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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
Author(s) -
Fosse Viggo M.,
Kolstad Jens,
Fonnum Frode
Publication year - 1986
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1986.tb04507.x
Subject(s) - glutamate receptor , superior colliculus , chemistry , bioluminescence , cortex (anatomy) , visual cortex , biochemistry , brain cortex , glutamic acid , biophysics , inferior colliculus , nucleus , neuroscience , biology , endocrinology , receptor , amino acid
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.