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Characterization of spontaneous synaptic transmission in rat medial vestibular nucleus.

Spontaneous synaptic currents of medial vestibular nucleus (MVN) neurons were studied using whole cell recording in slices prepared from 13- to 17-day-old rats. The spontaneous inhibitory postsynaptic currents (sIPSCs) were significantly reduced by the GABAA antagonist bicuculline (20 microM), but were not affected by the glycine antagonist strychnine (1 microM). The frequency, amplitude, and decay time constant of sIPSCs were 4.3 +/- 0.9 Hz, 18.1 +/- 2.0 pA and 8.9 +/- 0.4 ms, respectively. The specific AMPA receptor antagonist GYKI-52466 (50 microM) completely blocked the non-NMDA-mediated spontaneous excitatory postsynaptic currents (sEPSCs), indicating that they are mediated by an AMPA-preferring receptor. The AMPA-mediated sEPSCs were characterized by low frequency (1.5 +/- 0.4 Hz), small amplitude (13.9 +/- 1.9 pA) and rapid decay kinetics (2.8 +/- 0.2 ms). The majority (15/21) displayed linear I-V relationships, suggesting the presence of GluR2-containing AMPA receptors.

2-Amino-5-phosphonovalerate↗

Visual-vestibular interaction in the flocculus of the alert monkey. I. Input activity.

Neuronal activity in the flocculus of alert Rhesus monkeys was recorded during vestibular stimulation (rotation of the monkey about a vertical axis in complete darkness), optokinetic stimulation (rotation of the visual surround around the stationary monkey), combined visual-vestibular stimulation (rotation of the monkey inside the stationary surround in light), and conflicting visual-vestibular stimulation (rotation of the monkey together with the visual surround in the same direction). The input to the flocculus was recorded as non-Purkinje cell (non-P-cell) activity. Ninety per cent of the non-P-cells which were modulated during our stimulation paradigms carry information similar to that in the neurons of vestibular nuclei. This suggests that the main mossy fiber input to the flocculus originates in the vestibular nuclei. A second input of unknown origin conveys visual information about retinal slip. Thus, part of the flocculus -- as further discussed elsewhere (Waespe and Henn 1981) -- may be specialized to subserve visual-vestibular interaction to improve the nystagmus response.

Animals↗

A morphometric and immunohistochemical study of the vestibular nuclear complex in bovine spongiform encephalopathy.

A morphometric and immunohistochemical study of the vestibular nuclear complex was performed on five bovine spongiform encephalopathy (BSE) and five control cow brains. Neurons of the lateral and superior vestibular nuclei were counted at 500-microns intervals in 10-microns-thick sections, using an image analysis system comprising a projection microscope and digitising pad linked to a computer. A bimodal distribution of neuron diameters was recognised in the brains of normal cattle. One population of neurons had a mean diameter of 30 microns and the other had a mean diameter of 60 microns. The vestibular nuclei from BSE cattle had an approximately 50% reduction in total numbers of neurons when compared with controls (P < 0.01). Cattle which were clinically diseased longer had the fewest number of neurons preserved. Diminished numbers of neurons were detected throughout the area studied and affected neurons of all diameters. Immunohistochemical staining for synaptophysin, a protein present in synapses throughout the CNS, showed no significant reduction in axon terminals synapsing with vestibular neurons, including vacuolated neurons of BSE brains, when controls and BSE brains were compared. This suggests that de-afferentation of neurons is not the cause of neuronal loss. Prion protein was detected in the neuropil of the vestibular nuclear complex of BSE brains but not control brains. These studies show that previously unsuspected neuronal loss is a significant feature of BSE.

Animals↗

Comparison of ephrin-A ligand and EphA receptor distribution in the developing inner ear.

Members of the recently discovered Eph family appear to play important roles in a variety of developmental processes including tissue segmentation, cell migration and axonal guidance. To begin to understand the functions of the EphA subclass of receptors and their corresponding GPI-linked (ephrin-A) ligands in the inner ear, a developmental immunohistochemical analysis was completed. The results indicated that the ligands ephrin-A1 and ephrin-A2 were localized mainly at cellular boundaries in the inner ear. Ephrin-A1 was detected mainly in the epithelial cells lining the fluid filled ducts of the inner ear, whereas ephrin-A2 was prominently expressed in connective tissue regions. The receptor EphA4 was detected in vestibular hair cells. EphA5 and EphA7 were detected mainly in cochlear and vestibular supporting cells. These results suggest that these Eph molecules play a role in establishing the formation and cellular organization of the complex inner ear labyrinth. Additionally, all of the ligands and receptors evaluated were expressed in vestibular and cochlear neurons at various developmental stages, suggesting they may play a role in establishing or maintaining innervation to the inner ear.

