Advertisement

Neurochemical Research

, Volume 13, Issue 7, pp 625–631 | Cite as

Axoplasmic RNA species synthesized in the isolated squid giant axon

  • M. V. Rapallino
  • A. Cupello
  • A. Giuditta
Original Articles

Abstract

Isolated squid stellate nerves and giant fiber lobes were incubated for 8 hr in Millipore filtered sea water containing [3H]uridine. The electrophoretic patterns of radioactive RNA purified from the axoplasm of the giant axon and from the giant fiber lobe (cell bodies of the giant axon) demonstrated the presence of RNA species with mobilities corresponding to tRNA and rRNA. The presence of labeled rRNAs was confirmed by the behavior of the large rRNA component (31S) which, in the squid, readily dissociates into its two constituent moyeties (17S and 20S). Comparable results were obtained with the axonal sheath and the stellate nerve. In all the electrophoretic patterns, additional species of radioactive RNA migrated between the 4S and the 20S markers, i.e. with mobilities corresponding to presumptive mRNAs. Chromatographic analysis of the purified RNAs on oligo(dT)cellulose indicated the presence of labeled poly(A)+ RNA in all tissue samples. Radioactive poly(A)+ RNA represented approximately 1% of the total labeled RNA in the axoplasm, axonal sheath and stellate nerve, but more than 2% in the giant fiber lobe. The labeled poly(A)+ RNAs of the giant fibre lobe showed a prevalence of larger species in comparison to the axonal sheath and stellate nerve. In conclusion, the axoplasmic RNAs synthesized by the isolated squid giant axon appear to include all the major classes of axoplasmic RNAs, that is rRNA, tRNA and mRNA.

