A Bioluminescence Method for the Measurement of l‐Glutamate: Applications to the Study of Changes in the Release of l‐Glutamate from Lateral Geniculate Nucleus and Superior Colliculus After Visual Cortex Ablation in Rats

A Bioluminescence Method for the Measurement of l‐Glutamate: Applications to the Study of... Abstract: We have developed a rapid, simple, specific, and very sensitive bioluminescence method for the measurement of l‐glutamate (l‐Glu). Oxidation of l‐Glu by glutamate dehydrogenase has been coupled with bacterial FMN reductase and luciferase. Light production (i.e., peak height or integral) was linear from < 0.5 to 500 pmol of l‐Glu. Potential interfering substances that may be encountered in brain tissue have been identified. The most potent inhibitors were ascorbate and the biogenic amines. Procedures that conferred long‐term stability of the reagent mixture (> 8 h) were established. Bioluminescence analysis of L‐Glu content in brain tissue extracts, fractions from release experiments, and human CSF corroborated respective results obtained by HPLC analysis. In this study, we have applied the method to monitor changes in the KCl‐evoked release of endogenous L‐Glu from milligram amounts of brain tissue, i.e., from lateral geniculate nucleus and superior colliculus after visual cortex ablation. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Neurochemistry Wiley

A Bioluminescence Method for the Measurement of l‐Glutamate: Applications to the Study of Changes in the Release of l‐Glutamate from Lateral Geniculate Nucleus and Superior Colliculus After Visual Cortex Ablation in Rats

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Publisher
Wiley
Copyright
Copyright © 1986 Wiley Subscription Services, Inc., A Wiley Company
ISSN
0022-3042
eISSN
1471-4159
D.O.I.
10.1111/j.1471-4159.1986.tb04507.x
Publisher site
See Article on Publisher Site

Abstract

Abstract: We have developed a rapid, simple, specific, and very sensitive bioluminescence method for the measurement of l‐glutamate (l‐Glu). Oxidation of l‐Glu by glutamate dehydrogenase has been coupled with bacterial FMN reductase and luciferase. Light production (i.e., peak height or integral) was linear from < 0.5 to 500 pmol of l‐Glu. Potential interfering substances that may be encountered in brain tissue have been identified. The most potent inhibitors were ascorbate and the biogenic amines. Procedures that conferred long‐term stability of the reagent mixture (> 8 h) were established. Bioluminescence analysis of L‐Glu content in brain tissue extracts, fractions from release experiments, and human CSF corroborated respective results obtained by HPLC analysis. In this study, we have applied the method to monitor changes in the KCl‐evoked release of endogenous L‐Glu from milligram amounts of brain tissue, i.e., from lateral geniculate nucleus and superior colliculus after visual cortex ablation.

Journal

Journal of NeurochemistryWiley

Published: Aug 1, 1986

References

  • Glutamate: a neurotransmitter in mammalian brain
    Fonnum, Fonnum
  • Application to mammalian tissues of the chemiluminescent method for detecting acetylcholine
    Israel, Israel; Lesbats, Lesbats
  • Release of endogenous amino acids from superior colliculus of the rabbit: in vitro studies after retinal ablation
    Sandberg, Sandberg; Corrazzi, Corrazzi
  • Release of endogenous and accumulated exogenous amino acids from slices of normal and climbing fibre‐deprived rat cerebellar slices
    Toggenburger, Toggenburger; Wiklund, Wiklund; Henke, Henke; Cuenod, Cuenod
  • Quantitative methods for measuring the histochemical distribution of alanine, glutamate and glutamine in brain
    Young, Young; Lowry, Lowry

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