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Electrophysiological properties of the somatotopic organization of the vestibulospinal system in the frog.

In experiments on the preparation of a frog perfused brain (Rana ridibunda), field and intracellular potentials were recorded from neurons of the vestibular nuclear complex following stimulation of the ipsilateral vestibular nerve and different levels of the spinal cord. Stimulation of the vestibular nerve evoked mono- and polysynaptic excitatory postsynaptic potentials and orthodromic action potentials. In parallel, an antidromic activation of vestibular neurons sending their axons to the labyrinth was recorded. Vestibulospinal neurons sending their axons to the cervical (C neurons) and lumbar (L neurons) enlargements of the spinal cord were identified by their antidromic activation. A rather high conduction velocity along vestibulospinal fibres (mean 15.47 m/s) was observed. A somatotopic arrangement of the vestibulospinal system was established in spite of extremely large overlapping zones for the fore- and hindlimb representations in the vestibular nuclear complex. The hindlimbs were represented more poorly than the forelimbs. Antidromic potentials of C and L neurons were recorded in the medial, descending and with the highest density in the lateral vestibular nuclei (Deiters' nucleus). C neurons were evenly distributed in the other vestibular nuclei studied, while L neurons were located predominantly in the caudal parts of the vestibular nuclear complex. The multiplicity of the origin of the vestibulospinal axons was established. Peculiarities of the functional correlation between the vestibular input and vestibulospinal system are discussed.

Animals↗

Excitation and inhibition of rat medial vestibular nucleus neurones by 5-hydroxytryptamine.

The effects of 5-hydroxytryptamine (5-HT) and related compounds on the discharge rate of tonically active medial vestibular nucleus (MVN) neurones were studied in an in vitro slice preparation of the dorsal brainstem of the rat. The majority (87 of 107, 82%) of MVN neurones were excited by 5-HT. Nine cells (8%) showed a biphasic response to 5-HT, which consisted of a brief inhibition followed by excitation. Eleven cells (10%) were inhibited by 5-HT. The excitatory effects of 5-HT were mimicked by alpha-methyl-5-HT and antagonised by ketanserin and ritanserin, indicating the involvement of the 5-HT2 subtype of 5-HT receptor. In biphasic cells, blockade of 5-HT2 receptors by ketanserin reduced the excitatory component of the response and revealed an enhanced initial inhibition. The inhibitory effects in biphasic cells, and in cells that showed a pure inhibition in response to 5-HT, were blocked by pindobind-5-HT and mimicked by 8-hydroxy-2-(di-n-propylamino)-tetralin indicating the involvement of 5-HT1A receptors. The significance of these findings in relation to the effects of 5-HT on vestibular reflex function is discussed.

8-Hydroxy-2-(di-n-propylamino)tetralin↗

Evidence for inhibitory amino acid receptors on guinea pig medial vestibular nucleus neurons in vitro.

There is little evidence to indicate the identity of the inhibitory receptors which mediate inhibitory interaction between the two medial vestibular nuclei ('brainstem commissural inhibition'). In the present study we tested the hypothesis that medial vestibular nucleus (MVN) neurons have gamma-aminobutyric acid (GABA) or glycine receptors by recording from single MVN neurons in isolated guinea pig MVN slices maintained in vitro while superfusing with GABA (10(-8) M) and the non-competitive GABAA antagonist picrotoxin (10(-6) M or 2 x 10(-6) M), or glycine (10(-6) M) and the competitive glycine antagonist strychnine (10(-6) M). Forty-four % (16/36) of the neurons tested with GABA showed a decrease in firing; in 7 out of 8 cases in which a decrease in firing occurred, the addition of the antagonist picrotoxin completely blocked the effect of the GABA alone. Fifty % (7/14) of the neurons tested with glycine showed a decrease in firing; in 4 out of 6 cases where a decrease occurred, the addition of the antagonist strychnine completely blocked the effect of the glycine alone. In one case only did a cell respond both to GABA and glycine (8 neurons tested with both). These results are consistent with the hypothesis that some MVN neurons have GABA or glycine receptors (but in most cases not both), which may mediate brainstem commissural inhibition.

Animals↗

Histamine depolarizes rat medial vestibular nucleus neurons recorded intracellularly in vitro.

The effects of histamine (HA) on the resting membrane potential and input resistance of rat medial vestibular nucleus (MVN) neurons were investigated using intracellular recording techniques from a submerged brain slice preparation. The exogenous application of HA predominantly produced a concentration-dependent membrane depolarization and induction of action potential firing. The depolarization exhibited a rapid onset, a slow recovery, and usually occurred in the absence of any apparent change in conductance. These effects of HA could be mimicked by the H2-agonist impromidine and were reversibly blocked by the H2-antagonist cimetidine. Tetrodotoxin (TTX) or low calcium/high magnesium-containing media failed to block completely the HA-induced depolarization supporting a direct postsynaptic receptor mediated action of HA. The diminished HA-induced depolarization observed following pretreatment with TTX cannot exclude an additional presynaptic action by HA. The present findings reveal that HA exerts a novel direct excitation of rat MVN neurons through an H2-receptor.

Action Potentials↗

Topological and developmental gradients of calbindin expression in the chick's inner ear.

