastruct Res 55:119–141PubMedGoogle Scholar
  3. Arikawa K, Williams DS (1989) Organization of actin filaments and immunocolocalization of alpha-actinin in the connecting cilium of rat photoreceptors. J Comp Neurol 288:640–646PubMedGoogle Scholar
  4. Chaitin MH (1991) Actin filaments in the photoreceptor cilium of the rds mutant mouse. Exp Eye Res 53:107–113CrossRefPubMedGoogle Scholar
  5. Chaitin MH, Bok D (1984) Immunoferritin localization of actin, myosin, and calmodulin in photoreceptor outer segments. J Cell Biol 99:114aGoogle Scholar
  6. Chaitin MH, Bok D (1986) Immunoferritin localization of actin in retinal photoreceptors. Invest Ophthalmol Vis Sci 27:1764–1767PubMedGoogle Scholar
  7. Chaitin MH, Burnside B (1989) Actin filament polarity at the site of rod outer segment disk morphogenesis. Invest Ophthalmol Vis Sci 30:2461–2469PubMedGoogle Scholar
  8. Chaitin MH, Schneider BG, Hall MO, Papermaster DS (1984) Actin in the photoreceptor connecting cilium: immunocytochemical localization to the site of outer segment disk formation. J Cell Biol 99:239–247CrossRefPubMedGoogle Scholar
  9. Chaitin MH, Carlsen RB, Samara GJ (1988) Immunogold localization of actin in developing photoreceptor cilia of normal and rds mutant mice. Exp Eye Res 47:437–446CrossRefPubMedGoogle Scholar
  10. Cohen Al (1961) The fine structure of the extrafoveal receptors of the rhesus monkey. Exp Eye Res 1:128–136PubMedGoogle Scholar
  11. Connell G, Bascom R, Molday L, Reid D, McInnes RR, Molday RS (1991) Photoreceptor peripherin is the normal product of the gene responsible for retinal degeneration in the rds mouse. Proc Natl Acad Sci USA 88:723–726PubMedGoogle Scholar
  12. De Robertis E (1956) Morphogenesis of the retinal rods; an electron microscope study. J Biophys Biochem Cytol 2:209–225PubMedGoogle Scholar
  13. De Robertis E (1960) Some observations on the ultrastructure and morphogenesis of photoreceptors. J Gen Physiol 43:1–13CrossRefGoogle Scholar
  14. Del Priore LV, Lewis A, Tan S, Carley WW, Webb WW (1987) Fluorescence light microscopy of F-actin in retinal rods and glial cells. Invest Ophthalmol Vis Sci 28:633–639PubMedGoogle Scholar
  15. Fliesler SJ, Rayborn ME, Hollyfield JG (1985) Membrane morphogenesis in retinal rod outer segments: inhibition by tunicamycin. J Cell Biol 100:574–587CrossRefPubMedGoogle Scholar
  16. Ishikawa H, Usukura J, Yamada E (1982) Application of cryomicrotomy to the rapid-freeze, deep-etch replica method for unfixed tissues and cells. J Electron Microsc 31:198–201Google Scholar
  17. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedGoogle Scholar
  18. Moody MF, Robertson JD (1960) The fine structure of some retinal photoreceptors. J Biophys Biochem Cytol 7:87–97PubMedGoogle Scholar
  19. Nilsson SEG (1964) Receptor cell outer segment development and ultrastructure of the disk membranes in the retina of the tadpole (Rana pipiens). J Ultrastruct Res 11:581–620PubMedGoogle Scholar
  20. Olney JW (1968) An electron microscopic study of synapse formation, receptor outer segment development, and other aspects of developing mouse retina. Invest Ophthalmol 7:250–268PubMedGoogle Scholar
  21. Papermaster DS, Schneider BG, Zorn MA, Kraehenbuhl JP (1978) Immunocytochemical localization of a large intrinsic membrane protein to the incisures and margins of frog rod outer segment disks. J Cell Biol 78:415–425CrossRefPubMedGoogle Scholar
  22. Perry SV (1955) Myosin adenosine triphosphatase: ATP+H2O→ ADP+H3PO4. In: Colowick SP, Kaplan NO (eds) Methods in enzymology, vol 2. Academic Press, New York, pp 582–588Google Scholar
  23. Steinberg RH, Fisher SK, Anderson DH (1980) Disc morphogenesis in vertebrate photoreceptors. J Comp Neurol 190:501–518PubMedGoogle Scholar
  24. Tokuyasu K, Yamada E (1959) The fine structure of the retina studied with the electron microscope. IV. Morphogenesis of outer segments of retinal rods. J Biophys Biochem Cytol 6:225–237PubMedGoogle Scholar
  25. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354PubMedGoogle Scholar
  26. Travis GH, Sutcliffe JG, Bok D (1991) The retinal degeneration slow (rds) gene product is a photoreceptor disc membrane-associated glycoprotein. Neuron 6:61–70CrossRefPubMedGoogle Scholar
  27. Usukura J, Bok D (1987) Changes in the localization and content of opsin during retinal development in the rds mutant mouse: immunocytochemistry and immunoassay. Exp Eye Res 45:501–515PubMedGoogle Scholar
  28. Usukura J, Akahori H, Takahashi H, Yamada E (1983) An improved device for rapid freezing using liquid helium. J Electron Microsc 32:180–185Google Scholar
  29. Vaughan DK, Fisher SK (1987) The distribution of F-actin in cells isolated from vertebrate retinas. Exp Eye Res 44:393–406PubMedGoogle Scholar
  30. Vaughan DK, Fisher SK (1989) Cytochalasin D disrupts outer segment disc morphogenesis in situ in rabbit retina. Invest Ophthalmol Vis Sci 30:339–342PubMedGoogle Scholar
  31. Weeds AG, Pope B (1977) Studies on the chymotryptic digestion of myosin. Effects of divalent cations on proteolytic susceptibility. J Mol Biol 111:129–157PubMedGoogle Scholar
  32. Williams DS, Linberg KA, Vaughan DK, Fariss RN, Fisher SK (1988) Disruption of microfilament organization and deregulation of disk membrane morphogenesis by cytochalasin D in rod and cone photoreceptors. J Comp Neurol 272:161–176PubMedGoogle Scholar
  33. Young RW (1967) The renewal of photoreceptor cell outer segments. J Cell Biol 33:61–72CrossRefPubMedGoogle Scholar
  34. Young RW, Bok D (1969) Participation of the retinal pigment epithelium in the rod outer segment renewal process. J Cell Biol 42:392–403CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • Shuichi Obata
    • 1
  • Jiro Usukura
    • 1
  1. 1.Department of AnatomyNagoya University, School of MedicineNagoyaJapan

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