Journal of Comprehensible Results

Hires SA, Zhu Y, Tsien R (2008)
Optical measurement of synaptic glutamate spillover and reuptake
by linker optimized glutamate-sensitive fluorescent reporters
Proc Natl Acad Sci USA 105:4411-4416

(Translated by Jeff Elhai)

Experiment: Glutamate measurement in dendrites

[Transfect membrane-bound sensor gene into neuron cells from rat hippocampus (significance of hippocampus?).
[image]
[graphic of brain, hippocampus, cartoon of neural cell culture]
[Stimulate cells]
[image]
[graphic of external voltage set up]
[CFP/YFP fluorescence from Membrane-bound glutamate sensor in dendrites stimulated for 0.3 seconds (10 action potentials). Record before, during, after. Transient increase in CFP/YFP => transient increase in glutamate.

Quantitated and averaged over all dendrite surface.


0.3 sec stimulation

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[Concentration of neurotransmitter affected not only by release but also reuptake. Cocaine works largely by inhibiting the reuptake of the neurotransmitter dopamine.

[+/- inhibitor of glutamate reuptake: DL-threo-β-benzyloxyaspartate (TBOA)]

[image]
[cartoon of neurotransmitter reuptake, +/- drug that blocks reuptake]
[Same conditions as before, but with TBOA added to block glutamate reuptake. Glutamate spike persists.]

0.3 sec stimulation

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[Same as above, but for just one action potential (averaged over 30 separate repetitions). No TBOA. Show's mostly over after 0.1 seconds. Glutamate transient.

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