Polysynaptic inputs to vestibular efferent neurons as revealed by viral transneuronal tracing.

The Bartha strain of the alpha-herpes pseudorabies virus (PrV) was used as a retrograde transneuronal tracer to map synaptic inputs to the vestibular efferent neurons of the Mongolian gerbil, Meriones unguiculatus. Although previous experiments have shown that vestibular efferent neurons respond to visual motion and somatosensory stimuli, the anatomic connections mediating those responses are unknown. PrV was injected unilaterally into the horizontal semicircular canal neuroepithelium of gerbils, where it was taken up by efferent axon terminals. The virus was then retrogradely transported to efferent cell bodies, replicated, and transported into synaptic endings projecting onto the efferent cells. Thirty animals were sacrificed at approximately 5-h increments between 75 and 105 h post-infection after determining that shorter time points had no central infection. Infected cells were visualized immunohistochemically. Temporal progression of neuronal infection was used to determine the nature of primary and higher order projections to the vestibular efferent neurons. Animals sacrificed at 80-94 h post-inoculation exhibited immunostaining in the dorsal and ventral group of vestibular efferent neurons, predominately on the contralateral side. Neurons within the medial, gigantocellular, and lateral reticular formations were among the first cells infected thereafter. At 95 h, additional virus-labeled cell groups included the solitary, area postrema, pontine reticular, prepositus, dorsal raphe, tegmental, the subcoeruleus nuclei, the nucleus of Darkschewitsch, and the inferior olivary beta and ventrolateral subnuclei. Analysis beyond 95 h revealed virus-infected neurons located in the vestibulo-cerebellar and motor cortices. Paraventricular, lateral, and posterior hypothalamic cells, as well as central amygdala cells, were also labeled. Spinal cord tissue exhibited no labeling in the intermediolateral cell column, but scattered cells were found in the central cervical nucleus. The results suggest functional associations among efferent feedback regulation of labyrinthine sensory input and both behavioral and autonomic systems, and support a closed-looped vestibular feedback model with additional open-loop polysynaptic inputs.

Asymmetric gene expression in the brain during acute compensation to unilateral vestibular labyrinthectomy in the Mongolian gerbil.

Commercial microarrays were used to identify transcriptome expression within vestibular related brain regions (vestibular brainstem and cerebellum, and caudotemporal cortical regions) during the acute period of recovery following unilateral surgical vestibular labyrinth ablation in the gerbil. As a representative model of vestibular compensation, vestibular lesions in the gerbil produced activation in a common set of genes related to vestibular compensation. The total RNA was prepared and amplified using Affymetrix Gene Chip probes from the Rat U34 Neurobiology and R230, and Mouse M430 gene sets, resulting in GCRMA summarized data from S+AA software. Matched rat and mouse genes from gerbil hybridization produced good interspecies synteny. Multiple gene target trends supported global increases in neuron excitability throughout the vestibular brainstem and cerebellum. We focused further on gene expression with anatomically asymmetric activation relative to the lesion, indicative of involvement in rebalancing central vestibular tone during the vestibular compensation process. Cluster analysis revealed distinct spatial (regional and ipsi-contra) and temporal patterns. The asymmetric genes were part of well-defined neuron-related networks and included multiple members of the glutamate and GABA neurotransmitter systems. Transcripts for D3 dopamine, glycine, and some GABA receptor signals increased quickly in the ipsilesional vestibular complex and then increased gradually in the contralateral region, restoring the expression symmetry. Alternatively, the NMDA binding subunit decreased gradually over the acute compensation period in the contralateral vestibular complex. There was evidence for numerous associations between signaling systems with PKC as one possible mediator between early changes in GABA and progressive changes in NMDA signaling. These data begin to define the compensatory response at the level of molecular cascades.

Fos expression in the vestibular brainstem: what one marker can tell us about the network.

Fos inducible transcription factor expression in rodent brains (rats and gerbils) during manipulations of vestibular input is reviewed. Stimuli included centripetal hypergravity, unilateral labyrinth lesion or semicircular canal plugging, rotational axis cross-coupling (Coriolis forces), high and low rotational vestibulo-ocular reflex gain adaptation, translabyrinth galvanic stimulation, pharmacological manipulation, and combinations thereof. Each type of stimulation elicited unique but partially redundant response patterns in the vestibulo-olivo-cerebellar (VOC) network that reflect the origin and interaction of the labyrinth inputs. On the basis of these patterns, a trained observer can predict what the animal experienced during testing; the patterns of VOC Fos expression reveal a trace of recent genomic activity. Based on principal component analysis, VOC network modules associated with lesion recovery, spatial representation and the calibration of gravity, and optokinetic influences are proposed. Probable and possible gene targets of the Fos protein are also reviewed.

Fos responses to short-term adaptation of the horizontal vestibuloocular reflex before and after vestibular compensation in the Mongolian gerbil.

Fos expression in vestibular brainstem and cerebellar regions was evaluated during vestibular adaptation in the Mongolian gerbil. In addition, vestibular adaptation was evaluated in both normal and compensated animals, as vestibular compensation reorganizes the vestibular pathway constraining adaptive processes. Behaviorally, discordant optokinetic and vestibular input induced appropriate high and low gain in horizontal angular vestibuloocular reflex responses. In normal animals, low gain adaptation was more complete than high gain. However, in compensated animals, only low gain adaptation produced adaptive responses both toward and away from the lesion with appropriate gain shifts. High gain adaptation in compensated animals failed to result in gain adaptation for head movements toward the side of the lesion. Fos expression during acute vestibular adaptation in normal animals was found in the flocculus/paraflocculus, the dorsal cap of the inferior olive (IOK), and the prepositus hypoglossi (PrH). Floccular Fos labeling was increased under both high and low gain conditions. IOK and PrH labeling was increased and correlated during low gain conditions, but was reduced and uncorrelated during high gain conditions. The pattern of Fos labeling in compensated animals was asymmetric-favoring the ipsilesional flocculus and contralesional vestibular brainstem. Both compensated high and low gain adaptation groups displayed increased floccular and IOK Fos labeling, but only compensated high gain adaptation produced increased Fos labeling in the medial vestibular nucleus. The behavioral and Fos labeling results are consistent with visual-vestibular adaptation requiring direct vestibular input.

