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Distribution of alpha-amino-3-hydroxy-5-methyl-4 isoazolepropionic acid and N-methyl-D-aspartate receptor subunits in the vestibular and spiral ganglia of the mouse during early development.

We investigated the distribution of the glutamate receptor subunits, alpha-amino-3-hydroxy-5-methyl-4 isoazolepropionic acid (AMPA) GluR2 and GluR2/R3, and N-methyl-D-aspartate (NMDA) NR1, and the timing of their appearance during early development of the mouse vestibular and spiral ganglia. NMDA NR1 was the first to be expressed, in the statoacoustic ganglion neurons on E11. GluR2/R3 immunoreactivity was detected in these neurons on E12. This signal probably corresponded exclusively to GluR3, as no signal was obtained for GluR2 alone at this stage. The appearance of these proteins began much earlier than previously reported. GluR2 staining was observed later, on E14 in the vestibular neurons and on E17 in the spiral neurons. The sequence in which these three glutamate receptors appeared suggested possible differences in their roles in the establishment of neuronal circuitry in the inner ear sensory epithelia. The production of NR1 and GluR2/R3 began during the early period of neuron growth and fasciculation. GluR2 appeared later and its expression paralleled synaptogenesis in the vestibular sensory epithelia and in the organ of Corti.

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Regulation of NMDA receptor subunit mRNA expression in the guinea pig vestibular nuclei following unilateral labyrinthectomy.

The localization of neurons expressing mRNAs for the NR1 and NR2A-D subunits of the glutamatergic NMDA receptor was examined by non-radioactive in situ hybridization throughout the guinea pig vestibular nuclei. After deafferentation of the vestibular nuclei by unilateral labyrinthectomy, modifications of the mRNA distributions were followed for 30 days. A quantitative analysis was performed in the medial vestibular nucleus by comparison of the labelled neurons in the ipsi- and contra-lateral nuclei. In vestibular nuclei, the NR1 subunit mRNA was found in various populations of neurons. The NR2A and NR2C subunit mRNAs were less widely distributed, whereas little NR2D mRNA was detected and only rare cells contained NR2B mRNA. NR1 and NR2A-D mRNAs were colocalized in some but not other neuronal types. Twenty hours after the lesion, there was a transient ipsilateral increase of NR1 mRNA level in the medial vestibular nucleus, followed by a decrease 48 h after the lesion and, at 3 days, by recovery to the control level. An ipsilateral increase in the mRNA level of NR2C subunit was detected 20 h after lesion and maintained at 48 h. No significant changes were apparent in NR2A, NR2B and NR2D mRNA levels. The distributions and the differential signal intensities of NR2A-D mRNAs suggest various subunit organizations of the NMDA receptors in different neurons of the vestibular nuclei. Neuronal plasticity reorganizations in the vestibular nuclei following unilateral labyrinthectomy appear to include only changes in NR1 and NR2C mRNA levels modifying the functional diversity of the NMDA receptor in the ipsilateral medial vestibular nucleus neurons. The transient changes in NR1 and the NR2C subunit mRNA expressions in response to sensory deprivation are consistent with an active role for NMDA receptors in the appearance and development of the vestibular compensatory process.

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Heading representation in MST: sensory interactions and population encoding.

Dorsal medial superior temporal cortex (MSTd)'s population response encodes heading direction from optic flow seen during fixation or pursuit. Vestibular responses in these neurons might enhance heading representation during self-movement in light or provide an alternative basis for heading representation during self-movement in darkness. We have compared these hypotheses by recording MSTd neuronal responses to translational self-movement in light and darkness, during fixation and pursuit. Translational movement in darkness, with gaze fixed, evokes transient vestibular responses during acceleration that reverse directionality during deceleration and persist without a fixation target. Movement in light increases the amplitude and duration of these responses so they mimic responses to simulated optic flow presented without translational movement. Pursuit of a stationary landmark during translational movement combines vestibular and visual effects with pursuit responses. Vestibular, visual, and pursuit effects interact so that single neuron heading responses vary across the stimulus period and between stimulus conditions. Combining single neuron responses by population vector summation yields stronger heading estimates in light than in darkness, with gaze fixed or during landmark pursuit. Adding translational movement to robust optic flow stimuli does not augment the population response. Vestibular signals enhance single neuron responses in light and maintain population heading estimation in darkness, potentially extending MSTd's heading representation across the continuum of naturalistic self-movement conditions.

