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Vestibular efferent neurons project to the flocculus.

A bilateral projection from the vestibular efferent neurons, located dorsal to the genu of the facial nerve, to the cerebellar flocculus and ventral paraflocculus was demonstrated. Efferent neurons were double-labeled by the unilateral injections of separate retrograde tracers into the labyrinth and into the floccular and ventral parafloccular lobules. Efferent neurons were found with double retrograde tracer labeling both ipsilateral and contralateral to the sites of injection. No double labeling was found when using a fluorescent tracer with non-fluorescent tracers such as horseradish peroxidase (HRP) or biotinylated dextran amine (BDA), but large percentages of efferent neurons were found to be double labeled when using two fluorescent substances including: fluorogold, microruby dextran amine, or rhodamine labeled latex beads. These data suggest a potential role for vestibular efferent neurons in modulating the dynamics of the vestibulo-ocular reflex (VOR) during normal and adaptive conditions.

Animals↗

Visual sensory substitution in vestibular compensation: neuronal substrates in the alert cat.

The purpose of this study was to investigate adaptive changes in the activity of vestibular nuclei neurons unilaterally deprived of their primary afferent inputs when influenced by visual motion cues. These neuronal changes might account for the established role that vision plays in the compensation for posturo-kinetic deficits after the loss of vestibular inputs. Neuronal recordings were made in alert, non-paralysed cats that had undergone unilateral vestibular nerve sections. The unit responses collected in both Deiters' nuclei were compared to those previously recorded in intact cats. We analysed the extracellular activity of Deiters' nucleus neurons, as well as the optokinetic reflex (OKR) evoked during sinusoidal translation of a whole-field optokinetic stimulus in the vertical plane. In intact cats, we found the unit firing rate closely correlated with the visual surround translation velocity, and the relationship between the discharge rate and the motion frequency was tuned around an optimal frequency. The maximum firing rate modulation was generally below the 0.25 Hz stimulus frequency; unit responses were weak or even absent above 0.25 Hz. From the 4th day to the end of the 3rd week after ipsilateral deafferentation, a majority of cells was found to display maximum discharge modulation during vertical visual stimulation at 0.50 Hz, and even at 0.75 Hz, indicating that the frequency bandwidth of the visually induced responses of deafferented vestibular nuclei neurons had been extended. Consequently, the frequency-dependent attenuation in the sensitivity of vestibular neurons to visual inputs was much less pronounced. After the first 3 weeks post-lesion, the unit response characteristics were very similar to those observed prior to the deafferentation. On the nucleus contralateral to the neurectomy, the maximum modulation of most cells was tuned to the low frequencies of optokinetic stimulation, as also seen prior to the lesion. We found, however, a subgroup of cells displaying well-developed responses above 0.50 Hz. Under all experimental conditions, the neuronal response phase still remained closely correlated with the motion velocity of the vertical sinusoidal visual pattern. We hypothesize that Deiters' neurons deprived of their primary afferents may transiently acquire the ability to code fast head movements on the basis of visual messages, thus compensating, at least partially, for the loss of dynamic vestibular inputs during the early stages of the recovery process.(ABSTRACT TRUNCATED AT 400 WORDS)

Adaptation, Physiological↗

Superior vestibular nucleus neurones related to the excitatory vestibulo-ocular reflex of anterior canal origin and their ascending course in the cat.

Stimulation of the superior vestibular nucleus and the anterior canal nerve evoked mono- and disynaptic excitatory postsynaptic potentials, respectively, in contralateral inferior oblique motoneurones of the cat. Combined stimulation revealed that the superior vestibular nucleus relayed excitatory anterior canal signals to the motoneurones. Thirty-six superior vestibular neurones receiving anterior canal inputs were activated antidromically by microstimulation of the contralateral inferior oblique motoneurone pool. Their axons ascended neither in the brachium conjunctivum nor in the medial longitudinal fasciculus, but proceeded rostrally in the ventral part of the brain stem.

Animals↗

Identification of vestibular efferent neurons in the gerbil: histochemical and retrograde labelling.

