On cholinergic mechanisms in the optokinetic nystagmus of the frog: antagonistic effects of muscarinic and nicotinic systems.

In a monocular situation, an intravitreal injection of acetylcholine (ACh) agonists (especially muscarinic agonists like muscarine or oxotremorine) provoked both the suppression of the optokinetic nystagmus (OKN) related to the injected eye, and the appearance of a nasal-temporal (N-T) component in the OKN triggered by the contralateral non-injected eye. These two effects were added in a binocular condition. Similar results were obtained with ACh nicotinic antagonists (D-tubocurarine, alpha-bungarotoxin, hexamethonium and gallamine). ACh muscarinic antagonists (atropine, scopolamine) had only moderate effects and failed to provoke an N-T component in the contralateral OKN. These data show that ACh mechanisms are involved in the control of the frog OKN, especially in directional asymmetry, nicotinic and muscarinic systems acting in an antagonistic way.

Anisotropic inhibition in the receptive field surround of the frog retinal ganglion cells, evidenced by bicuculline and SR 95103, a new GABA antagonist.

When GABA antagonists (picrotoxin, bicuculline methiodide and SR 95103) were intravitreally injected in the frog, they increased the number of spikes of transient retinal ganglion cells, as well as the duration of the response. Thus, the transient pattern of the response became more sustained. GABA antagonists also provoked a marked increase in the size of the receptive field, which might be due to the abolition of the inhibition exerted by the surround upon the centre of the field. In fact, a stimulus applied to the surround of the field simultaneously with one applied to the centre no longer provoked the reduction of the field area nor that of the number of spikes. These are effects which were always observed before drug injection. After picrotoxin injection, the enlarged field was concentric with the initial one, both angular diameters doubled, whereas after bicuculline or SR 95103, the enlarged field was not concentric with the initial one and only one diameter increased. Thus, GABA inhibition appears to be distributed according to an anisotropic spatial pattern. Whether this anisotropy might be an input for direction selectivity in the frog visual system is a topic of discussion. With respect to SR 95103, this compound proved to act like a selective GABA antagonist with long lasting effects.

Possible involvement of cholinergic and glycinergic amacrine cells in the inhibition exerted by the ON retinal channel on the OFF retinal channel.

In the frog retina, the inhibition exerted by the ON channel on the OFF channel was evidenced by the increase in transient ganglion cell OFF responses, when the ON channel was blocked by 2-amino-4-phosphonobutyrate (APB). Intraocular administration of the neurotoxic choline analog ethylcholine mustard arizidinium ion (ECMA) also provoked an increase in the number of spikes of transient ganglion cell OFF responses, without suppressing the ON responses. APB, when administrated after ECMA, abolished the ON responses, but did not modify the OFF responses already increased by ECMA. Neurons located in the inner part of the inner nuclear layer were histologically altered by the toxin, and choline acetyltransferase activity was significantly depressed in ECMA-treated retinas. A double immunostaining experiment showed that amacrine cells containing glycine bear muscarinic binding sites. These results confirm the participation of cholinergic neurons in the inhibition exerted by the ON retinal channel on the OFF retinal channel, and suggest the involvement of a cholinergic/glycinergic loop of amacrine cells in this mechanism.

Glutamatergic separation of ON and OFF retinal channels: possible modulation by glycine and acetylcholine.

When intravitreally injected into the frog in vivo, 2-amino-4-phosphonobutyrate (APB) and cis-2,3-piperidine dicarboxylic acid (PDA) showed opposite effects on ON and OFF retinal channels: APB abolished the ON responses in the electroretinogram and in ganglion cell activity, and increased OFF responses. At the same time the receptive field area was enlarged, and the inhibition exerted by the surround was suppressed. A cholinergic/glycinergic loop involving amacrine cells was suggested to be the pathway of the inhibitory ON input upon the OFF channel. PDA abolished the OFF responses in the ERG and in ganglion cell activity, while increasing the ON response in the ERG and decreasing the ganglion cell sensitivity at ON. The receptive field area was not modified, but the inhibition exerted by the surround was suppressed, probably by a blockade of horizontal cell glutamate receptors.