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Postsynaptic inhibition of oculomotor neurons involved in vestibulo-ocular reflexes arising from semicircular canals of rabbits.

In anesthetized albino rabbits, electric stimulation of vestibular nerve branches innervating semicircular canals produced not only reflex contraction in certain extraocular muscles, but also a transient relaxation in others. From relaxing muscles was recorded a slow muscle potential that reflected depression of spontaneous spike discharges in muscle fibers. When recorded monophasically, spontaneous spikes of muscle fibers were superposed to form a direct current potential, and depression of the spikes resulted in a transient reduction of this direct current potential, i.e., the slow muscle potential. The slow muscle potential was correlated to the postsynaptic inhibition induced in oculomotor neurons through the vestibulo-ocular reflex are for the following reasons; its latency was compatible with that of the IPSP's recorded from oculomotor neurons; it was removed by severing axons of the inhibitory second-order vestibular neurons; it was blocked by intravenous injection of picrotoxin as were the IPSP's in oculomotor neurons. By recording slow muscle potentials, a specific canal-muscle relationship for the vestibulo-ocular reflex inhibition of oculomotor neurons was shown to be complementary to that obtained for the vestibulo-ocular reflex excitation.

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Evidence of a collateralized climbing fiber projection from the inferior olive to the flocculus and vestibular nuclei in rabbits.

In albino rabbits, horseradish peroxidase injections confined to vestibular nuclei retrogradely labeled neurons in the dorsal cap of the contralateral inferior olive. Mapping with stimulating electrodes revealed that stimulation of the lateral aspect of the medial vestibular nucleus and the medial aspect of the inferior vestibular nucleus evoked field potentials representing antidromic activation of contralateral dorsal cap neurons. These responses interfered with the antidromic response evoked from the contralateral flocculus and orthodromic responses evoked from the contralateral retina, suggesting that dorsal cap neurons which both receive retinal input and project to the flocculus send collaterals to vestibular nuclei.

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Tilt responses of neurons in the caudal descending nucleus of the decerebrate cat: influence of the caudal cerebellar vermis and of neck receptors.

1. In decerebrate cats with intact cerebellums, we studied the responses of neurons in the caudal areas of the vestibular nuclei to natural vestibular stimulation in vertical planes and to neck rotation. The activity of most neurons was recorded in the caudal half of the descending nucleus. 2. One goal of our experiments was to compare the dynamic and spatial properties of responses to sinusoidal vestibular stimulation with those seen in previous experiments in which the caudal cerebellar vermis, including the nodulus and uvula, was removed. This part of the cerebellum receives vestibular input and projects to the caudal areas of the vestibular nuclei, suggesting that it could influence responses to stimulation of the labyrinth. 3. As in our previous experiments, most neurons could be classified as receiving predominant input either from the otoliths or from one vertical semicircular canal. When mean gain and phase and response vector orientations were compared, there were no obvious differences between the behavior of neurons in the partially decerebellate preparation and the one with the cerebellum intact, demonstrating that in the decerebrate cat the nodulus and uvula have little or no influence on the processing of vertical vestibular input in this region of the vestibular nuclei. 4. Only 23 of 74 (31%) of neurons tested responded to neck rotation. This contrasts with the much larger fractions that respond to this stimulus in Deiters' nucleus and in the rostral descending nucleus. We also recorded from neurons near the vestibular nuclei, mainly in the external cuneate nucleus. All of them (9 of 9) responded to neck rotation. 5. Responses to neck rotation also differed in their dynamics from those found more rostrally in the vestibular nuclei. Dynamics of more rostral neurons resemble those of neck muscle spindles, as do those of external cuneate neurons. The dynamics of caudal vestibular neurons, on the other hand, have a steeper gain slope and more advanced phases than do those of neurons in the more rostral vestibular nuclei. This suggests the possibility of involvement of additional receptors in the production of these responses. 6. In the more rostral vestibular nuclei, responses to vestibular and neck rotation are most often antagonistic, so that head rotation results in little or no response. This is not the case in the caudal areas of the vestibular nuclei, where less than half the neurons tested displayed antagonistic behavior. Further experiments are required to put the neck projection to the caudal vestibular nuclei in a functional context.