The efferent neurons of the gerbil vestibular system were investigated by retrograde tracing techniques and cytochemical staining for acetylcholinesterase (AChE), choline acetyltransferase (ChAT) and a number of peptides. The location, bilateral distribution, cell area and number of neurons in two identified groups of retrogradely labelled cells were described and quantified. The larger of the two groups was located dorsolateral to the facial nerve genu, ventral and medial to the vestibular nuclei. Unilateral tracer injection in the vestibular end organs labelled cells bilaterally in this and the smaller group, which was located immediately ventral to the genu. No cells were found that individually projected bilaterally to both labyrinths. After injections of horseradish peroxidase (HRP) in the utricle or saccule, significantly more cells were located on the contralateral side of the brainstem. The average (+/- SD) cross sectional area of labelled cell bodies associated with the otolith organs was 259.8 (+/- 75.2) microns 2. ChAT immunoreactive and AChE positive cells were found in an area coextensive with the location of the dorsal efferent group. In double-labelling studies, cell bodies in the same group that had been retrogradely labelled with a utricular injection of HRP, were immunocytochemically stained for calcitonin gene-related peptide and met-enkephalin. In contrast, the ventral group of efferents did not have cells that were cytochemically stained for either of the acetylcholine-related enzymes or either peptide. The significance of the existence of peptidergic vestibular efferent neurons is discussed.

Acetylcholinesterase↗

Survival of the vestibular nerve after labyrinthectomy in the cat.

Temporal bone studies in cat, monkey, and man demonstrate that the cell bodies of the primary vestibular neurons located in Scarpa's ganglion persist after labyrinthectomy. However, it is not known whether the centrally directed axon process of deafferented vestibular neurons survive or degenerate after labyrinthectomy. If the central axon were to persist, then the primary vestibular neuron could influence vestibular compensation or produce symptoms of vestibular dysfunction. In the present study the temporal bones and brain stem of four cats were prepared for light microscopic examination with hematoxylin-eosin, silver, and trichrome connective tissue stains. Cell counts within Scarpa's ganglion were performed. After labyrinthectomy, many intact axons were demonstrated in the brain stem, a finding that correlated with survival of neurons in Scarpa's ganglion. This study provides anatomic evidence that primary vestibular neurons that survive labyrinthectomy may retain their central axon processes. The persistence of this neural pathway and data from behavioral studies in the cat suggest that vestibular neurons may affect vestibular compensation after labyrinthectomy. Deafferented vestibular neurons may play a role in human vestibular compensation and dysfunction.

Animals↗

Developing vestibular ganglion neurons switch trophic sensitivity from BDNF to GDNF after target innervation.

Recent evidence showing a distinctive cell loss in vestibular and cochlear ganglia of brain-derived neurotrophic factor (BDNF) versus neurotrophin-3 (NT-3) null mutant mice demonstrates that these neurotrophins play a critical role in inner ear development. In this study, biological functions of BDNF and NT-3 in the chick vestibular and cochlear ganglion development was assessed in vitro and compared to those of other neurotrophic factors. The embryonic day (E)8-12 vestibular ganglion neurons showed an extensive outgrowth in response to BDNF with less outgrowth to NT-3. In contrast, NT-3 had stronger neurotrophic effects on the E12 cochlear ganglion neurons compared to BDNF. These results support previous evidence that neurotrophins play important roles in the vestibular and cochlear ganglion neuron development. However, the responsiveness to the neurotrophins declined and became undetectable by E16. Unexpectedly, glial cell line-derived neurotrophic factor (GDNF) promoted neurite outgrowth from vestibular ganglia at E12-16, later than the stages at which BDNF had neurotrophic effects. The time of switching sensitivity of the vestibular ganglion neurons from BDNF to GDNF correlated with the time of completion of synaptogenesis on their peripheral and central targets. Furthermore, a factor released from E12 inner ears exerted neurotrophic effects on late-stage vestibular ganglion neurons that were not responsive to the E4 otocyst-derived factor. These results raise the possibility that the vestibular ganglion neurons become responsive to GDNF upon target innervation and that the changes in sensitivity are regulated by changes in available factors released from their peripheral targets, the inner ear epithelia.

Animals↗

Selective retrograde labelling of vestibular efferent neurons with [3H]choline.

Following administration of [3H]choline in the lateral semicircular canal of the cat labyrinth, bidirectional axoplasmic transport [3H]choline and its derivatives was shown by radioautography in the vestibular system. Light-microscopic radioautographs exhibited various patterns of radioautographic labelling. First, a diffuse reaction was observed in vestibular nuclei representing anterograde-labelled, vestibular nerve endings. Second, a heavy labelling limited to perikarya was detected in efferent vestibular neurons and corresponded to retrograde transport. The anterograde migration of [3H]choline is known to be non-selective and is related to synthesis of phospholipids, non-diffusable molecules. In contrast, the retrograde perikaryal labelling seems highly selective and related to the cholinergic specificity of the transmitter. The selectivity of such labelling offers a further possibility of identifying cholinergic neurons and is additional evidence that cholinergic mechanisms are involved in the efferent vestibular control.

Animals↗

Emergence of action potential generation and synaptic transmission in vestibular nucleus neurons.