VOR and Fos response during acute vestibular compensation in the Mongolian gerbil in darkness and in light.

We measured binocular horizontal eye movements in the gerbil following unilateral labyrinthectomy during the acute phase (1-24 h) of vestibular compensation. Regardless of whether the animals compensated in the light or the dark, VOR gain progressively reduced following the lesion, and normal oculomotor symmetry was disrupted. Initially, the VOR was comparable at 1 h post-lesion for both visual conditions. However, by 3 h post-lesion the VOR response for head turns away from the lesion continued to drop in animals compensating in the dark. By 24 h, both groups displayed reduced VOR gains, but animals compensating in the light had improved frequency response characteristics. Optokinetic responses became unstable but were generally elevated compared to pre-lesion levels. Animals with vision had reduced optokinetic gains by 24 h, while the OKR response for animals in the dark remained elevated. Brainstem Fos labeling generally increased from 1 to 3 h, then decreased by 24 h. However, at 1 h, Fos labeling in the inferior olivary dorsal cap and prepositus contralateral to the lesion was significantly increased in animals compensating in the light. In both visual conditions, flocculus and paraflocculus Purkinje cell labeling was also observed, and some of the Fos-labeled cells in the medial vestibular nucleus were commissural. Fos in the dorsal cap and prepositus could be attributed to the presence of visual input. While the visually related prepositus Fos labeling preceded improved VOR performance, the dorsal cap appeared to be involved in resolving visual and motor deficits from spontaneous nystagmus.

Relationship of static and dynamic mechanisms in vestibuloocular reflex compensation.

OBJECTIVES/HYPOTHESIS: We tested the hypothesis that recovery of dynamic oculomotor behavior (specifically the vestibuloocular reflex [VOR]) after a unilateral labyrinthectomy (UL) is independent from static deficit recovery (e.g., spontaneous nystagmus). This hypothesis predicts that VOR recovery from peripheral vestibular lesions that do not cause static symptoms, such as unilateral semicircular canal plugging (UCP), would have a similar time course and magnitude as recovery from a lesion that creates both a static and dynamic imbalance, such as UL. Furthermore, animals compensated after UCP would be expected to retain their compensated VOR response after the additional insult of ipsilateral labyrinthectomy. STUDY DESIGN: An experimental study in the Mongolian gerbil animal model. METHODS: The horizontal VOR was measured from both eyes using infrared video-oculography on gerbils before and after UCP, UL, or ipsilateral labyrinthectomy after a previous UCP. Eye movements were recorded during yaw rotation in the dark. RESULTS: UL resulted in a more severe acute deficit than the UCP. Over several weeks, the UCP animals compensated their horizontal VOR, particularly on rotation toward the intact side, quicker and more completely than the UL animals. Animals that underwent ipsilateral labyrinthectomy 8 to 11 weeks after UCP demonstrated preservation of the improved gain, particularly on rotation toward the intact labyrinth. However, the difference between the UL groups with or without precedent UCP was not retained after 72 hours, and long-term compensation was poorest in the UL after UCP group. CONCLUSIONS: Plasticity in dynamic vestibular reflexes induced by UCP is preserved after a subsequent UL. However, neurologic events during the first and second day after UL appear to limit, change, or suppress the long-term dynamic compensation of the VOR, regardless of whether the animal had a previous UCP.

Video-oculography in the gerbil.

Normative vestibulo-ocular and optokinetic reflexes (VOR and OKR) and pupil diameter were measured in young adult gerbils using infrared video-oculography with 60 Hz sampling during head-fixed binocular recordings. The pupillary light-sink technique was preferred over a single-beam retinal reflection method because its measurements were less affected by pupil size. Eye movements were generally conjugate with occasional independent saccadic movements, and independent drifting movements in the dark. The horizontal optokinetic response to sinusoidal motion of a randomly spaced white dot pattern was maximal at low velocities (5 degrees/s), stronger temporonasally, and dropped off quickly at approximately 20 degrees/s. Constant velocity gain was near unity through 60-80 degrees/s with a sharp drop-off. Monocular viewing revealed almost no nasotemporal optokinetic response. Pupil diameter was found to vary as a saddle function with optokinetic gain from cycle to cycle, but also have a circadian rhythm (smaller at dusk) that related inversely to mean horizontal VOR gain. Gerbils with eyes open sometimes had no optokinetic response during long stimulus periods, which then resumed after a brief vestibular stimulus. The horizontal angular VOR gain was relatively flat across 0.1-1.0 Hz and 30-120 d/s sines (phase near zero), with a mean gain of approximately 0.78 in the dark, and 1.0 with the fixed pattern surround (n=15, for both raw calibrated and normalized data). Most animals also revealed a strong slow phase eye velocity asymmetry (dominant during ipsilateral rotation) in the half-cycle gain of their horizontal angular VOR response in the dark. A constant velocity horizontal optokinetic bias velocity did not change the gain or symmetry of the sinusoidal VOR response, but shifted the VOR response velocity in an additive (linear) fashion. Both cross-coupling (pitch or roll while rotating) and pseudo-OVAR (off-axis counter-rotation) stimuli generated horizontal nystagmus. The findings suggest that the gerbil, like other lateral-eyed rodents, relies on otolith cues to interpret angular motion.