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Orienting-related eye-neck neurons of the medial ponto-bulbar reticular formation do not participate in horizontal canal-dependent vestibular reflexes of alert cats.

Ponto-bulbar reticular formation neurons, including identified reticulospinal neurons, were studied in alert, head-fixed cats. Orienting-related neurons of "eye-neck" type (ENNs) were selected on the basis of qualitative correlations of their discharges with visually triggered eye saccades and electromyographic activity (EMG) of dorsal neck muscles. It was tested whether ENNs participate both in visually triggered gaze shifts requiring eye-head coordination and in gaze-stabilizing movements, such as vestibulo-ocular and vestibulo-collic reflexes (VOR, VCR). Firing patterns were studied during passive sinusoidal rotation (0.2-1.0 Hz; 2.0-21.5 deg peak-to-peak) in the horizontal plane. Responses to electrical stimulation of the superior colliculus and the vestibular nerve were recorded to assess the convergence of tectal and vestibular synaptic inputs. The same methods were applied to a control sample of neurons with discharges apparently "unrelated" to orienting movements. ENNs did not show any modulation of firing rate correlated to compensatory VOR or VCR during passive sinusoidal rotations. Among "unrelated" cells, the fraction of modulated units was close to that reported for reticular neurons projecting in the medial reticulospinal tract. Phasic and sustained components of ENN bursts were associated with anticompensatory movements induced by rotation, such as quick phases, ocular beating field shift, and the increase of EMG activity in neck muscles acting in the direction of passive rotation. Monosynaptic excitation from the contralateral superior colliculus was observed in 92.3% of ENNs, but only 2 out of 17 tested showed an excitatory response to vestibular nerve stimulation. In the control group of "unrelated" neurons the proportions of monosynaptic tectal and excitatory vestibular nerve inputs were, respectively, 75.6 and 71.4%. It is concluded that ENNs are specifically related to active gaze shifts, derived from either visual or from head velocity inputs. Rhombencephalic connections of vestibular nuclei to these neurons appear to be quite weak. Parallel inputs from the mid- or forebrain must be assumed to explain their firing patterns during rotation-induced anticompensatory gaze shifts. Within the studied range of frequencies and amplitudes of passive rotation, ENNs did not participate in the vestibulo-collic reflex. It is therefore unlikely that reticular neurons controlling orienting eye-neck synergies act also as a premotor pathway for gaze-stabilizing movements.

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[The effect of vibration on the impulse activity of cortical neurons and their reaction to stimulation of the posterior hypothalamus and Deiters' vestibular nucleus].

The effect of vibration on spontaneous unit activity, oxygen consumption in cortical neurons and their responses to stimulation of posterior hypothalamus and vestibular Deiters' nucleus, were studied. Biphasic changes occurred in frequency of cortical unit activity and in the latter interrelationships with the level of oxygen consumption after 3-hour exposure to the vibration. The 15-day vibration enhanced the mean frequency of unit activity, facilitated the effect of subcortical neurons and eliminated bursts of discharges. The effect of long-term vibration upon cortical unit activity response to stimulation of subcortical structures was quite obvious.

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Secondary vestibular cholinergic projection to the cerebellum of rabbit and rat as revealed by choline acetyltransferase immunohistochemistry, retrograde and orthograde tracers.

Previously we have shown that four regions of the cerebellum, the uvula-nodulus, flocculus, ventral paraflocculus, and anterior lobe 1, receive extensive, but not exclusive, cholinergic mossy fiber projections. In the present experiment we have studied the origin of three of these projections in the rat and rabbit (uvula-nodulus, flocculus, ventral paraflocculus), using choline acetyltransferase (ChAT) immunohistochemistry in combination with a double label, retrogradely transported horseradish peroxidase (HRP). We have demonstrated that in both the rat and rabbit the caudal medial vestibular nucleus (MVN) and to a lesser extent the nucleus prepositus hypoglossus (NPH) contain ChAT-positive neurons. Neurons of the caudal MVN are double-labeled following HRP injections into the uvula-nodulus. HRP injections into the uvula-nodulus also labeled less than 5% of the neurons in the cholinergic vestibular efferent complex. Fewer ChAT-positive neurons in the MVN and some ChAT-positive neurons in the NPH are double-labeled following HRP injections into the flocculus. Almost no ChAT-positive neurons in the MVN and some ChAT-positive neurons in the NPH are double-labeled following HRP injections into the ventral paraflocculus. Injections of Phaseolus leucoagglutinin (PHA-L) into the caudal MVN of both the rat and rabbit demonstrated projection patterns to the uvula-nodulus and flocculus that were qualitatively similar to those observed using ChAT immunohistochemistry. We conclude that the cholinergic mossy fiber pathway to the cerebellum in general and the uvula-nodulus in particular is likely to mediate secondary vestibular information related to postural adjustments.

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Role of visuo-vestibular interaction in pathological ocular oscillation: new model and control system analysis.

Control system engineering is a relatively newer field of engineering which is directly applicable to performance analysis of neuronal networks. Visuo-vestibular interaction (VVI) represents a neuronal control system which has most benefited from such analysis. In this paper, we present a new model of VVI derived from our previously reported model by adding cortical pursuit feedback. Mathematical analysis of abnormal eye oscillations and stability analysis of the VVI system provided us with a plausible hypothesis that the pathological ocular oscillations seen in oculopalatal myoclonus and see-saw nystagmus are indeed due to an unstable VVI control system. The former is due to degeneration of the inferior olivary nucleus resulting in loss of cerebellar learning effects, while the latter is due to disruption of the pathway conveying retinal error signals.

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Vestibular control of swimming in lamprey. I. Responses of reticulospinal neurons to roll and pitch.

A method has been developed for recording the response of single neurons in the lamprey brainstem in vitro to natural stimulation of vestibular receptors. The brainstem dissected together with the intact vestibular apparatus could be rotated in space, in two perpendicular planes (transverse, the roll tilt, and sagittal, the pitch tilt), in one of them up to 360 degrees, and in the other one up to +/- 30 degrees. The responses of single reticulospinal (RS) neurons, in all four reticular nuclei of the brainstem, to roll and pitch were recorded extracellularly and, with small inclinations (up to +/- 45 degrees) also intracellularly. Two types of preparations were used, with and without the rostral part of the spinal cord. In the brainstem preparations, most RS neurons responded both to a definite brain orientation in space and to a change of the orientation (static and dynamic reactions). Responses to roll tilt were similar in all reticular nuclei: all cells were excited with roll tilt towards the contralateral side, this reaction was qualitatively preserved when the roll was performed in combination with different pitch inclinations. Responses to pitch tilt were less clearcut; some neurons were activated with nose-up deflection while others responded to nose-down tilt. In preparations including the spinal cord, responses of RS neurons to roll and pitch tilt differed from those in the isolated brainstem in that they were much less specific and stable. Roll and pitch tilts could trigger the spinal locomotor CPG, which, by sending "efference copy" signals back to the brainstem, produced modulation of RS neurons in relation to the locomotor rhythm.

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Behavior of eye-movement-related cells in the vestibular nuclei during combined rotational and translational stimuli.

1. Secondary position-vestibular-pause (PVP) neurons in the vestibuloocular reflex (VOR) pathway of adult rhesus monkeys were studied during combined semicircular canal and otolith stimulation. The head was rotated at 0.5 Hz with the axis of rotation centered between the otolith organs (on-axis, ON) and with the axis of rotation 23 cm in front of the otoliths (off-axis, OFF). Both conditions were tested with two different vergence angles by the use of 14-cm (near target, NT) and 100-cm (far target, FT) targets. 2. The tangential translational stimulus to the otoliths in the OFF trials should result in a compensatory eye movement that is opposite in direction to that resulting from the angular stimulus to the canals. The otolith stimulus should be great enough to reverse the eye movement response in the NT OFF trials according to geometric calculations. This reversal in eye movement direction occurred as expected although the latency of the reversal (70 ms) was somewhat greater than expected and the magnitude of the reversal was less than predicted solely on the basis of geometric considerations. 3. The responses of the PVP neurons were corrected for eye position sensitivity to investigate the head movement response components. The amplitude of the response in 22 of 24 PVP cells was reduced in the NT OFF condition compared with the FT OFF condition. This difference was not sufficient in itself to explain the observed reversal in eye movement response. 4. The average sensitivities of the neurons to rotation during the FT and NT ON trials were 1.38 and 1.41 spikes.s-1.deg-1.s-1, respectively. This is too small an increase to account for the increase in the angular VOR gain with near targets (approximately 25%); therefore cells other than PVP neurons must be responsible. 5. The average sensitivities of the PVP neurons to translational accelerations obtained from the FT and NT OFF trials were 305 and 484 spikes.s-1.g-1, respectively, which is higher than most otolith afferent sensitivities reported for 0.5-Hz stimuli in the literature. The otolith component is modified by ocular convergence (59% increase in sensitivity), but this increase is too small to account for the change in the translational VOR gain between the two conditions. 6. Although recordings were only obtained from seven eye-head-velocity cells, the results indicate that these neurons may provide the additional signals not present in the PVP cells. These neurons exhibited large differences between ON and OFF rotations and were found to substantially increase their modulation during the NT conditions compared with that observed during the FT conditions.

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An electrophysiological study of pathways mediating optokinetic responses to the vestibular nucleus in the rat.

Intra- and extracellular responses of neurons in the pretectum (Pt), the nucleus reticularis tegmenti pontis (NRTP), the prepositus hypoglossal complex (NPH) and the vestibular nuclei (VN) were recorded during orthodromic/antidromic stimulation of their afferent/efferent fibers. In the Pt, many neurons were excited by stimulation of the contralateral optic nerve (ONc). Comparison of the latencies of evoked presynaptic action potentials and EPSPs yielded a time difference corresponding to one synaptic delay. Forty five per cent of these monosynaptically driven neurons were also excited antidromically from the ipsilateral NRTP. In the NRTP, ONc and Pt stimulations evoked disynaptic and monosynaptic EPSPs, respectively. Thirty six per cent of NRTP neurons orthodromically driven from ONc and/or ipsilateral Pt stimulation were also antidromically invaded from either the contralateral (67%) or the ipsilateral (33%) flocculus but never from both. In the NPH, both ipsilateral Pt and NRTP stimulations excited type II neurons monosynaptically. In addition, EPSPs evoked by Pt stimulation could be mediated to the NPH via a disynaptic route involving the NRTP. In the VN, type II neurons were excited by ipsilateral Pt stimulation. When comparing the latencies of action potentials and EPSPs evoked by Pt stimulation in the NPH and in VN type II neurons respectively, a short, possibly monosynaptic connection, may be postulated between the NPH and the VN. Our results suggest that vestibular neurons may be optokinetically driven from the contralateral eye both via Pt-NPH connections and Pt-NRTP-NPH paths. They also confirm the existence of a transcerebellar route from the Pt via the NRTP to the ipsior contralateral flocculi.