Principal cells of the chick tangential nucleus are vestibular nucleus neurons in the hindbrain. Although detailed information is available on the morphogenesis of principal cells and synaptogenesis of primary vestibular fibers, this is the first study of their early functional development, when vestibular terminals emerge at embryonic days 10 and 13 (E10 and E13). At E10, 60% of principal cells generated spikes on depolarization, whereas 50% exhibited excitatory postsynaptic currents (EPSCs) on vestibular-nerve stimulation. The frequency was 0.2 Hz for glutamatergic spontaneous EPSCs (sEPSCs) at -60 mV, and 0.6 Hz for spontaneous inhibitory postsynaptic current (sIPSC) at +10 mV and completely GABAergic. All of these synaptic events were TTX-insensitive, miniature events. At E13, 50% of principal cells generated spikes on depolarization and 82% exhibited EPSCs on vestibular-nerve stimulation. The frequency was 0.7 Hz for sEPSCs at -60 mV, and 0.8 Hz for sIPSCs at +10 mV. Most principal cells had sIPSCs composed of both GABAergic (75%) and glycinergic (25%) events, but a few cells had only GABAergic sIPSCs. TTX decreased the frequency of EPSCs by 12%, and the IPSCs by 17%. In summary, at E10, some principal cells generated immature spikes on depolarization and EPSCs on vestibular-nerve stimulation. At E10, GABAergic events predominated, AMPA events had low frequencies, and glycinergic activity was absent. By E13, glycinergic events first appeared. This data were compared systematically to that obtained from the late-term embryo and hatchling to reveal the long-term sequence of changes in synaptic events and excitability and offer a broader understanding of how the vestibular system is assembled during development.

Action Potentials↗

Role of cross-striolar and commissural inhibition in the vestibulocollic reflex.

In the otolith system, there are two types of neuronal circuitry that can enhance response sensitivity during linear acceleration and tilt of the head. One produces cross-striolar inhibition and the other commissural inhibition. Cross-striolar inhibition can be observed in over 50% of saccular-activated, second-order vestibular neurons. In contrast, it is seen in less than 33% of utricular-activated, second-order vestibular neurons. The majority of vestibular neurons that receive cross-striolar inhibition have axons that project to the spinal cord. Over 50% of the utricular-activated, second-order vestibular neurons received commissural inhibition from the contralateral utricular nerve. On the other hand, almost all the saccular-activated, second-order vestibular neurons exhibit no response to stimulation of the contralateral saccular nerve. The majority of vestibular neurons receiving commissural inhibition also have axons that project to the spinal cord. These results suggest that cross-striolar inhibition is important for increasing the sensitivity of the saccular system, whereas commissural inhibition is more important in the utricular system.

Animals↗

Prednisolone excitation of medial vestibular nucleus neurons in cats.

An electrophysiological study was performed to determine whether prednisolone hydrochloride directly influenced neuronal activities of the medial vestibular nucleus (MVN) in alpha-chloralose-anesthetized cats. Single neuronal activities of MVN were recorded extracellularly with a glass-insulated silver wire microelectrode attached along a seven-barreled micropipette. Each barrel was filled with prednisolone, glutamate, glutamic acid diethylester (GDEE) or CoCl2. Except for prednisolone, which was administered both intravenously and microiontophoretically, other chemicals were applied microiontophoretically to the immediate vicinity of the target neurons. These MVN neurons were classified as type I and II neurons according to their responses to horizontal and sinusoidal rotations. Intravenous prednisolone (up to 5 mg/kg) enhanced spontaneous and rotation-induced neuronal firings of both type I and II neurons in a dose-dependent manner. In a similar tendency, microiontophoretically applied prednisolone (50-200 nA) dose-dependently increased spontaneous and rotation-induced firings of both type I and II neurons. Microiontophoretic GDEE, a non-selective glutamate receptor antagonist, inhibited glutamate- and rotation-induced neuronal discharges without affecting prednisolone-induced increases in neuronal responses of MVN. In addition, iontophoretically applied CoCl2, a Ca2+ channel blocker, did not affect prednisolone-, glutamate- and rotation-induced neuronal findings of MVN. These results suggest that prednisolone induces excitation of type I and II neurons, probably by acting directly on the membrane of MVN neurons. Thus, glucocorticoids such as prednisolone may be effective for the treatment of vertigo resulting from hypofunction of vestibular nucleus neurons.

Animals↗

[Functional characteristics of the input-output correlation in the frog vestibular nuclei].