On GABAergic mechanisms in the optokinetic nystagmus of the frog: effects of bicuculline, allylglycine and SR 95103, a new GABA antagonist.

In a monocular situation, an intravitreal injection of the GABA antagonists, bicuculline or SR 95103 provoked both the suppression of the optokinetic nystagmus (OKN) related to the injected eye and the appearance of a Nasal-Temporal (N-T) component in the OKN triggered by the contralateral non-injected eye (this N-T component being absent in control OKN). These two effects were added in a binocular condition. Similar results were obtained with L-C allylglycine which reduces the endogenous GABA level, but these effects were delayed when compared to those of GABA antagonists. All these data are roughly analogous to those previously obtained with picrotoxin (a non-competitive GABA antagonist) and thus confirm that GABA mechanisms are involved in the control of the frog OKN. Furthermore, SR 95103 acted in this model as a potent selective GABA antagonist, as has been demonstrated in another system.

N-methyl-D-aspartate antagonists suppress the development of frog symmetric monocular optokynetic nystagmus observed after unilateral visual deprivation.

In monocular vision, frogs display a unidirectional optokinetic nystagmus (OKN), reacting only to temporal-nasal (T-N) stimulation. The OKN N-T component is almost absent. However, prolonged monocular visual deprivation by unilateral eyelid suture provoked the appearance of the N-T component. The analysis of search coil recordings showed that the slow phase velocity gain of both T-N and N-T components became similar. Chronic administration of N-methyl-D-aspartate (NMDA) antagonists for the duration of deprivation prevented the appearance of a symmetrical monocular OKN in frogs: following repeated intraperitoneal injections of either MK 801, CGS 19755 or intrapretectal microinjections of 2-amino-5-phosphonovalerate (APV), the N-T component did not appear, and OKN remained asymmetrical. Thus NMDA receptors appear to be involved in the control of the plasticity process which allows monocular OKN of adult lower vertebrates to become symmetrical.

Abolition of monocular optokinetic nystagmus directional asymmetry after unilateral visual deprivation in adult vertebrates: involvement of the GABAergic mechanism.

In lower vertebrates such as frogs and chickens, monocular optokinetic nystagmus (OKN) displays directional asymmetry, temporal-nasal (T-N) stimulation being more efficient in evoking this visuomotor reflex than N-T stimulation. The N-T component of monocular OKN is significantly weaker in chickens, while it is almost absent in frogs. Coil recordings showed that in adult frogs and chickens, prolonged monocular visual deprivation by unilateral eyelid suture provoked the appearance of the N-T component in frogs as well as its significant and progressive increase in both species. The administration of THIP, a GABAA agonist, abolished reversibly the increase of the N-T component in both species. This fact suggests that the GABAergic system could be involved in determining this plasticity process observed in adult lower vertebrates.

Plasticity of the frog monocular OKN: involvement of pretectal GABAergic and cholinergic systems.

The frog horizontal monocular optokinetic nystagmus (OKN) is asymmetrical, the temporal-nasal (T-N) stimulation being the sole stimulation efficient to evoke the reflex, the nasal-temporal (N-T) component being almost absent. Coil recordings showed that, in adult animals, prolonged monocular visual deprivation by unilateral eyelid suture provoked the appearance of the N-T component. The OKN became symmetrical, reacting for both directions of stimulation. Microinjection of either gamma-aminobutyric acid (GABAA) agonist 4,5,6,7-tetrahydroisoxazolo (5,4-C) Pyridin-3-ol (THIP) or muscarinic cholinergic antagonist atropine into the nucleus lentiformis mesencephali, the pretectal mesencephalic structure involved in OKN, transiently abolished the presence of N-T component. This result suggests that the phenomenon of visual plasticity, occurring after a week of monocular deprivation, can be due, at least partially, to reduction in pretectal GABAergic inhibition, and to concomitant activation of cholinergic muscarinic receptors.

