Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1986 Mar;83(6):1945–1949. doi: 10.1073/pnas.83.6.1945

Voltage-gated and synaptic currents in rat Purkinje cells in dissociated cell cultures.

T Hirano, H Ohmori
PMCID: PMC323201  PMID: 2419913

Abstract

The electrical properties of rat Purkinje cells and synapses from granule cells were studied in dissociated cell cultures. To identify the cells we used an immunohistochemical method and recorded voltage-gated and synaptic currents with the patch-clamp technique (the whole-cell mode). Cultured Purkinje cells generated action potentials similar to those recorded from in vitro slices or in vivo preparations. Early Na, late Ca inward current, and K outward currents were distinguished by ion substitution under voltage clamp. We also recorded spontaneous synaptic currents in Purkinje cells cultured with granule cells. These synaptic currents reversed direction at the membrane potential of 2.5 mV, which was similar to currents induced by L-glutamate. Therefore, these are most likely excitatory synaptic currents from granule cells. Since these properties of Purkinje cells examined here are similar to those in situ, the cells in dissociated cell cultures offer a great opportunity to study biophysical properties of identified neurons in the central nervous system.

Full text

PDF
1945

Images in this article

1946Image
on p.1946
1946Image
on p.1946
1946Image
on p.1946
1946Image
on p.1946
1946Image
on p.1946
1946Image
on p.1946
1946Image
on p.1946

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Fukuda J., Kameyama M., Yamaguchi K. Breakdown of cytoskeletal filaments selectively reduces Na and Ca spikes in cultured mammal neurones. Nature. 1981 Nov 5;294(5836):82–85. doi: 10.1038/294082a0. [DOI] [PubMed] [Google Scholar]
  2. Gallo V., Ciotti M. T., Coletti A., Aloisi F., Levi G. Selective release of glutamate from cerebellar granule cells differentiating in culture. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7919–7923. doi: 10.1073/pnas.79.24.7919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Garson J. A., Beverley P. C., Coakham H. B., Harper E. I. Monoclonal antibodies against human T lymphocytes label Purkinje neurones of many species. Nature. 1982 Jul 22;298(5872):375–377. doi: 10.1038/298375a0. [DOI] [PubMed] [Google Scholar]
  4. Giotta G. J., Heitzmann J., Cohn M. Immunological identification of cerebellar cell lines. Brain Res. 1982 Jun;256(2):209–221. doi: 10.1016/0165-3806(82)90043-8. [DOI] [PubMed] [Google Scholar]
  5. Hackett J. T., Hou S. M., Cochran S. L. Glutamate and synaptic depolarization of Purkinje cells evoked by parallel fibers and by climbing fibers. Brain Res. 1979 Jul 13;170(2):377–380. doi: 10.1016/0006-8993(79)90118-5. [DOI] [PubMed] [Google Scholar]
  6. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  7. Hausen P., Dreyer C. The use of polyacrylamide as an embedding medium for immunohistochemical studies of embryonic tissues. Stain Technol. 1981 Sep;56(5):287–293. doi: 10.3109/10520298109067329. [DOI] [PubMed] [Google Scholar]
  8. KARNOVSKY M. J., ROOTS L. A "DIRECT-COLORING" THIOCHOLINE METHOD FOR CHOLINESTERASES. J Histochem Cytochem. 1964 Mar;12:219–221. doi: 10.1177/12.3.219. [DOI] [PubMed] [Google Scholar]
  9. Kornhuber J., Fischer E. G. Glutamic acid diethyl ester induces catalepsy in rats. A new model for schizophrenia? Neurosci Lett. 1982 Dec 31;34(3):325–329. doi: 10.1016/0304-3940(82)90196-3. [DOI] [PubMed] [Google Scholar]
  10. Llinás R., Sugimori M. Electrophysiological properties of in vitro Purkinje cell somata in mammalian cerebellar slices. J Physiol. 1980 Aug;305:171–195. doi: 10.1113/jphysiol.1980.sp013357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Nilaver G., Defendini R., Zimmerman E. A., Beinfeld M. C., O'Donohue T. L. Motilin in the Purkinje cell of the cerebellum. Nature. 1982 Feb 18;295(5850):597–598. doi: 10.1038/295597a0. [DOI] [PubMed] [Google Scholar]
  12. Sandoval M. E., Cotman C. W. Evaluation of glutamate as a neurotransmitter of cerebellar parallel fibers. Neuroscience. 1978;3(2):199–206. doi: 10.1016/0306-4522(78)90101-x. [DOI] [PubMed] [Google Scholar]
  13. Sellinger O. Z., Legrand J., Clos J., Ohlsson W. G. Unequal patterns of development of succinate-dehydrogenase and acetylcholinesterase in Purkinje cell bodies and granule cells isolated in bulk from the cerebellar cortex of the immature rat. J Neurochem. 1974 Dec;23(6):1137–1144. doi: 10.1111/j.1471-4159.1974.tb12210.x. [DOI] [PubMed] [Google Scholar]
  14. Valcana T., Hudson D., Timiras P. S. Effects of X-irradiation on the content of amino acids in the developing rat cerebellum. J Neurochem. 1972 Sep;19(9):2229–2232. doi: 10.1111/j.1471-4159.1972.tb05133.x. [DOI] [PubMed] [Google Scholar]
  15. Weber A., Schachner M. Maintenance of immunocytologically identified Purkinje cells from mouse cerebellum in monolayer culture. Brain Res. 1984 Oct 8;311(1):119–130. doi: 10.1016/0006-8993(84)91404-5. [DOI] [PubMed] [Google Scholar]
  16. Young A. B., Oster-Granite M. L., Herndon R. M., Snyder S. H. Glutamic acid: selective depletion by viral induced granule cell loss in hamster cerebellum. Brain Res. 1974 Jun 14;73(1):1–13. doi: 10.1016/0006-8993(74)91002-6. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES