Home Abstract Introduction
Construction of glutamate sensors Initial testing of sensors in vitro Optimization of sensor characteristics Expression of sensor in living cells Glutamate measurement in dendrites Glutamate measurement in response to frequency of action potentials
Glutamate function Sensor design
Protein purification through His tags Overexpression of protein using pRSETB Directing proteins to a membrane by signal peptides Transmembrane regions of proteins Expression of proteins on the surface of mammalian cells through pDISPLAY

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 response to frequency of action potentials

[Nerves may fire at different frequencies. How does that affect the concentration of spillover glutamate?

Three conditions:
- 1 action potential
- 10 action potentials spaced by 67 msec
- 10 action potentials spaced by 33 msec

Glutamate accumulates to higher level when time of reuptake is smaller.]


[Erase lines with open symbols. Label lines: black = 1 action potential; red = 10 action potentials, 1 per 67 msec; blue = 10 action potentials, 1 per 33 msec.]
[The idea that the difference in glutamate levels is due to a shorter period of reuptake can be tested by blocking reuptake with TBOA.

When reuptake blocked, magnitude of glutamate spike for 10 action potentials no longer depends on frequency]


[Erase lines with closed symbols. Label lines]