Noise analysis of the glutamate-activated current in photoreceptors.
The glutamate-activated current in photoreceptors has been attributed both to a sodium/glutamate transporter and to a glutamate-activated chloride channel. We have further studied the glutamate-activated current in single, isolated photoreceptors from the tiger salamander using noise analysis on whole-cell patch-clamp recordings. In cones, the current is generated by chloride channels with a single-channel conductance of 0.7 pS and an open lifetime of 2.4 ms. The number of channels per cell is in the range of 10,000-20,000. Activation of the channels requires the presence of both glutamate and sodium. The single-channel conductance and the open lifetime of the channel are independent of the external concentration of glutamate and sodium. External glutamate and sodium affect only the opening rate of the channels. D,L-Threo-3-hydroxyaspartate (THA), a glutamate-transport blocker, is shown to be a partial agonist for the channel. The single-channel conductance is the same regardless of whether glutamate or THA is the ligand, but the open lifetime of the channel is only 0.8 ms with THA as ligand. The glutamate-activated current in rods has a similar single-channel conductance (0.74 pS) and open lifetime (3 ms). We propose a kinetic model, consistent with these results, to explain how a transporter can simultaneously act both as a sodium/glutamate-gated chloride channel and a glutamate/sodium cotransporter.
Larsson
H P
HP
Department of Molecular and Cell Biology, University of California, Berkeley 94720, USA.
Picaud
S A
SA
Werblin
F S
FS
Lecar
H
H
D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D000557:Ambystoma; D027322:Amino Acid Transport System X-AG; D000818:Animals; D001224:Aspartic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D001693:Biological Transport, Active; D055592:Biophysical Phenomena; D001703:Biophysics; D002352:Carrier Proteins / Q000378:metabolism; D018118:Chloride Channels / Q000187:drug effects / Q000378:metabolism; D004553:Electric Conductivity; D050491:Glutamate Plasma Membrane Transport Proteins; D018698:Glutamic Acid / Q000378:metabolism*; D066298:In Vitro Techniques; D015640:Ion Channel Gating; D007700:Kinetics; D008024:Ligands; D008433:Mathematics; D008954:Models, Biological; D010786:Photoreceptor Cells / Q000187:drug effects / Q000378:metabolism*; D017948:Retinal Rod Photoreceptor Cells / Q000378:metabolism; D012964:Sodium / Q000378:metabolism / Q000494:pharmacology; D027981:Symporters
D018528:ATP-Binding Cassette Transporters; D027322:Amino Acid Transport System X-AG; D002352:Carrier Proteins; D018118:Chloride Channels; D050491:Glutamate Plasma Membrane Transport Proteins; D008024:Ligands; D027981:Symporters; C011274:3-hydroxyaspartic acid; D001224:Aspartic Acid; D018698:Glutamic Acid; D012964:Sodium