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 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.]	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.	[Add color bar]