Two Types of Voltage-activated Calcium Currents in Goldfish Horizontal Cells

In horizontal cells (HCs) that were freshly dissociated from goldfish retina, two types of voltage- dependent calcium currents (ICa) were recorded using a patch-clamping configuration: a transient type current and a sustained type current. The cell was held at -40 mV, and the prepulse step of -90 mV was applied before command pulse between -65 and +55 mV. The transient Ca2+ current was activated by depolarization to around -50 mV from a prepulse voltage of -90 mV lasting at least 400 ms and reached a maximal value near -25 mV. On the other hand, the sustained Ca2+ current was induced by pre-inactivation for less than 10 ms duration. Its activation started near -10 mV and peaked at +20 mV. Co2+ (2 mM) suppressed both of these two components, but nifedipine (20microM), L-type Ca2+ channel antagonist, blocked only the sustained current. Based on the activation voltage and the pharmacological specificity, the sustained current appears to be similar to L-type ICa and the transient type to T-type ICa. This study is the first to confirm that transient type ICa together with the sustained one is present in HCs dissociated from goldfish retina.

Effects of Nitric Oxide on Inhibitory Receptors of Rod Bipolar Cells of Rat Retina

The effects of nitric oxide (NO) on inhibitory neurotransmitter receptors and some types of inhibitory receptors in dissociated rod bipolar cell (RBC) were investigated. In the whole cell voltage-clamping mode, the gamma-aminobutyric acid (GABA) activated current showed both sustained and transient components. GABA activated transient current was fully blocked by bicuculine, a GABAA receptor antagonist. The cis-4-aminocrotonic acid (CACA), a GABAC receptor agonist, evoked the sustained current that was not blocked by bicuculline (BIC). Glycine activated the transient current. These results indicate that the RBCs possess GABAA, GABAC, and glycine inhibitory receptors. Sodium nitroprusside (SNP), a NO analogue, reduced the currents activated by GABAA receptor only, however, did not reduce the currents activated by either GABAC or glycine receptors. This study signifies further that only NO depresses the fast inhibitory response activated by GABAA receptor in RBC. We, therefore, postulate that NO might depress the light-on/off transient inhibitory responses in RBCs in the rat retina.

The Effects of N-Methyl_D-Aspartic Acid, and Antagonism by Kynurenic Acid on Neurons in the Cathish Retina

To investigate the mechanism of the excitatory signal transmission, the effects of N-methyl-D-aspartate(NMDA, ionotropic glutamate agonist) and kynurenic acid(glutamate antagonist) on catfish retinal neurons were explored using conventional intracellular recording techniques. Horizontal cells were depolarized by glutamate, kainate, quisqualate, and NMDA but gyperpolarized by kynurenate. Transient components of both ON-and OFF-bipolar cells were reduced either by glutamate or by NMDA. Kynurenate suppressed sustained components of the third-order neurons or OFF-bipolar cells. Furthermore, kynurenate blocked the depolarizing actions of NMDA on horizontal cells and ON-sustained cells with large ON-transient components. The results suggest that NMDA would exert a tonic depolarization in the horizontal cells and the 3rd-order neurons, and there might be a preferential suppression on the a NMDA receptors by kynurenic acid in the catfish retina.

Effects of the Center-Surround Light Stimuli on the Catfish Retianl Neurons

For the investigation of the receptive field of the retinal neurons, the spatial and temporal properties of horizontal cells and the center-surround antagonisms of bipolar cells and the third-order neurons were studied using conventional intracellular recording techniques. Horizontal cells were hyperpolarized by the large annulus light stimuli (id: 3.5mm) and required at least 0.2 seconds of dark periods to respond enough. The amplitudes of the response of the ON-bipolar cell were decreased as the diameter of the spot stimuli was increased to 2.6mm and the responses were inverted at 2.6mm of inner diameter of the spreading annulus stimuli with fixed width. ON-sustained cell with spikes generated ON-sustained light responses by small annulus (1400-2100micrometer), but elicited OFF-sustained responses by large annulus (2100-2800micrometer). ON-sustained cell without spikes also showed surround antagonism. The spikes were generated at the spot of 490micrometer diameter and they were disappeared at the spot of 5600micrometer diameter. And, OFF-transient component of the ON-OFF transient cell was enhanced as annulus stimuli spread. The results suggest that the horizontal cells have large and monotonic receptive fields while the bipolar cells and the ON-sustained cells have large biphasic receptive fields in the catfish retina.