Is a retinal input involved in the generation of eye resetting fast phases in the frog eye optokinetic nystagmus?

An intravitreal injection of cis-2,3-piperidine dicarboxylic acid (PDA), a glutamate analog, in one eye only, decreased or even totally suppressed the eye resetting fast phases (ERFPs) of the frog optokinetic nystagmus (OKN) in monocular as in binocular situations. On the opposite, for low drum speeds, the slow phase eye velocity was not affected by PDA. Moreover, it seems that intravitreally injected PDA does not act upon central structures responsible for OKN. Our experiments suggest that a retinal input may be involved in triggering the ERFPs in the OKN.

Are retinal or mesencephalic dopaminergic systems involved in monocular optokinetic nystagmus asymmetry in frog?

In monocular vision, frogs display a unidirectional optokinetic horizontal nystagmus (H-OKN) reacting only to temporal-nasal (T-N) stimulation. The N-T component is almost absent. The analysis of search coil recordings after administration of dopamine into the viewing eye, the occluded eye or directly into the pretectum, hardly modifies the H-OKN triggered by the viewing eye irrespective of the concentration used. Conversely, administration of Piribedil, a strong D2 dopamine agonist, provokes the appearance of a N-T component, suppressing the monocular H-OKN asymmetry, whether the drug is injected by intravitreal or intrapretectal route. It is suggested that Piribedil could also bind with receptors other than dopamine's.

Different effects of dopamine and piribedil (a dopamine D2 agonist) on frog monocular optokinetic nystagmus asymmetry.

Frog monocular optokinetic nystagmus (OKN) displays a directional asymmetry, reacting only to stimulations in the temporal-nasal (T-N) direction. The nasal-temporal (N-T) component is almost absent. The systemic or intrapretectal injection of Piribedil, a D2 dopamine agonist, provokes the appearance of a N-T component suppressing the monocular OKN asymmetry. Conversely, dopamine or haloperidol (a dopamine antagonist, acting mainly on D2 receptors) have no effect upon the monocular OKN unidirectionality. The monocular OKN N-T component still appears after administration of Piribedil even if this injection is preceded by administration of haloperidol which blocks the dopaminergic D2 receptors. Moreover administration of atropine (a cholinergic muscarinic antagonist) following that of Piribedil suppresses the N-T component; when injected before Piribedil, atropine prevents the appearance of the N-T component. These results suggest that in our experiments, Piribedil binds with muscarinic receptors.

Involvement of NMDA in a plasticity phenomenon observed in the adult frog monocular optokinetic nystagmus.

The frog horizontal monocular optokinetic nystagmus (H-OKN) is asymmetrical, the reflex being evoked by a temporal-nasal (T-N) component, but not by a nasal-temporal (N-T) component. Coil recordings showed that, in adult animals, 8 days of monocular deprivation (by unilateral eyelid suture) provoked the appearance of a N-T component, the H-OKN becoming symmetrical, reacting for both directions of stimulation. This delay was shortened to 2 days following two successive unilateral pretectal administrations of NMDA or of LY 285 265, an NMDA agonist, the first 2 days of eyelid suture. The same results were obtained when chronic microinjections of NMDA or LY 285 265 were achieved, the frogs being maintained in total darkness during the week of eyelid suture. These data indicate that the plasticity phenomenon evidenced in the monocular frog H-OKN depends on the activation of the NMDA receptors of one pretectum. This activation was obtained either by a monocular light stimulation of 8 days duration, or by unilateral administration of drugs activating the NMDA glutamatergic pretectal system. In this last case, the light stimulation was no longer necessary.

Physiological effects of muscarinic vs nicotinic ACh antagonists upon ganglion cell activity in the frog retina.