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Properties of cerebellar fastigial neurons during translation, rotation, and eye movements.

The most medial of the deep cerebellar nuclei, the fastigial nucleus (FN), receives sensory vestibular information and direct inhibition from the cerebellar vermis. We investigated the signal processing in the primate FN by recording single-unit activities during translational motion, rotational motion, and eye movements. Firing rate modulation during horizontal plane translation in the absence of eye movements was observed in all non-eye-movement-sensitive cells and 26% of the pursuit eye-movement-sensitive neurons in the caudal FN. Many non-eye-movement-sensitive cells recorded in the rostral FN of three fascicularis monkeys exhibited convergence of signals from both the otolith organs and the semicircular canals. At low frequencies of translation, the majority of these rostral FN cells changed their firing rates in phase with head velocity rather than linear acceleration. As frequency increased, FN vestibular neurons exhibited a wide range of response dynamics with most cells being characterized by increasing phase leads as a function of frequency. Unlike cells in the vestibular nuclei, none of the rostral FN cells responded to rotational motion alone, without simultaneously exhibiting sensitivity to translational motion. Modulation during earth-horizontal axis rotation was observed in more than half (77%) of the neurons, although with smaller gains than during translation. In contrast, only 47% of the cells changed their firing rates during earth-vertical axis rotations in the absence of a dynamic linear acceleration stimulus. These response properties suggest that the rostral FN represents a main processing center of otolith-driven information for inertial motion detection and spatial orientation.

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[Somatotopic organization of the vestibulospinal tract in the toad].

The origin of the vestibulospinal projection in the toad has been investigated by using the method of the retrograde axonal transport of HRP injected at various levels of the spinal cord. The vestibulospinal projection, in this species, was found to be somatotopically organized, since neurons projecting to the cervical segments of the spinal cord were located within the rostromedial part of the ventral vestibular nucleus and those neurons projecting to the lumbosacral segments of the spinal cord were located within the caudolateral part of that nucleus. This pattern of organization of the vestibulospinal projection in amphibia is similar to that described in mammals and birds.

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[The influence of labyrinthectomy on the expression of NMDA R1 mRNA].

OBJECTIVE: To study the effect of labyrinthectomy on the expression of NMDAR1 (N-methyl-D-aspartate receptor subunit R1). METHOD: The in situ hybridization was used to observe the expression of NMDA R1mRNA post-unilateral labyrinthectomy and investigate its effect on vestibular compensation. RESULT: 20 h post-unilateral labyrinthectomy, the NMDA R1mRNA positive neurons in the medial vestibular nucleus increased significantly compared with the unlesioned side(P < 0.01). 40 h-6 d, the positive cells decreased gradually. 6 d post lesion, the positive neuron counts of the lesioned side had no significant difference compared with those of the un-lesioned side (P > 0.05). CONCLUSION: The changing trend of the expression of NMDA R1mRNA in the medial vestibular nucleus is related to the recovery of the vestibular static symptoms after unilateral labyrinthectomy. It is suggested that the modulation of the efficacy of the NMDA R1 play an important role in the compensation of the static symptoms.

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Intervestibular integration and its clinical consequences.

Research during the last twenty years has led to the conclusion that it is time for a more integrated and dynamic view on the physiological and pathophysiological process during clinical vestibular stimulation and perhaps also to reevaluate the interpretation of the findings. Ten patients with absence of function in one labyrinth were exposed to caloric and rotatory stimulation several times during the compensating period of 8 weeks. In the acute state, caloric irrigation of the normal ear with 30 degrees or rotatory stimulation toward the damaged ear resulted in total inhibition of the spontaneous nystagmus. In the chronic state the same investigation reversed the spontaneous nystagmus in the opposite direction. The investigation has shown that it is possible clinically to study the reactions of every single neuron in the vestibular ocular reflex pathway and in this way establish a more topographic diagnosis.

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