Chemical coupling between horizontal cells in the catfish retina

The effects of GABA and glutamate on the horizontal cells were explored by an intracellular recording method to discern the mechanisms of receptive field formation by chemical coupling in the catfish outer retina. The results suggest that the horizontal cells of the catfish retina might use GABA as their transmitters and that the GABAergic system contributes to the formation of receptive fields of the horizontal cells. GABA|c receptors may be involved in a chemical coupling between horizontal cells and concerned with the depolarizing actions by GABA on horizontal cells in the catfish retina. Since the chloride equilibrium potential is more positive than the dark membrane potential in horizontal cells, GABA released from a horizontal cell may depolarize the neighboring horizontal cells. Thus a chemical coupling between horizontal cells may be formed. GABA|A receptors also may be involved in the negative feedback mechanism between photoreceptor and horizontal cell. And glutamate may be involved in connecting positive and negative feedback systems since it potentiated the GABA's actions. Therefore, it is presumed that large receptive fields in the catfish retina are formed not only by electrical coupling but also by chemical coupling between horizontal cells. And information travels laterally by pathways involving both electrical coupling composed of gap junctions and chemical coupling in the retinal network.

Preferential Suppression of the On Pathway by r-Aminobutyric Acid in the Catfish Retina

The effects of r-aminobutyric acid(GABA) agonsits and antagonists were explored by the intracellular recording method to discern the preferential suppression of the ON component by GABA on the ON-OFF transient cell in the catfish (Ictalurus punctatus) retina. Experiments were performed in the superfused eyecup preparation. The animals were decapitated and pited before the eye, and the surrounding tissue was removed from the skull. The retina was exposed by excising the cornea, iris, and vitreous. This preparation rested on a wad of Ringer`s soaked cotton in contact with an Ag/Agcl reference electrode. Solutions were delivered through a manifold system that was connected to a pipette located near the absorbent wick. Electro-physiological recordings were made using standard intracellular electrodes filled with 2 M potassium acetate. The electrical signal was recorded with an amplifierand a penwriter, viewed on an oscilloscope, and stored on a data recorder. The light sources were red light-emitting-diode (LED) and the stimuli were full field illumination covering the cntire retina. GABA preferentially reduced ON light responses in ON-OFF transient cell. and GABA hyperpolarized bipolar cells, but the effects on ON bipolar cells were more sensitive than OFF bipolar cells. CACA and TACA, GABAc receptor agonist, did not act on bipolar cells. CACA and TACA, GABAc receptor agonists, hyperpolarized bipolar cells but the sensitivity deferences between ON and OFF bipolar cell were not observed. These results suggest that the preferential suppression of the ON component of the ON-OFF transient cell by GABA was resulted from the presynaptic mechanism that reduced bipolar cell input.

Effects of Extracellular Chloride Ions on the Catfish Retinal Neurons

The catfish (Ictalurus punctatus) retinal neurons were investigated by using the intracellular recording techniques to analyze the function of the chloride ions in the light responses and the ionic mechanisms of the depolarizing actions by GABA. Experiments were performed in the superfused retina-eyecup preparation. The retina was exposed by exicising the cornea, iris, and vitreous. A piece of absorbent tissue with a hole large enough to expose the retina was centered over the eyecup to serve as a wick to draw off the superfusate. Diffuse light stimuli were generated by light-emitting diode positioned above the eyecup. The recordings were made with the use of borosilicate glass micropipettes fashioned from' omega dot' capillary tubing filled with 2 M potassium acetate. Voltage recordings were obtained using an amplifier and amplified signals were recorded on a storage oscillocope, penwriter, and a data recorder. In the catfish retina, the dark membrane potentials were depolarized and the light evoked responses were enhanced in the chloride"-free medium on the catfish horizontal cells. The amplitude of the light evoked potentials were increased by chloride free Ringer's solution on the ON- and OFF-bipolar cells. But the dark membrane potentials were hyperpolarized on the ON-bipolar cell and depolarized on the OFF-bipolar cells in the chloride free medium. The chloride free Ringer's solution changed the light response from ON-sustained to OFF-sustained without any change in amplitude on the ON-sustained cell. The depolarizing actions by GABA on the horizontal cells were maintained in chloride-free environment. But GABA did not abolished the light evoked potentials of the horizontal cell and the ON-sustained cell under the chloride free environment. The results suggest that chloride ion has important roles on the signal transmission of the dark periods in the catfish retina and the depolarizing actions by GABA on the neurons in the catfish retina might be chloride dependent.