Key Words

Axon RNA RNA synthesis squid 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Koenig, E. 1984. Local synthesis of axonal protein. Pages 315–340,in Lajtha A. (ed.) Handbook of Neurochemistry, 2nd edition, Plenum Press, New York, vol. 7.Google Scholar
  2. 2.
    Ingoglia, N. A. 1979. 4S RNA is present in regenerating optic axons of goldfish. Science 206:73–75.Google Scholar
  3. 3.
    Koenig, E. 1979. Ribosomal RNA in Mauthner axon: implications for a protein synthesizing machinery in the myelinated axon. Brain Res. 174:95–107.Google Scholar
  4. 4.
    Lasek, R. J., Dabrowski, C., and Nordlander, R. 1973. Analysis of axoplasmic RNA from invertebrate giant axons. Nature [New Biol] 244:162–165.Google Scholar
  5. 5.
    Giuditta, A., Hunt, T., and Santella, L. 1986. messenger RNA in squid axoplasm. Neurochem. Intern. 8:435–442.Google Scholar
  6. 6.
    Giuditta, A., Hunt, T., Perrone Capano, C., Santella, L., and Kaplan, B. B. 1986. Messenger RNA is present in the axoplasm of squid giant axons. Pages 42–56, in Giuditta, A., Kaplan B. B. and Zomzely Neurath C. (eds.) Role of RNA and DNA in Brain Function. A Molecular Biological Approach. Martinus Nijhoff Publ., Boston.Google Scholar
  7. 7.
    Giuditta, A., Cupello, A., and Lazzarini, G. 1980. Ribosomal RNA in the axoplasm of the squid giant axon. J. Neurochem. 34:1757–1760.Google Scholar
  8. 8.
    Giuditta, A. 1980. Origin of axoplasmic protein in the squid giant axon. Riv. Biol. 73:35–49.Google Scholar
  9. 9.
    Koenig, E. 1967. Synthetic mechanisms in the axon—IV. In vitro incorporation of [3H]precursors into axonal protein and RNA. J. Neurochem. 14:437–446.Google Scholar
  10. 10.
    Edström, A., Edström, J.-E., and Hökfelt, T. 1969. Sedimentation analysis of ribonucleic acid extracted from isolated Mauthner nerve fibre components. J. Neurochem. 16:53–66.Google Scholar
  11. 11.
    Cutillo, V., Montagnese, P., Gremo, F., Casola, L., and Giuditta, A. 1983. Origin of axoplasmic RNA in the squid giant fiber. Neurochem. Res. 8:1621–1634.Google Scholar
  12. 12.
    Giuditta, A., Dettbarn, W.-D., and Brzin, M. 1968. Protein synthesis in the isolated giant axon of the squid. Proc. Natl. Acad. Sci. USA 59:1284–1287.Google Scholar
  13. 13.
    Cupello, A., and Hydén, H. 1975 a. Fractionation of RNA from brain synaptosomes and cytoplasmic subcellular fractions. J. Neurochem. 25:399–406.Google Scholar
  14. 14.
    Cupello, A., and Hydén, H. 1975 b. Separation of RNA by microelectrophoresis in agarose-acrylamide gels. Neurobiol. 5:129–136.Google Scholar
  15. 15.
    Loenig, U. E. 1969. The determination of molecular weight of ribonucleic acid by polyacrylamide gel electrophoresis. Biochem. J. 113:131–138.Google Scholar
  16. 16.
    Cupello, A., Ferrillo, F., and Rosadini, G. 1979. Pattern of labelling of rabbit brain cortex poly(A)-associated RNA after a single electroconvulsive shock. Exp. Neurol. 63:451–457.Google Scholar
  17. 17.
    Cupello, A., and Hydén, H. 1976. Alterations of the pattern of hippocampal nerve cell RNA labelling during training in rats. Brain Res. 114:453–460.Google Scholar
  18. 18.
    Cammarano, P., Londei, P., and Mazzei, F. 1980. Physicochemical characterization of the ribosomal RNA species of the Mollusca. Biochem. J. 189:313–335.Google Scholar
  19. 19.
    Belmonte, F., Cupello, A., Lazzarini, G., and Giuditta, A. 1979. Electrophoretic characterization of ribosomal RNA in the squid. Comp. Biochem. Physiol. 63B:373–377.Google Scholar
  20. 20.
    Perrone Capano, C., Giuditta, A., Castigli, E., and Kaplan, B. B. 1987. Occurrence and sequence complexity of polyadenylated RNA in squid axoplasm. J. Neurochem. 49:698–704.Google Scholar
  21. 21.
    Alemà, S., and Giuditta, A. 1976. Site of biosynthesis of brain specific proteins in the giant fibre system of the squid. J. Neurochem. 26:995–999.Google Scholar
  22. 22.
    Ingoglia, N. A., Giuditta, A., Zanakis, M. F., Babigian, A., Tasaki, I., Chakraborty, G., and Sturman, J. 1983. Incorporation of3H-amonoacids into proteins in a partially purified fraction of axoplasm: evidence for transfer RNA mediated, post-translational protein modification in squid giant axon. J. Neurosci. 3:2463–2473.Google Scholar
  23. 23.
    Giuditta, A., Menichini, E., Castigli, E., Perrone Capano, C., Gioio, A., and Kaplan, B. B. 1986. Active polysomes are present in the axoplasm of the squid giant axon. Abstr. 2° Congr. Soc. Ital. Neurosc., 315.Google Scholar
  24. 24.
    Giuditta, A., Metafora, S., Felsani, A., and Del Rio, A. 1977. Factors for protein synthesis in the axoplasm of squid giant axons. J. Neurochem. 28:1393–1395.Google Scholar
  25. 24.
    Lasek, R. J., Gainer, H., and Barker, J. L. 1977. Cell to cell transfer of glial proteins to the squid giant axon. The glianeuron protein transfer hypothesis. J. Cell Biol. 74:501–523.Google Scholar
  26. 26.
    Hydén, H., and Pigon, A. 1960. A cytophysiological study of the functional relationship between oligodendroglial cells and nerve cells of Deiters' nucleus. J. Neurochem. 6:57–72.Google Scholar
  27. 27.
    Egyházy, E. and Hydén, H. 1961. Experimentally induced changes in the base composition of the ribonucleic acids of isolated nerve cells and their oligodendroglial cells. J. Biophys. Biochem. Cytol. 10:403–410.Google Scholar
  28. 28.
    Hydén, H., and Lange, P. W. 1966. A genetic stimulation with production of adenine-uracil rich RNA in neurons and glia in learning. The question of transfer of RNA from glia to neurons. Naturwiss. 53:64–70.Google Scholar
  29. 29.
    Kuffler, S. W., and Nicholls, J. G. 1966. The physiology of neuroglial cells. Ergebn. Physiol. 57:1–90.Google Scholar
  30. 30.
    Pevzner, L. Z. 1965. Topochemical aspects of nucleic acid and protein metabolism within the neuron-neuroglia unit of the superior cervical ganglion. J. Neurochem. 12:993–1002.Google Scholar

Copyright information

© Plenum Publishing Corporation 1988

Authors and Affiliations

  • M. V. Rapallino
    • 1
  • A. Cupello
    • 1
  • A. Giuditta
    • 2
  1. 1.C.N.R. Unit for Brain NeurophysiologyGenova
  2. 2.Department of General and Environmental PhysiologyUniversity of Naples and International Institute of Genetics and BiophysicsNaplesItaly

Personalised recommendations