title: Postsynaptic response kinetics are controlled by a glutamate transporter at cone photoreceptors.
abstract: We evaluated the role of the sodium/glutamate transporter at the synaptic terminals of cone photoreceptors in controlling postsynaptic response kinetics. The strategy was to measure the changes in horizontal cell response rate induced by blocking transporter uptake in cones with dihydrokainate (DHK). DHK was chosen as the uptake blocker because, as we show through autoradiographic uptake measurements, DHK specifically blocked uptake in cones without affecting uptake in Mueller cells. Horizontal cells depolarized from about -70 to -20 mV as the exogenous glutamate concentration was increased from approximately 1 to 40 microM, so horizontal cells can serve as "glutamate electrodes" during the light response. DHK slowed the rate of hyperpolarization of the horizontal cells in a dose-dependent way, but didn't affect the kinetics of the cone responses. At 300 microM DHK, the rate of the horizontal cell hyperpolarization was slowed to only 17 +/- 8.5% (mean +/- SD) of control. Translating this to changes in glutamate concentration using the slice dose response curve as calibration in Fig. 2, DHK reduced the rate of removal of glutamate from approximately 0.12 to 0.031 microM/s. The voltage dependence of uptake rate in the transporter alone was capable of modulating glutamate concentration: we blocked vesicular released glutamate with bathed 20 mM Mg2+ and then added 30 microM glutamate to the bath to reestablish a physiological glutamate concentration level at the synapse and thereby depolarize the horizontal cells. Under these conditions, a light flash elicited a 17-mV hyperpolarization in the horizontal cells. When we substituted kainate, which is not transported, for glutamate, horizontal cells were depolarized but light did not elicit any response, indicating that the transporter alone was responsible for the removal of glutamate under these conditions. This suggests that the transporter was both voltage dependent and robust enough to modulate glutamate concentration. The transporter must be at least as effective as diffusion in removing glutamate from the synapse because there is only a very small light response once the transporter is blocked. The transporter, via its voltage dependence on cone membrane potential, appears to contribute significantly to the control of postsynaptic response kinetics.
authors.affiliation: Department of Molecular and Cell Biology, Division of Neurobiology, University of California at Berkeley, Berkeley, California 94720, USA.
authors.forename: L
authors.initials: L
authors.lastname: Gaal
authors.forename: B
authors.initials: B
authors.lastname: Roska
authors.forename: S A
authors.initials: SA
authors.lastname: Picaud
authors.forename: S M
authors.initials: SM
authors.lastname: Wu
authors.forename: R
authors.initials: R
authors.lastname: Marc
authors.forename: F S
authors.initials: FS
authors.lastname: Werblin
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000557:Ambystoma; D027322:Amino Acid Transport System X-AG; D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism; D001692:Biological Transport; D002118:Calcium / Q000494:pharmacology; D003624:Darkness; D018698:Glutamic Acid / Q000378:metabolism / Q000494:pharmacology; D007608:Kainic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D007700:Kinetics; D008027:Light; D008274:Magnesium / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects / Q000528:radiation effects; D018408:Patch-Clamp Techniques; D012160:Retina / Q000166:cytology / Q000502:physiology*; D017949:Retinal Cone Photoreceptor Cells / Q000166:cytology / Q000502:physiology*; D013569:Synapses / Q000187:drug effects / Q000502:physiology*; D009435:Synaptic Transmission / Q000187:drug effects / Q000502:physiology*
chemical_list: D018528:ATP-Binding Cassette Transporters; D027322:Amino Acid Transport System X-AG; D001224:Aspartic Acid; D018698:Glutamic Acid; C020889:dihydrokainic acid; D008274:Magnesium; D007608:Kainic Acid; D002118:Calcium