Actins↗

Distinct distributions of five NMDA receptor channel subunit mRNAs in the brainstem.

The distribution of five NMDA receptor channel subunit mRNAs in the mouse brainstem at postnatal day 21 was semiquantitatively examined by in situ hybridization with subunit-specific oligonucleotide probes. The epsilon 1 subunit mRNA was observed in various brainstem nuclei. On the other hand, the epsilon 2 and epsilon 3 subunit mRNAs were restricted to particular nuclei, and the epsilon 4 subunit mRNA was detected in several nuclei at very low levels. The dorsal cochlear nucleus exhibited differential expression of the epsilon subunit mRNAs in distinct neuron types: the epsilon 2 subunit mRNA in the pyramidal cells, the epsilon 3 subunit mRNA in the granule cells, and the epsilon 1 subunit mRNA in other neurons. In the vestibular nuclei, the medial nucleus expressed the epsilon 1, epsilon 3, and epsilon 4 subunit mRNAs, whereas the lateral and superior nuclei expressed the epsilon 1 subunit mRNA. Such region-specific expressions were also discerned in the central gray, the superior and inferior colliculi, the medial accessory oculomotor nucleus, the locus ceruleus, the parabrachial nucleus, nucleus of the solitary tract, the caudal subnucleus of the trigeminal spinal tract nucleus, and the inferior olive. In contrast to the differential distributions of the four epsilon subunit mRNAs, intense signals for the zeta 1 subunit mRNA were distributed throughout the brainstem. These findings suggest that anatomical organization of the epsilon subunits is heterogeneous in various regions of the brainstem, which would give rise to functional diversity of the NMDA receptor channel in these regions.

Animals↗

A critical period for functional vestibular development in zebrafish.

We have determined a critical period for vestibular development in zebrafish by using a bioreactor designed by NASA to simulate microgravity for cells in culture. A critical period is defined as the briefest period of time during development when stimulus deprivation results in long lasting or permanent sensory deficits. Zebrafish eggs were collected within 3 hours of being laid and fertilized. In experiment 1, eggs were placed in the bioreactor at 3, 24, 30, 36, 48, or 72 hours postfertilization (hPF) and maintained in the bioreactor until 96 hPF. In experiment 2, eggs were placed in the bioreactor immediately after they were collected and maintained in the bioreactor until 24, 36, 48, 60, 66, 72, or 96 hPF. Beginning at 96 hPF, all larvae had their vestibulo-ocular reflexes (VOR) evaluated once each day for 5 days. Only larvae that hatched from eggs that were placed in the bioreactor before 30 hPF in experiment 1 or removed from the bioreactor later than 66 hPF in experiment 2 had VOR deficits that persisted for at least 5 days. These data suggest a critical period for vestibular development in the zebrafish that begins before 30 hPF and ends after 66 hPF. To confirm this, zebrafish eggs were placed in the bioreactor at 24 hPF and removed at 72 hPF. VORs were evaluated in these larvae once each day for 5 days beginning at 96 hPF. These larvae had VOR deficits that persisted for at least 5 days. In addition, larvae that had been maintained in the bioreactor from 24 to 66 hPF or from 30 to 72 hPF, had only temporary VOR deficits. In a final experiment, zebrafish eggs were placed in the bioreactor at 3 hPF and removed at 96 hPF but the bioreactor was turned off from 24 hPF to 72 hPF. These larvae had normal VORs when they were removed from the bioreactor at 96 hPF. Taken as a whole, these data support the idea that there is a critical period for functional maturation of the zebrafish vestibular system. The developmental period identified includes the timeframe during which the vestibular primary afferent neurons are born, innervate their central and peripheral targets, and remodel their central projections.

Animals↗

Associative learning in a network model of Hermissenda crassicornis. II. Experiments.

