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Journal of Neuroscience, Vol 15, 3571-3582, Copyright © 1995 by Society for Neuroscience
L-glutamate-induced responses and cGMP-activated channels in three subtypes of retinal bipolar cells dissociated from the cat
P de la Villa, T Kurahashi and A Kaneko
Department of Information Physiology, National Institute for Physiological Sciences, Okazaki, Japan.
Effects of L-glutamate (Glu), the neurotransmitter released by
photoreceptors, on isolated cat bipolar cells were examined. Membrane
currents of bipolar cells were recorded by the patch-clamp technique in a
conventional whole-cell recording configuration using pipettes containing 1
mM cGMP, which has been known to activate a cationic current sensitive to
Glu in ON-type bipolar cells. ON-type bipolar cells (depolarized by light
in in situ) and OFF-type bipolar cells (hyperpolarized by light) were
identified by their response polarity to Glu. When the whole-cell
configuration was established, ON-type bipolar cells showed a steady inward
current which was suppressed by Glu, consistent with the response polarity
observed in in situ recordings. In contrast, OFF-type cells did not show a
steady current during the recordings. However, they responded to Glu with
an increase in cationic conductance. Among recorded cells, rod-driven
bipolar cells were identified by their immunoreactivity to anti-protein
kinase C (PKC-IR) antibody. Examination of PKC-IR revealed that ON-type
bipolar cells included both rod- and cone-driven bipolar cells, while
OFF-type cells were all cone-driven bipolar cells. The cGMP-activated
current observed in ON-type cells was accompanied by a change in the
current fluctuation due to the opening and closing of underlying channels.
Fluctuation analysis gave a unitary conductance value of 13 pS. In half of
the cells examined, maximum open probability reached almost 100%. The cGMP-
activated channel in bipolar cells seems novel, fundamentally different
from those found in photoreceptor cells or olfactory receptor cells.
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