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
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?).
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[image]
[graphic of brain, hippocampus, cartoon of neural cell culture] |
[Stimulate cells]
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[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 Erase red line in graph Add color bar |
[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.]
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0.3 sec stimulation Leave red line intact |
[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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[Add color bar]
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