A companion paper in a previous issue of this journal presented a resistance-capacitance circuit computer model of the four-neuron visual-vestibular network of the invertebrate marine mollusk Hermissenda crassicornis. In the present paper, we demonstrate that changes in the model's output in response to simulated associative training is quantitatively similar to behavioral and electrophysiological changes in response to associative training of Hermissenda crassicornis. Specifically, the model demonstrates many characteristics of conditioning: sensitivity to stimulus contingency, stimulus specificity, extinction, and savings. The model's learning features also are shown to be devoid of non-associative components. Thus, this computational model is an excellent tool for examining the information flow and dynamics of biological associative learning and for uncovering insights concerning associative learning, memory, and recall that can be applied to the development of artificial neural networks.

Animals↗

Nystagmus induced by stimulation of the nucleus of the optic tract in the monkey.

1. The nucleus of the optic tract (NOT) was electrically stimulated in alert rhesus monkeys. In darkness stimulation evoked horizontal nystagmus with ipsilateral slow phases, followed by after-nystagmus in the same direction. The rising time course of the slow phase velocity was similar to the slow rise in optokinetic nystagmus (OKN) and to the charge time of optokinetic after-nystagmus (OKAN). The maximum velocity of the steady state nystagmus was approximately the same as that of OKAN, and the falling time course of the after-nystagmus paralleled OKAN. 2. Increases in frequency and duration of stimulation caused the rising and falling time constants of the nystagmus and after-nystagmus to become shorter. Changes in the falling time constant of the after-nystagmus were similar to changes in the time constant of OKAN produced by increases in the velocity or duration of optokinetic stimulation. 3. Stimulus-induced nystagmus was combined with OKN, OKAN and per- and post-rotatory nystagmus. The slow component of OKN as well as OKAN could be prolonged or blocked by stimulation, leaving the rapid component of OKN unaffected. Activity induced by electrical stimulation could also sum with activity arising in the semicircular canals to reduce or abolish post-rotatory nystagmus. 4. Positive stimulus sites for inducing nystagmus were located in the posterolateral pretectum. This included portions of NOT that lie in and around the brachium of the superior colliculus and adjacent regions of the dorsal terminal nucleus (DTN). 5. The data indicate that NOT stimulation had elicited the component of OKN which is responsible for the slow rise in slow phase velocity and for OKAN. The functional implication is that NOT, and possibly DTN, are major sources of visual information related to retinal slip in the animal's yaw plane for semicircular canal-related neurons in the vestibular nuclei. Analyzed in terms of a model of OKN and OKAN (Cohen et al. 1977; Waespe et al. 1983), the indirect pathway, which excites the velocity storage mechanism in the vestibular system to produce the slow component of OKN and OKAN, lies in NOT in the monkey, as it probably also does in cat, rat and rabbit. Pathways carrying activity for the rapid rise in slow phase velocity during OKN or for ocular pursuit appear to lie outside NOT.

Animals↗

Activation of the efferent system in the isolated frog labyrinth: effects on the afferent EPSPs and spike discharge recorded from single fibres of the posterior nerve.

Intra-axonal recordings were obtained from single afferent fibres of the posterior nerve in the isolated labyrinth of the frog (Rana esculenta). EPSPs and spike discharge were recorded both at rest and during rotatory stimulation of the canal. Electrical stimulation of either the distal end of the cut posterior nerve or of the central stumps of the anterior-horizontal nerves elicited a frequency-dependent inhibitory effect on the afferent discharge arising from the posterior canal. Denervation experiments revealed that inhibition is mediated by efferent fibres exhibiting a high degree of branching in the proximal part of the eighth nerve. The inhibitory effect was selectively cancelled by (1)D-tubocurarine 10(-6) M; (2) atropine 5 x 10(-5) M; (3) acetylcholine or carbachol 10(-4) M; (4) eserine 10(-5) M. Inhibition is thus most likely to be sustained by the release of acetylcholine from the efferent nerve terminals. Experiments in which the ionic composition of the external medium was modified suggest that the transmitter acts mainly by opening the chloride ion channels of the hair cell membrane. In some units the same stimulation pattern evoked a consistent increase in both EPSP and spike discharge, instead of inhibition. Such facilitation was unaffected by drugs for ionic modifications which block the efferent synapse, but disappeared after denervation. Inhibition and facilitation, therefore, act as two control mechanisms which are able to modify substantially, at the first stage of processing, the sensory information which is sent to the vestibular second order neurons.