The intravitreal administration of ACh agonists (eserine, carbachol, oxotremorine) or that of ACh muscarinic antagonists (scopolamine, atropine) provoked a reduction of the On-Off ganglion cell discharges. The agonists depressed the Off discharges more than the On discharges, while the ACh muscarinic antagonists depressed the On- more than the Off discharges. These drugs did not modify the ganglion cell receptive field area; thus, the muscarinic cholinergic system seems not to be involved in the spatial organization of the On-Off ganglion cells, but rather seems to play an important part in the separation of On and Off information channels. ACh nicotinic antagonists [hexamethonium, D-tubocurarine (D-TC), alpha-bungarotoxin (alpha-BGTX)] provoked an increase of the receptive field area of On-Off ganglion cells, this enlargement being due to the suppression of the inhibition normally exerted by the surround upon the centre of the field. Moreover D-TC and alpha-BGTX, but but hexamethonium, increased the number of ganglion cell discharges. These data are analogous to those obtained after administration of GABA antagonists and show that through nicotinic receptors, ACh seems to be involved in the spatial organization of the On-Off ganglion cell.

Directional asymmetry of the frog monocular optokinetic nystagmus: cholinergic modulation.

The frog monocular optokinetic gaze nystagmus (OKN) was studied by coil recordings after intravitreal administration of cholinergic drugs into the closed eye. Before injection, the frog displayed OKN for stimulations in the temporo-nasal (T-N) direction only. The injection of muscarinic agonists, as well as that of nicotinic antagonists, provoked the appearance of a naso-temporal (N-T) component, the slow phase velocity gain then being strongly and significantly increased. The abolition of the OKN directional asymmetry indicates that acetylcholine seems to act in opposite ways through muscarinic and nicotinic binding sites. The GABAergic and cholinergic systems may interact to generate and modulate OKN in the frog.

Pharmacological study of the chicken's monocular optokinetic nystagmus: involvement of the ON retinal channel evidenced by the glutamatergic separation of ON and OFF pathways.

The chicken's monocular optokinetic nystagmus (OKN), recorded by the magnetic search coil technique, displays a directional asymmetry, the temporal-nasal (T-N) stimulation being more efficient than the nasal-temporal (N-T) one to evoke the reflex. The intravitreal administration of APB, a glutamate agonist which selectively blocks the ON retinal channel strongly reduced the eye monocular OKN; it also induced spontaneous eye movements in the T-N direction. The intravitreal injection of PDA another glutamate analog, which reduces the OFF channel, while increasing the activity of the ON channel, induced a large increase in OKN velocity gain, especially for a N-T stimulation at the lowest drum speeds. These results indicate the main involvement of the ON retinal channel in the OKN genesis and the inhibitory effect of the OFF channel upon this oculomotor reflex.

Pharmacological study of the chicken's monocular optokinetic nystagmus: effects of GABAergic agonist and antagonists.

When injected into the chicken open eye, the GABA-agonist THIP and the GABA-antagonists bicuculline and picrotoxin induced spontaneous eye movements in nasal-temporal (N-T) and in temporal-nasal (T-N) direction, respectively. These spontaneous movements were scarcely modulated by optokinetic stimulation, irrespective of the direction of stimulation. It is suggested that they are due to the suppression of directional selectivity of retinal ganglion cells. When injected into the closed eye, GABAergic drugs did not produce spontaneous nystagmus. THIP provoked a reduction of the N-T component, without modifying the T-N one, while GABA antagonists induced a significant increase in OKN performance, especially for the N-T direction of stimulation. In these conditions, picrotoxin also provoked an increase in the duration of both components of optokinetic after nystagmus, indicating a direct effect of the drug upon the velocity-storage system.

Directional asymmetry of the horizontal monocular head and eye optokinetic nystagmus: effects of picrotoxin.

