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Journal of Neuroscience, Vol 11, 2199-2208, Copyright © 1991 by Society for Neuroscience
Release of endogenous excitatory amino acids from ON-type bipolar cells isolated from the goldfish retina
M Tachibana and T Okada
Department of Psychology, Faculty of Letters, University of Tokyo, Japan.
Excitatory amino acids are presumed to be the transmitter of retinal
bipolar cells. However, one of the essential criteria for the
identification of the transmitter, its release from the cells upon
depolarization, has not been demonstrated. This article examines the
release of endogenous excitatory amino acids from bipolar cells and
correlates this release with the influx of Ca2+. Bipolar cells with a
large, bulblike axon terminal (ON-type cells with mixed inputs from rods
and cones) were enzymatically isolated from the goldfish retina. Horizontal
cells dissociated from the catfish retina were used as a probe of
excitatory amino acids, because these cells can detect submicromolar
concentrations of L-glutamate with high selectivity. An isolated bipolar
cell was closely apposed to a dissociated horizontal cell, and each cell
was voltage clamped by a patch pipette in the whole- cell clamp
configuration. When the bipolar cell was depolarized from - 60 mV to a
potential more positive than -40 mV using a 500-msec voltage pulse, an
outward current (greater than 20 pA) was recorded from the apposed
horizontal cell, which was maintained at +40 mV. The reversal potential of
the current induced by the substance released from bipolar cells (Irs) was
close to 0 mV and was almost identical to the responses evoked with
ionophoretically applied L-glutamate. Both reversal potentials were shifted
to the same, more negative value when the external Na+ was replaced with
choline. Furthermore, the Irs was suppressed reversibly by the application
of kynurenic acid, a glutamate antagonist. When the Ca current (ICa) of the
bipolar cell was blocked by Cd2+, the Irs also disappeared. The peak
amplitude of Irs was closely related to that of the ICa. We conclude that
mixed rod/cone ON- type bipolar cells of the goldfish retina release an
endogenous excitatory amino acid or a closely related compound in a Ca(2+)-
dependent manner.
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