Field and intracellular potentials were recorded in the vestibular nuclear complex of the frog perfused brain following stimulation of the anterior branch of the ipsilateral vestibular nerve and spinal cord. Mono- and polysynaptic EPSPs with orthodromic APs were recorded from vestibular neurones following vestibular nerve stimulation. Antidromic activation of neurones sending their axons to the labyrinth was also recorded. Antidromic APs of vestibulo-spinal neurones evoked with mean latency of 1.43 and 2.19 ms to stimulation of cervical and lumbar cords, respectively, were revealed.

Action Potentials↗

Ultrastructure of contralaterally projecting vestibular efferent neurons in the cat.

Numerous reports have demonstrated that primary vestibular afferent axons terminate in the vicinity of centrally located vestibular efferent (VE) neurons. The goal of this study was to morphologically demonstrate contact of primary vestibular afferents directly on the soma of contralaterally projecting VE neurons, as well as develop a quantitative description of these neurons. A right vestibular neurectomy was performed in three young adult cats followed up three days later by the placement of horseradish peroxidase into the left vestibule. Labeled contralateral (right) VE neurons were examined in a transmission electron microscope for degenerating synaptic profiles contacting the soma. Ultrastructural characteristics of VE neurons, including nuclear size, volume fraction of intracellular organelles, number of synaptic profiles, and cellular shape, were quantitatively analyzed. No degenerating synaptic profiles were observed on labeled VE soma, although degenerating axons were present nearby. Statistical analysis of volume fraction data of the VE neurons suggests a continuum of a single subpopulation of cells rather than multiple subpopulations.

Animals↗

[Study of vestibular efferent neurons of the guinea pig by the technic of retrograde axonal transport of horseradish peroxidase and with fluorochromes].

Vestibular efferent neurons were identified in different structures of guinea pig medulla using retrograde labelling by horseradish peroxidase and transport-specific fluorochromes. Two groups of vestibular efferent neurons with bilateral projections were found. These groups were situated dorsally and ventrally to the facial nerve fibre pathway. Dorsal group of vestibular efferent neurons consisted of small cells located in close proximity to the genu and the root of facial nerve and in subependymal granular layer of the 4th ventricle. Ventral group comprised middle-size cells usually with weak tracer accumulation; these cells were scattered in vast area of lateral tegmental field within nucleus reticularis lateralis parvocellularis. According to the cell location, morphology and pattern of tracer accumulation a possibility of presence of true vestibular efferents and satellite cells in studied groups is discussed.

Animals↗

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.

Animals↗

Acoustic and vestibular efferent neurons in the chicken (Gallus domesticus). A horseradish peroxidase study.

The origin of labyrinthine efferent fibres was investigated in the chicken. After injection of horseradish peroxidase (HRP) into the cochlear duct or into the ampulla of the horizontal semicircular canal, efferent neurons in the medulla oblongata were labelled by the granular reaction product of retrogradely transported HRP. Efferent vestibular neurons were found in the nucleus reticularis pontis caudalis bilaterally with more cells ipsilateral to the injection side, while efferent cochlear neurons could be demonstrated medial to the ventral division of the facial nucleus bilaterally with more neurons contralateral to the injection.

Animals↗

Morphophysiological study on the divergent projection of axon collaterals of medial vestibular nucleus neurons in the cat.

(1) Spikes of single neurons were extracellularly recorded in the medial vestibular nucleus (MVN) in decerebrate cats and were functionally identified as secondary type I neurons by observing their responses to horizontal rotation and monosynaptic activation after stimulation of the ipsilateral vestibular nerve. Axonal projection of these neurons was examined by their antidromic responses to stimulation of the contralateral abducens nucleus, the spinal cord, and the ascending and descending MLF. (2) Almost all secondary type I vestibular neurons which sent their axon to the contralateral abducens nucleus were antidromically activated from the descending MLF at the level of the obex as well. Nearly half of these neurons sent their collateral axon to the level of C1 segment in the spinal cord and approximately one third to the ascending MLF close to the oculomotor complex. (3) The mean conduction velocity was 29 m/s for descending collateral axons and 30 m/s for ascending collateral axons. (4) Systematic tracking for antidromic microstimulation in the contralateral abducens nucleus and spinal gray matter at C2-C3 suggested that collateral axons of single type I vestibular neurons gave off local branches in the abducens nucleus and the motoneuron pool in the upper cervical gray matter. Existence of terminal branches in the neck motoneuron pool was confirmed by intraaxonal staining with horseradish peroxidase (HRP). (5) Neurons which projected to both the contralateral abducens nucleus and the spinal cord were located in a fairly localized region in the ventrolateral part of the rostral MVN. Neurons which projected to the contralateral abducens nucleus and not to the spinal cord were located in a rostrocaudally wider area in the ventrolateral MVN. Neurons projecting to the spinal cord and not to the contralateral abducens nucleus were located in the widest area in the rostrocaudal direction, covering almost the whole extent of the rostral half of the MVN.