Frog monocular eye and head optokinetic nystagmus (OKN) were studied by coil recordings after intravitreal administration of picrotoxin into the closed eye. Before injection, the frog displayed an OKN only for stimulations in the temporo-nasal (T-N) direction. The injection of picrotoxin provoked the appearance of a N-T component of the head and eye OKN: the slow phase velocity gain and the resetting fast phase frequency were strongly and significantly increased. Thus, picrotoxin abolished the directional asymmetry of head and eye OKN, indicating the involvement of GABAergic mechanisms in the inhibition of the N-T component of the monocular eye and head OKN. Picrotoxin administration had an additional effect on the monocular head OKN only, the performances (measured by the velocity gain and the frequency of resetting fast phases) were markedly increased for both directions of stimulation, suggesting an effect of the drug upon the motor output of head movements.

The pretectal cholinergic system is involved through two opposite ways in frog monocular OKN asymmetry.

Frog monocular horizontal optokinetic nystagmus (OKN) has been studied by coil recordings, before and after unilateral microinjection of cholinergic drugs into the pretectum. The recorded eye was either contralateral or ipsilateral to the injected structure. Before injection, monocular OKN displayed a directional asymmetry, reacting only to stimulations in the temporonasal (T-N) direction. The intrapretectal administration of a cholinergic muscarinic agonist (oxotremorine), as well as that of a nicotinic antagonist (D-tubocurarine), abolished the monocular OKN asymmetry, inducing the appearance of the naso-temporal (N-T) component; the difference between the slow phase velocity gain of both components was no longer significant. These data suggest that acetylcholine (ACh), at the level of the pretectum, acts in opposite ways through muscarinic and nicotinic binding sites; monocular OKN asymmetry could result, at least partially, from a facilitating nicotinic effect and an inhibitory muscarinic effect. Possible interactions with other transmitter systems are discussed.

[Interactions between GABAergic and cholinergic mechanisms involved in monocular optokinetic nystagmus in frogs]. / Interactions de mécanismes GABAergiques et cholinergiques impliqués dans le nystagmus optocinétique monoculaire de la grenouille.

The systemic administration of atropine, a muscarinic cholinergic antagonist, was found to suppress the Nasal-Temporal (N-T) component of the frog monocular optokinetic nystagmus (OKN), which had appeared following a prior injection of bicuculline and which does not exist in the normal animal. On the contrary, the administration of a nicotinic cholinergic antagonist (D-TC, alpha-BGT, Hexamethonium) following that of bicuculline has prolonged the duration of the induced N-T component. Thus, ACh was shown to attenuate or to reinforce the GABAergic inhibition of the N-T component through muscarinic receptors or nicotinic receptors respectively. These data point to the existence of strong interactions between these two neurotransmission systems involved in frog monocular OKN.

Unilateral pretectal microinjections of SR 95,531, a GABA A antagonist: effects on directional asymmetry of frog monocular OKN.

Monocular eye movements have been studied in frogs using the search coil technique before and after unilateral microinjection of SR 95,531, a GABA A antagonist, into the pretectal nuclei contralateral to the open eye. Before injection, monocular, horizontal optokinetic nystagmus (OKN) in frogs, as in other lower vertebrates, displays a directional asymmetry: the stimulation in the T-N (temporo-nasal) direction is more efficient in evoking OKN than is stimulation in the N-T (naso-temporal) direction. The N-T component is almost absent and displays only slow phases of very low speed. Unilateral SR 95,531 microinjection into the pretectum reversibly decreased the directional asymmetry of monocular horizontal OKN, by strongly increasing the N-T component slow phase velocity while the T-N slow phase velocity remained unchanged. These data show that SR 95,531 injected into the pretectum contralateral to the open eye reversibly decreased the inhibition upon the N-T component of monocular horizontal OKN, which suggests that a pretectal GABAergic system is involved in the directional asymmetry of monocular horizontal OKN in frogs.