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Volume 16, Number 16,
Issue of August 15, 1996
pp. 5060-5072
Copyright ©1996 Society for Neuroscience
Optical Recordings of the Effects of Cholinergic Ligands on
Neurons in the Ganglion Cell Layer of Mammalian Retina
Received April 10, 1996; revised May 22, 1996; accepted May 24, 1996.
William H. Baldridge
Vision, Touch, and Hearing Research Centre, Department of
Physiology and Pharmacology, The University of Queensland, Brisbane,
Queensland 4072, Australia
Cholinergic regulation of the activity of rabbit retinal ganglion
cells and displaced amacrine cells was investigated using optical
recording of changes in intracellular free calcium
([Ca2+]i). Labeling of
neurons in the mature retina was achieved by injecting calcium green-1
dextran (CaGD) into the isolated retina. Nicotine increased ganglion
cell [Ca2+]i, affecting every
loaded cell in some preparations; the pharmacology of nicotine was
consistent with an action at neuronal nicotinic receptors, and
specifically it was  -(neuronal-)bungarotoxin-sensitive but
 -bungarotoxin-insensitive. Muscarine also raised
[Ca2+]i, but it was less
potent than nicotine, affecting only a subpopulation of ganglion cells,
with an M1-like muscarinic receptor pharmacology. Neither the nicotine-
nor muscarine-induced increases of ganglion cell
[Ca2+]i were blocked by
the glutamate receptor antagonists 6,7-dinitroquinoxaline-2,3-dione and
aminophosphonopentanoic acid. Therefore, the effects of cholinergic
agonists on ganglion cell
[Ca2+]i were not
attributable to an indirect effect mediated by glutamatergic bipolar
cells. The effects of nicotine and muscarine were abolished in
calcium-free solution, indicating that the responses depend on calcium
influx.
Displaced (Cb) cholinergic amacrine cells were also loaded
with CaGD and were identified by selective labeling with the nuclear
dye 4 ,6-diamidino-2-phenyl-indole. Cb amacrine cells did
not respond to either nicotine or muscarine, but responded vigorously
to the glutamate receptor agonist kainic acid. There is anatomical
evidence indicating that cholinergic amacrine cells make synaptic
contact with each other, but the present results do not support the
hypothesis that communication between these cells is cholinergic.
Key words:
retinal ganglion cell;
amacrine cell;
intracellular
calcium;
acetylcholine;
calcium imaging;
receptors
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