In the isolated, perfused retina of the carp, properties of the feedback pathway from horizontal cells to cones were investigated by means of electrophysiological and neuropharmacological methods. When horizontal cells were hyperpolarized by illumination with an annulus, a depolarizing synaptic potential was produced in cones at the center of the annulus, suggesting that horizontal cells receive inputs from cones and exert a negative feedback to cones. On the other hand, a hyperpolarizing response (IPSP) was detected in cones, when a transient depolarization (EPSP) was induced in horizontal cells by application of a transretinal current pulse, which activated the release of transmitter from the photoreceptor terminals. The IPSP was abolished when the retina was perfused with a GABA-containing Ringer solution, because of desensitization of the feedback synapse. GABA also hyperpolarized the cone membrane, indicating the presence of a GABA-sensitive site in the cone. These results suggest that the GABA-mediated negative feedback operates from horizontal cells to cones in the dark, and ceases its function in the light.

SUMMARY1. Amacrine and ganglion cells in the carp retina were identified from such criteria as photoresponses, intracellular dye staining, responses to optic nerve stimulation and behaviour to a synapse blocking agent.2. Responses of ganglion cells were accompanied by spike discharges, either facilitated or suppressed by photic stimulation. The cells were also invaded by antidromic impulses, which survived after chemical synapses had been blocked by application of atomized CoC12 solution. In subsequent histology of the Procion-stained neurones, the cell bodies were found in the ganglion cell layer and the axons were often traced.3. Amacrine cells were subdivided into two types. The first type gave rise to transient depolarizations at both on-and offsets of spot and annulus illuminations, usually being associated with spike discharges of which the amplitudes varied in different cells. In histology, the cell bodies of this type were situated in the inner nuclear layer and dendrites ramified in two or more discrete sublayers of the inner plexiform layer (the stratified amacrine cell of Cajal).4. The second type of amacrine cells produced sustained responses during illumination, being associated with no spike but with small oscillatory wavelets. The cell bodies were situated in the inner nuclear layer and the dendrites ramified in a single sublayer of the inner plexiform layer (the monolayered amacrine cell).

Recent studies have shown that cholinergic amacrine cells possess unique membrane properties. However, voltage-gated ionic channels in cholinergic amacrine cells have not been characterized systematically. In this study, using electrophysiological and immunohistochemical techniques, we examined voltage-gated ionic channels in a transgenic mouse line the cholinergic amacrine cells of which were selectively labeled with green fluorescent protein (GFP). Voltage-gated K(+) currents contained a 4-aminopyridine-sensitive current (A current) and a tetraethylammonium-sensitive current (delayed rectifier K(+) current). Voltage-gated Ca(2+) currents contained a omega-conotoxin GVIA-sensitive component (N-type) and a omega-Aga IVA-sensitive component (P/Q-type). Tetrodotoxin-sensitive Na(+) currents and dihydropyridine-sensitive Ca(2+) currents (L-type) were not observed. Immunoreactivity for the Na channel subunit (Pan Nav), the K channel subunits (the A-current subunits [Kv. 3.3 and Kv 3.4]) and the Ca channel subunits (alpha1(A) [P/Q-type], alpha1(B) [N-type] and alpha1(C) [L-type]) was detected in the membrane fraction of the mouse retina by Western blot analysis. Immunoreactivity for the Kv. 3.3, Kv 3.4, alpha1(A) [P/Q-type], and alpha1(B) [N-type] was colocalized with the GFP signals. Immunoreactivity for alpha1(C) [L-type] was not colocalized with the GFP signals. Immunoreactivity for Pan Nav did not exist on the membrane surface of the GFP-positive cells. Our findings indicate that signal propagation in cholinergic amacrine cells is mediated by a combination of two types of voltage-gated K(+) currents (the A current and the delayed rectifier K(+) current) and two types of voltage-gated Ca(2+) currents (the P/Q-type and the N-type) in the mouse retina.

In the isolated, perfused retina of the carp, the effects of GABA and its antagonists, picrotoxin and bicuculline, were investigated on spectral responses of horizontal cells. When the GABA-mediated feedback from horizontal cells to cones was desensitized or blocked by these chemicals, remarkable changes were observed in spectral responses, due to selective suppression of responses to long wavelengths. In particular, the depolarizing responses in the biphasic C-type (H2) cell were abolished, while the hyperpolarizing responses to short wavelengths were retained, resulting in a monophasic spectral response curve. From these results, it was concluded that the GABA-mediated negative feedback plays an essential role in neural mechanisms which convert the trichromatic process at the level of cones into the opponent color process in horizontal cells.

Previous findings have shown that P2X-purinoceptor-mediated signaling pathways regulate the release of ACh in the retina. We previously reported the existence of immunoreactivity for P2X1-, P2X2-, P2X4-, and P2X7-purinoceptors in mouse retina and speculated that P2X2 and P2X7-purinoceptors may modulate the activity of cholinergic amacrine cells. In the present study, we used an immunohistochemical technique to examine whether P2X3-, P2X5, and P2X6-purinoceptors are also important for the modulation of cholinergic amacrine cells in mouse retina. Immunoreactivity for P2X3-, P2X5-, and P2X6-purinoceptors was observed in mouse retina. Immunoreactivity for P2X3- purinoceptors was observed in the dendrites of cholinergic amacrine cells. Immunoreactivity for P2X5-purinoceptors existed in the soma of cholinergic amacrine cells. P2X6-purinoceptor immunoreactivity was not colocalized with the cholinergic amacrine cells. We concluded that, among the three P2X-purinoceptors that were examined, P2X3-purinoceptors seem to affect the function of cholinergic amacrine cells in the mouse retina.