Acetylcholine↗

Projections from brainstem nuclei to the nucleus paragigantocellularis lateralis in the cat.

Afferent projections from the pons and medulla to the nucleus paragigantocellularis lateralis (PGL) have been mapped in the cat using retrograde transport of horseradish peroxidase (HRP). In the caudal medulla, the major sources of afferents were the medial and lateral divisions of the solitary nuclei complex and the contralateral trigeminal nucleus caudalis. Labelled cells were also present in the dorsal column nuclei, nucleus intercalatus and praepositus hypoglossi but this may have been due to uptake of HRP into fibres of passage. In the dorsolateral medulla and pons, neurones in the vestibular complex and in the parabrachial nucleus were labelled bilaterally. Nucleus raphe magnus and raphe obscurus were both found to send projections to the PGL and labelled cells were also present throughout the pontine and medullary reticular nuclei as well as in PGL on the side opposite to the injection of HRP. These findings are discussed in relation to the role of the PGL in cardiovascular regulation and in the control of pain.

Animals↗

Failure of the oculomotor neural integrator from a discrete midline lesion between the abducens nuclei in the monkey.

Recent anatomical studies indicate that axons of neurons in the vestibular nuclei, projecting to the contralateral abducens nuclei, cross the midline at the abducens level. These axons then give off collaterals to the contralateral vestibular and prepositus nuclei that may be important for the neural integrator that converts eye-velocity to eye-position signals. We disrupted a subset of these commissural projections by making a small midline lesion between the abducens nuclei in a monkey. The vestibulo-ocular reflex and saccades were still present post-lesion, indicating that premotor drive was intact, but the lesion produced severe post-saccadic drift, indicating failure of the neural integrator. We conclude that commissural projections crossing at the abducens level may be important for oculomotor integration.

Abducens Nerve↗

Cytochrome oxidase histochemistry in Scarpa's ganglion after hemilabyrinthectomy.

Cytochrome oxidase histochemistry was studied in neurons in the vestibular ganglion in gerbils two weeks after hemilabyrinthectomy. This study measured the staining density in ganglion cells on both the lesioned and non-lesioned side of the brainstem. Cytochrome oxidase staining was significantly reduced in ganglion cells ipsilateral to the lesion. This decrease may have been related to the concomitant loss of spontaneous discharge and reduced energy demand for oxidative metabolism.

Animals↗

Modulatory roles of GABAergic mechanisms in post-rotatory nystagmus in the rabbit.

1. To determine how the GABAergic mechanism operates in the generation of post-rotatory nystagmus, an experiment was performed with GABAergic drugs in rabbits. 2. Subconvulsive doses of picrotoxin (0.3-0.6 mg/kg, i.v.) and bicuculline (0.1 mg/kg, i.v.) decreased the number of post-rotatory nystagmus beats, whereas strychnine sulphate, at a subconvulsive dose (0.1 mg/kg i.v.), increased it. 3. Diazepam (1 mg/kg, i.v.) remarkably increased the number of post-rotatory nystagmus beats. Pretreatment with picrotoxin (0.45 mg/kg, i.v.), bicuculline (0.1 mg/kg, i.v.) or semicarbazide-HCl (180 mg/kg, i.v.) antagonized the effects of diazepam (1 mg/kg, i.v.). 4. GABAergic mechanisms may play a modulatory role in the production of nystagmus rhythm. Strychnine-sensitive neurons involved in the vestibular mechanism may behave in a different manner from picrotoxin-sensitive neurons.

Animals↗

The role of neurotrophic factors in regulating the development of inner ear innervation.

