Skip to main content
The Journal of Physiology logoLink to The Journal of Physiology
. 1973 Dec;235(2):437–446. doi: 10.1113/jphysiol.1973.sp010395

Uptake of [3H]glycine and [14C]glutamate by cultures of chick spinal cord

Y D Cho, R O Martin, G Tunnicliff
PMCID: PMC1350754  PMID: 4797124

Abstract

1. Spinal cord explants from chick embryos, grown in culture for up to 16 days, rapidly accumulated [3H]glycine and [14C]glutamate when incubated at 25° C in a medium containing either 2 × 10-10 M glycine or 4·8 × 10-8 M glutamate.

2. After 90 min incubation, a tissue/medium ratio of 60:1 and 20:1 was attained for [14C]glutamate and [3H]glycine respectively.

3. The uptake systems, in addition to requiring sodium ions in the medium, were temperature sensitive, showed saturation kinetics, and were inhibited by ouabain.

4. For the glutamate and glycine accumulation the Km value was 4·3 × 10-5 and 4·1 × 10-5 M respectively, indicating that a high affinity uptake process is involved.

5. The rate of accumulation of both glutamate and glycine increased in cultures between the ages of 3 and 10 days thus matching their morphological development.

6. In light of previous evidence, the demonstration of an active transport mechanism for both glutamate and glycine in spinal-cord-cultures that also shows a relationship with morphological maturity, suggests that these two amino acids may play a major role in spinal cord function.

Full text

PDF
437

Selected References

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

  1. Balcar V. J., Johnston G. A. High affinity uptake of transmitters: studies on the uptake of L-aspartate, GABA, L-glutamate and glycine in cat spinal cord. J Neurochem. 1973 Feb;20(2):529–539. doi: 10.1111/j.1471-4159.1973.tb12152.x. [DOI] [PubMed] [Google Scholar]
  2. Bunge M. B., Bunge R. P., Peterson E. R. The onset of synapse formation in spinal cord cultures as studied by electron microscopy. Brain Res. 1967 Dec;6(4):728–749. doi: 10.1016/0006-8993(67)90129-1. [DOI] [PubMed] [Google Scholar]
  3. Crain S. M. Development of "organotypic" bioelectric activities in central nervous tissues during maturation in culture. Int Rev Neurobiol. 1966;9:1–43. doi: 10.1016/s0074-7742(08)60135-x. [DOI] [PubMed] [Google Scholar]
  4. Duggan A. W., Johnston G. A. Glutamate and related amino acids in cat spinal roots, dorsal root ganglia and peripheral nerves. J Neurochem. 1970 Aug;17(8):1205–1208. doi: 10.1111/j.1471-4159.1970.tb03369.x. [DOI] [PubMed] [Google Scholar]
  5. Graham L. T., Jr, Shank R. P., Werman R., Aprison M. H. Distribution of some synaptic transmitter suspects in cat spinal cord: glutamic acid, aspartic acid, gamma-aminobutyric acid, glycine and glutamine. J Neurochem. 1967 Apr;14(4):465–472. doi: 10.1111/j.1471-4159.1967.tb09545.x. [DOI] [PubMed] [Google Scholar]
  6. HILD W., TASAKI I. Morphological and physiological properties of neurons and glial cells in tissue culture. J Neurophysiol. 1962 Mar;25:277–304. doi: 10.1152/jn.1962.25.2.277. [DOI] [PubMed] [Google Scholar]
  7. Hosli E., Ljungdahl A., Hokfelt T., Hosli L. Spinal cord tissue cultures--a model for autoradiographic studies on uptake of putative neurotransmitters such as glycine and GABA. Experientia. 1972 Nov 15;28(11):1342–1344. doi: 10.1007/BF01965334. [DOI] [PubMed] [Google Scholar]
  8. Kim S. U., O T. H., Johnson D. D. Developmental changes of acetylcholinesterase and pseudocholinesterase in organotypic cultures of spinal cord. Exp Neurol. 1972 May;35(2):274–281. doi: 10.1016/0014-4886(72)90153-7. [DOI] [PubMed] [Google Scholar]
  9. Kim S. U. Observations on cerebellar granule cells in tissue culture. A silver and electron microscopic study. Z Zellforsch Mikrosk Anat. 1970;107(4):454–465. doi: 10.1007/BF00335434. [DOI] [PubMed] [Google Scholar]
  10. Levi G. Transport systems for GABA and for other amino acids in incubated chick brain tissue during development. Arch Biochem Biophys. 1972 Jul;151(1):8–21. doi: 10.1016/0003-9861(72)90466-3. [DOI] [PubMed] [Google Scholar]
  11. Logan W. J., Snyder S. H. Unique high affinity uptake systems for glycine, glutamic and aspartic acids in central nervous tissue of the rat. Nature. 1971 Dec 3;234(5327):297–299. doi: 10.1038/234297b0. [DOI] [PubMed] [Google Scholar]
  12. Neal M. J., Pickles H. G. Uptake of 14C glycine by spinal cord. Nature. 1969 May 17;222(5194):679–680. doi: 10.1038/222679a0. [DOI] [PubMed] [Google Scholar]
  13. Neal M. J. The uptake of [14C]glycine by slices of mammalian spinal cord. J Physiol. 1971 May;215(1):103–117. doi: 10.1113/jphysiol.1971.sp009460. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Seeds N. W. Biochemical differentiation in reaggregating brain cell culture. Proc Natl Acad Sci U S A. 1971 Aug;68(8):1858–1861. doi: 10.1073/pnas.68.8.1858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Tunnicliff G., Cho Y. D., Blackwell N., Martin R. O., Wood J. D. The uptake of gamma-aminobutyrate by organotypic cultures of chick spinal cord. Biochem J. 1973 May;134(1):27–32. doi: 10.1042/bj1340027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Tunnicliff G., Kim S. U. Synaptogenesis and the development of neurotransmitter enzymes in organotypic cultures of chick spinal cord. Brain Res. 1973 Jan 30;49(2):410–416. doi: 10.1016/0006-8993(73)90432-0. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Physiology are provided here courtesy of The Physiological Society

RESOURCES