Skip to main content
Log in

Immunocytochemical localization of a rhodopsin-like protein in the lipochondria in photosensitive neurons of Aplysia californica

  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Summary

Polyclonal antibodies directed against squid opsin were used in immunocytochemical and immunoblot experiments to identify a rhodopsin-like protein in photosensitive neurons of Aplysia. Aldehyde-fixed abdominal and cerebral ganglia were embedded in paraffin for peroxidase anti-peroxidase analysis or used whole for immunofluorescence studies. Ganglia were embedded in Lowicryl K4M for electron-microscope immunocytochemistry. In both the cerebral and abdominal ganglia, light-microscope immunocytochemical results showed reaction product deposited around the neuronal cell periphery corresponding in position to the lipochondria. In the abdominal ganglion, the giant cell R2, located in the right rostral quarter, and neurons in the right caudal quarter were consistently labeled with anti-opsin. Electron-microscopic studies demonstrated ferritin-labeling of the lipochondria in R2 and other immunoreactive neurons. Immunoblot analysis of R2 and cerebral neuron extracts was used to identify two prominent immunoreactive protein bands at 85000 and 67500 molecular weight.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Andresen MC, Brown AM (1976) Voltage-clamp studies on the light response of identified interneuron L10. Biophys Soc Abstr

  • Arvanitaki A, Chalazonitis N (1961) Excitatory and inhibitory processes initiated by light and infra-red radiations in single identifiable nerve cells. In: Florey E (ed) Nervous inhibition. Pergamon, New York, pp 194–231

    Google Scholar 

  • Baur PS, Brown AM, Rogers TD, Brower ME (1977) Lipochondria and the light response of Aplysia giant neurons. J Neurobiol 8:19–42

    Google Scholar 

  • Benjamin PR, Walker TS (1972) Two pigments in the brain of a fresh-water pulmonate snail. Comp Biochem Physiol 418:813–821

    Google Scholar 

  • Bok D, Dockstader J, Horwitz J (1982) Immunocytochemical localization of the lens main intrinsic polypeptide (MIP) in communicating junctions. J Cell Biol 92:213–220

    Google Scholar 

  • Bok D, Ong DE, Chytil F (1984) Immunocytochemical localization of cellular retinol binding protein in the rat retina. Invest Ophthal Vis Sci 25:877–883

    Google Scholar 

  • Brown HM, Brown AM (1972) Ionic basis of the photoresponse of Aplvsia giant neuron: K+ permeability increase. Science 178:755–756

    Google Scholar 

  • Brown AM, Brown HM (1973) Light response of a giant Aplysia neurone. J Gen Physiol 62:239–254

    Google Scholar 

  • Brown AM, Baur PS, Tuley FH (1975) Phototransduction in Aplysia neurons: Calcium release from pigmented granules is essential. Science 188:157–160

    Google Scholar 

  • Brown AM, Brodwick MS, Eaton D (1977) Intracellular calcium and extra-retinal photoreception in Aplysia giant neurons. J Neurobiol 8:1–18

    Google Scholar 

  • Coggeshall RE (1967) A light and electron microscope study of the abdominal ganglion of Aplysia californica. J Neurophysiol 30:1263–1287

    Google Scholar 

  • Fitzgerald PG, Bok D, Horwitz J (1983) Immunocytochemical localization of the main intrinsic polypeptide (MIP) in ultrathin frozen sections of rat lens. J Cell Biol 97:1491–1499

    Google Scholar 

  • Foster KW, Sarnak J, Patel N, Zarilli G, Okabe M, Kline T, Nakanishi K (1984) A rhodopsin is the functional photoreceptor for phototaxis in the unicellular eucaryote Chlamydomonas. Nature 311:756–759

    Google Scholar 

  • Foster RG, Follett BK, Lythgoe JN (1985) Rhodopsin-like sensitivity of extra-retinal photoreceptors mediating the photoperiodic response in quail. Nature 313:50–52

    Google Scholar 

  • Hara T, Hara R (1972) Cephalopod retinochrome. In: Dartnall HJA (ed) Handbook of sensory physiology. Vol vii/l. Springer, New York, pp 720–746

    Google Scholar 

  • Hara T, Hara R (1980) Retinochrome and rhodopsin in the extraocular photoreceptor of the squid. J Gen Physiol 75:1–19

    Google Scholar 

  • Henkart M (1975) Light-induced changes in the structure of pigmented granules in Aplysia neurons. Science 188:155–157

    Google Scholar 

  • Herman E, Shannon LM (1984) Immunocytochemical evidence for the involvement of Golgi apparatus in the deposition of seed lectin of Bauhinia purpurea (Leguminosae). Protoplasma 121:163–170

    Google Scholar 

  • Kennedy DJ (1960) Neuronal photoreception in the lamelibranch mollusc. Gen Physiol 44:277–299

    Google Scholar 

  • Kito Y, Naito T, Nashima K (1982) Purification of squid and octopus rhodopsin. In: Packer L (ed) Methods in enzymology. Vol 81. Biomembranes; Part H: Visual pigments and purple membranes I. Academic Press, New York, pp 161–171

    Google Scholar 

  • Krauhs JM, Sordahl LA, Brown AM (1977) Isolation of pigmented granules involved in extra-retinal photoreception in Aplysia californica neurons. Biochem Biophys Acta 471:25–31

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • McAllister LB, Scheller RH, Kandel ER, Axel R (1983) In situ hybridization to study the origin and fate of identified neurons. Science 222:800–808

    Google Scholar 

  • McReynolds JS, Gorman ALF (1974) Ionic basis of hyperpolarizing receptor potential in scallop eye: Increase in permeability to potassium ions. Science 193:658–659

    Google Scholar 

  • Moore RC, Lalicker CG, Gisher AG (1952) Invertebrate fossils. McGraw-Hill Book Co, New York

    Google Scholar 

  • Morton JE (1960) Molluscs. Harper & Brothers, New York

    Google Scholar 

  • Nishioka RS, Yasumasu I, Bern HA (1966a) Photoreceptive features of vesicles associated with the nervous system of cephalopods. Nature 211:1181

    Google Scholar 

  • Nishioka RS, Yasumasu I, Packard A, Bern HA, Young JZ (1966b) Nature of vesicles associated with the nervous system of cephalopods. Z Zellforsch 75:301–316

    Google Scholar 

  • Rubles LJ, Cabebe CS, Aguilo JA, Anyakora PA, Bok D (1984) Autoradiographic and biochemical analysis of photoreceptor membrane renewal in Octopus retina. J Neurocytol 13:145–164

    Google Scholar 

  • Saibil HR, Michel-Villaz M (1984) Squid rhodopsin and GTP binding protein crossreact with vertebrate photoreceptor enzymes. Proc Natl Acad Sci USA 81:5111–5115

    Google Scholar 

  • Sternberger LA (1979) Immunocytochemistry. John Wiley & Sons, New York, pp 122–127

    Google Scholar 

  • Towbin H, Staehlin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Sci USA 76:4350–4354

    Google Scholar 

  • Vandenberg CA, Montai M (1984) Light-regulated biochemical events in invertebrate photoreceptors. I. Light-activated guanosinetriphosphatase, guanine nucleotide binding, and cholera toxin catalyzed labeling of squid photoreceptor membranes. Biochemistry 23:2339–2347

    Google Scholar 

  • Zolotov VV (1982) Spatial organization of the receptive fields in light sensitive neurons of the abdominal nerve cord in insects. Zh Evol Biokhim Fiziol 18:366–372

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Robles, L.J., Breneman, J.W., Anderson, E.O. et al. Immunocytochemical localization of a rhodopsin-like protein in the lipochondria in photosensitive neurons of Aplysia californica . Cell Tissue Res. 244, 115–120 (1986). https://doi.org/10.1007/BF00218388

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00218388

Key words

Navigation