Several neurotrophins and their receptors regulate the survival of vestibular and cochlear neurons and probably also the efferent and autonomic neurons that innervate the inner ear. Mice lacking either brain-derived neurotrophic factor (BDNF) or its associated receptor, TrkB, lose all innervation to the semicircular canals and have reduced innervation of the outer hair cells in the apical and middle turns of the cochlea. Mice lacking neurotrophin-3 (NT-3) or its receptor, TrkC, lose many spiral ganglion cells predominantly in the basal turn of the cochlea. Nerve fibers from spiral ganglion cells in the middle turn extended to inner hair cells of the base. In mice lacking both BDNF and NT-3, or both TrkB and TrkC, there is a complete loss of innervation to the inner ear. Thus, these two neurotrophins and their associated receptors have been shown to be absolutely necessary for the normal development of afferent innervation of the inner ear. Current research efforts are testing the therapeutic potential for neurotrophins to treat hearing loss.

Animals↗

Characterization of the deafwaddler mutant of the rat plasma membrane calcium-ATPase 2.

The deafwaddler mutant in mice was the first spontaneous mutant discovered in the plasma membrane Ca(2+) pump (PMCA) [Street, V.A. et al., 1998, Nat. Genet. 19, 390-394]. A nucleotide substitution in deafwaddler results in a Gly to Ser transition at amino acid 283 in the small cytoplasmic loop of PMCA isoform 2 (PMCA2). PMCA2 is abundant in the stereocilia of auditory and vestibular hair cells, neurons of the spiral ganglion, and participates in inner ear development. Mice that are homozygous for deafwaddler are deaf and have poor balance. However, the balance and hearing disorders of the deafwaddler mice appear to be less severe than homozygotes for a functionally null frameshift mutant or homozygous PMCA2 knockout mice, suggesting that deafwaddler PMCA2 retains some biological activity. To examine the enzymic effects of the deafwaddler mutant, PMCA2 wild-type and deafwaddler were produced by transient expression in COS cells as well as baculovirus-mediated expression in Sf9 insect cells. Membrane preparations were assayed for calcium transport and ATPase activity. No significant differences in the regulation by calmodulin of the wild-type and deafwaddler PMCA2b were found. Steady-state transport assays and pre-steady-state ATPase assays of these two proteins revealed that the K(0.5) for Ca(2+), K(0.5) for calmodulin, degree of activation by calmodulin and rate of activation by Ca-calmodulin were nearly identical. However, calcium transport of the deafwaddler pump was reduced to 30% of the wild-type activity. Although calcium transport activity was reduced in the deafwaddler pump, total phosphoenzyme formation from ATP was slightly higher for deafwaddler than for wild-type. 50 microM LaCl3 (which blocks the E(1)P to E(2)P conformational transition) increased the steady-state level of phosphoenzyme 3-fold for the wild-type but had no effect on the deafwaddler. Taken together, the kinetic data suggest that the deafwaddler mutation affects PMCA2 by slowing the E(1)P to E(2)P transition, resulting in approximately 70% reduction in the PMCA2-mediated Ca(2+) export.

Adenosine Triphosphate↗

Interaural translational VOR: suppression, enhancement, and cognitive control.

We investigated the influence of cognitive factors on the early response of the interaural translational vestibuloocular reflex (tVOR) in six normal subjects. Variables were prior knowledge of direction of head motion and the position of the fixation target relative to the head [head-fixed (HF) or space-fixed (SF)]. A manually driven device provided a step-like head translation (approximately 35 mm distance, peak acceleration, 0.6-1.3 g). Subjects looked at the SF or HF target located 15 cm in front of their heads in otherwise complete darkness. The testing paradigms were: random interleaving of SF and HF targets with unknown direction of head movement, known target location with random head direction (SFR or HFR), and known target location with known head direction (SFP or HFP). Timing was always unpredictable. A "gain" of the slow phase was calculated with respect to ideal performance (maintained fixation of the SF target, recorded/ideal eye velocity computed at time of peak head velocity). At such times, there were no significant differences in gain between HF and SF trials in the random condition; the average gain was approximately 36% of ideal. On the other hand, responses in the SFR and HFR conditions differed as early as 20 ms after the head began moving. Average gain was higher (0.43 +/- 0.11 vs. 0.34 +/- 0.14; means +/- SD, P < 0.05) for each subject in the SFR than the HFR condition. For SFP and HFP, the responses differed from the onset of head motion. Average slow-phase gain was higher (0.49 +/- 0.12 vs. 0.31 +/- 0.12, P < 0.02) for each subject in SFP than in HFP. The timing of corrective saccades during the tVOR was also influenced by cognitive factors. Visual error signals seemed to be more important for triggering saccades in HF trials, whereas preprogramming, probably based on labyrinthine information, seemed to be more important in SF trials. Simulations showed that the changes in slow-phase gain with cognition could be reproduced with simple parametric adjustments of the gain of activity from otolith afferents and suggest that higher-level cognitive control of the VOR could occur as early as the synapse of peripheral afferents on neurons in the vestibular nuclei, either directly from higher level centers or via the cerebellum. In sum, the tVOR-both in its slow-phase response and the saccadic corrections-is subject to "higher-level" cognitive influences including knowledge of where the line of sight must point during head motion and the impending direction of head motion.