Animals↗

Dopaminergic agonists have both presynaptic and postsynaptic effects on the guinea-pig's medial vestibular nucleus neurons.

A number of studies have indicated a possible interaction between dopamine and the vestibular system. Using intracellular recordings in brainstem slices, we have tested the effects of dopamine and other dopaminergic compounds on guinea-pig medial vestibular nucleus (MVN) neurons. In normal medium, MVN neurons were depolarized by dopamine as well as by (-)quinpirole and piribedil, which are selective D2 dopaminergic agonists. The dependence of this effect on the presence of D2 receptors was confirmed by using (-)sulpiride, a D2 antagonist which blocked the depolarizing effect of dopamine. Dopaminergic D1 receptors were apparently not involved in this effect since a selective D1 agonist, SKF-38393, had no effect on MVN neurons and the D1 antagonist (+)SCH-23390 could not block the effect of dopamine. These depolarizing responses to dopamine must be due to a presynaptic action on terminals that normally release GABA spontaneously on MVN neurons, and tonically maintain them in a state of hyperpolarization. Indeed, such a spontaneous release was demonstrated to occur in the slice since application of bicuculline, a GABAA antagonist, depolarized MVN neurons in normal saline, but not in a high Mg2+/low Ca2+ solution known to block synaptic transmission. When dopamine was applied in conditions in which no GABAA-dependent transmission could occur (either in the presence of bicuculline or in a high Mg2+/low Ca2+ solution) only a hyperpolarizing, most probably postsynaptic, effect occurred. These results indicate that dopamine might exert in vivo a significant modulatory action on the vestibular system, either by a direct action on the vestibular neurons or by modulation of GABAergic transmission.

Animals↗

Basic organization principles of the VOR: lessons from frogs.

Locomotion is associated with a number of optical consequences that degrade visual information processing in the absence of appropriate compensatory movements. The resulting retinal image flow is counteracted by coordinated eye-head reflexes that are initiated by optokinetic and vestibular inputs. The contribution of the vestibulo-ocular reflex (VOR) for stabilizing retinal images is relatively small in amplitude in frogs but important in function by compensating for the non-linearities of the neck motor system. The spatial tuning of the VOR networks underlying the angular (AVOR) and linear (LVOR) with respect to canal and extraocular motor coordinates is organized in a common, canal-related reference frame. Thereby, the axes of head and eye rotation are aligned, principle and auxiliary VOR connections transform vestibular into motor signals and parallel AVOR and LVOR circuits mediate vergence and version signals separately. Comparison of these results with data from other vertebrates demonstrates a number of fundamental organization principles common to most vertebrates. However, the fewer degrees of behavioral freedom of frogs are reflected by the absence of, e.g. a functioning velocity storage network or of a fixation suppression of the VOR. In vitro experiments with the isolated brainstem and branches of N.VIII attached were used to study the putative transmitters of vestibular nerve afferent inputs, the postsynaptic receptor subtypes of second-order vestibular neurons and their dynamic response properties. Evidence is presented that suggests that afferent vestibular nerve fibers with different dynamic response properties activate different subtypes of glutamate receptors. The convergence pattern of monosynaptic afferent nerve inputs from different labyrinthine organs onto second-order vestibular neurons is remarkably specific. As a rule, second-order vestibular neurons receive converging afferent nerve inputs from one semicircular canal and from a specific sector of hair cells on one otolith organ. This convergence pattern remains malleable even in adulthood and reorganization is initiated by activity-related changes in vestibular nerve afferent fibers. The output of second-order vestibular neurons is modified by at least three inhibitory control loops. Uncrossed inhibitory vestibular side loops appear to control specifically the dynamic response tuning, whereas coplanar commissural inhibitory inputs improve mainly the spatial tuning and the cerebellar feedback loop controls the response gain. Among the targets of second-order vestibular projection neurons are extraocular motoneurons and internuclear neurons. Extraocular motoneurons differ among each other by the presence of very different response dynamics. These differences may represent a co-adaptation to the response dynamics of twitch and non-twitch extraocular muscle fibers. Different dynamical properties are required for a rapid acceleration of the globe at the one end and for the maintenance of a stable eccentric eye position over long periods of time at the other end of a continuum of variations in dynamic response properties. The maintenance of a given eccentric eye position over long periods of time is especially well developed in frogs and assists visual surveillance during lurking in the absence of saccades.

Animals↗