Acceleration↗

Vestibuloocular reflex signal modulation during voluntary and passive head movements.

The vestibuloocular reflex (VOR) effectively stabilizes the visual world on the retina over the wide range of head movements generated during daily activities by producing an eye movement of equal and opposite amplitude to the motion of the head. Although an intact VOR is essential for stabilizing gaze during walking and running, it can be counterproductive during certain voluntary behaviors. For example, primates use rapid coordinated movements of the eyes and head (gaze shifts) to redirect the visual axis from one target of interest to another. During these self-generated head movements, a fully functional VOR would generate an eye-movement command in the direction opposite to that of the intended shift in gaze. Here, we have investigated how the VOR pathways process vestibular information across a wide range of behaviors in which head movements were either externally applied and/or self-generated and in which the gaze goal was systematically varied (i.e., stabilize vs. redirect). VOR interneurons [i.e., type I position-vestibular-pause (PVP) neurons] were characterized during head-restrained passive whole-body rotation, passive head-on-body rotation, active eye-head gaze shifts, active eye-head gaze pursuit, self-generated whole-body motion, and active head-on-body motion made while the monkey was passively rotated. We found that regardless of the stimulation condition, type I PVP neuron responses to head motion were comparable whenever the monkey stabilized its gaze. In contrast, whenever the monkey redirected its gaze, type I PVP neurons were significantly less responsive to head velocity. We also performed a comparable analysis of type II PVP neurons, which are likely to contribute indirectly to the VOR, and found that they generally behaved in a quantitatively similar manner. Thus our findings support the hypothesis that the activity of the VOR pathways is reduced "on-line" whenever the current behavioral goal is to redirect gaze. By characterizing neuronal responses during a variety of experimental conditions, we were also able to determine which inputs contribute to the differential processing of head-velocity information by PVP neurons. We show that neither neck proprioceptive inputs, an efference copy of neck motor commands nor the monkey's knowledge of its self-motion influence the activity of PVP neurons per se. Rather we propose that efference copies of oculomotor/gaze commands are responsible for the behaviorally dependent modulation of PVP neurons (and by extension for modulation of the status of the VOR) during gaze redirection.

Animals↗

Asymmetry of the pitch vestibulo-ocular reflex in patients with cerebellar disease.

Responses to pitch head impulses were measured in 15 patients with cerebellar degeneration and downbeat nystagmus and in 5 control subjects, using three-axis search coils. For each subject, response gains were calculated as (1) the ratio of instantaneous vertical eye velocity to pitch head velocity (at 30 ms and at 70 ms into the response), and (2) the ratio of peak eye velocity to peak head velocity. Gains varied more widely among patients. When calculated at 70 ms and using peak values, patients had higher gains for downward pitch, and normal subjects had symmetric gains. At 30 ms, gains were more symmetric in patients, but at that point control subjects actually had lower gains for downward pitch. Thus, the pitch gain ratio (ratio of downward gain to upward gain) was consistently greater in patients than in normal subjects. Because downward impulses were generally faster, eye velocities during downward and upward pitch for equivalent head speeds were also compared. Again, patients had higher gains for downward pitch. These results are consistent with the authors' hypothesis that cerebellar disease results in a higher sensitivity of anterior than posterior semicircular canal pathways, perhaps through loss of inhibition from the flocculus/paraflocculus complex on anterior canal secondary neurons in the vestibular nuclei. The pitch gain asymmetry was larger than, and did not correlate with, the velocity of spontaneous upward drift. This supports the notion that other mechanisms are likely to contribute to downbeat nystagmus in these patients.

Adult↗

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