Mobile intracellular calcium buffers play an important role in regulating calcium flux into mechanosensory hair cells and calbindin D-28k is expressed at high levels in the chick's basilar papilla. We have used RT-PCR, in situ hybridization, and immunohistology to demonstrate that calbindin expression varies systematically according to hair cell position and developmental age. RT-PCR using microdissected quarters of the posthatch basilar papilla showed that mRNA levels were lowest in the (low frequency) apex and higher in basal quadrants. In situ hybridization revealed calbindin mRNA in posthatch hair cells and supporting cells, with more intense labeling of hair cells from basal (high frequency) positions. A similar topology was obtained with calbindin antibodies. Neither calbindin riboprobe nor calbindin antibody labeled cochlear neurons. In contrast, a subset of large vestibular neurons and their calyciform endings onto Type I vestibu lar hair cells were strongly labeled by the calbindin antibody, while vestibular hair cells were negative for calbindin immunoreactivity. Likewise, calbindin in situ hybridization was negative for vestibular hair cells but positive in a subset of larger vestibular neurons. Calbindin mRNA was detected in hair cells of the basal half of the papilla at embryonic day 10 (E10) and calbindin immunoreactivity was detected at E12. Hair cells in the apical half of the papilla had equivalent calbindin expression two days later. Immunoreactivity appeared in abneural supporting cells days later than in hair cells, and not until E20 in neurally located supporting cells. These results demonstrate that calbindin message and protein levels are greater in high-frequency hair cells. This "tonotopic" gradient may result from the stabilization of a basal-to-apical developmental gradient and could be related at least in part to calcium channel expression along this axis.

Aging↗

Coexistence of acetylcholine and calcitonin gene-related peptide in the vestibular efferent neurons in the rat.

An immunocytochemical study combined with the retrograde tracer technique was performed in the rat to assess the vestibular efferent system. All 3 neuron groups which give rise to axons terminating on the vestibular end-organs were cholinergic, i.e., a group dorsolateral to the genu of the facial nerve (DL), a group dorsomedial to this genu (M), and scattered cells in the parvocellular reticular nucleus (PCRt). In addition, we further demonstrated that about 55% of the cholinergic cells in DL had calcitonin gene-related peptide.

Acetylcholine↗

[Analysis of spontaneous unit activity of the lateral vestibular nucleus' neurons in norm and after effects of vibration].

Changes of spontaneous unit activity in the lateral vestibular nucleus of the rat following 5-, 10- and 15-day vibration (60 Hz, 2 hrs. Daily), were studied. Averaged histogramz and autocorrelograms were analysed following computerized interspike intervals. The data obtained revealed a variety of the unit activity in its direct or mediated effects exerted through several structures of the central nervous system.

Animals↗

Opioid inhibition of rat medial vestibular nucleus neurones in vitro and its dependence on age.

Extracellular and whole-cell patch clamp intracellular recordings were made from rat medial vestibular nucleus (MVN) neurones in vitro, and their responses to selective mu-, kappa- and delta-opioid receptor agonists and antagonists were examined. Of 127 neurones tested, the large majority were inhibited in a dose-dependent manner by the delta-opioid receptor agonists [D-Ala2, D-Leu5]-enkephalin (DADLE) and [D-Pen2, Pen5]-enkephalin (DPLPE). The mu-opioid receptor agonist morphine and the kappa-receptor agonist U50,488 did not affect the tonic discharge rate of any of the 63 MVN cells tested. The delta-receptor antagonist naltrindole effectively antagonised the inhibitory effects of DADLE and DPLPE. Weak excitatory responses to high doses of DADLE were seen in only two MVN cells. These results demonstrate the presence of delta- but not mu- or kappa-opioid receptors on tonically active MVN neurones. Whole-cell intracellular recordings from MVN cells in a current clamp showed that the DADLE-induced inhibition was accompanied by membrane hyperpolarisation and decrease in input resistance, while voltage clamp experiments showed that DADLE induced an outward membrane current that was reduced but not abolished by 20 mM tetraethylammonium bromide. Thus the mechanisms of action of DADLE in inhibiting MVN cells involve the potentiation of outward K currents, in a similar way to the effects of opioids in other areas of brain. The inhibitory effects of DADLE increased linearly with age, so that the responses to DADLE in the youngest animals used here (60-80 g, approx. 3 weeks of age) were relatively small, increasing significantly over the following 2-3 weeks. This age-dependence may be due to post-natal changes in the density of delta-opiate receptors or the efficacy of the signalling pathways activated by them in the MVN cells over this time.

Aging↗

Olivocochlear and vestibular efferent neurons of the feline brain stem: their location, morphology and number determined by retrograde axonal transport and acetylcholinesterase histochemistry.

Anterograde degeneration studies have shown that the cochlear and vestibular receptor organs receive an efferent innervation from neurons in the brain stem. This pathway may provide a mechanism by which the CNS could modulate its own afferent input. The neurons which provide this innervation have so far escaped positive identification with methods which depend on retrograde cell changes after axotomy. In the present study, horseradish peroxidase (HRP) was injected into the labryinths of kittens and after allowing 24 hours for the retrograde axonal transport of this tracer, its presence in neurons of the brain stem was demonstrated histochemically. Because there is evidence that the efferent innervation of the labyrinth is cholinergic, acetylcholinesterase (AChE) was also demonstrated histochemically in the same or in adjacent tissue sections. Neurons labelled with HRP were found bilaterally in most periolivary cell groups of the superior olivary complex (cochlear efferents) and in the parvocellular reticular nucleus lateral to the abducens nucleus (vestibular efferents). Counts of labelled neurons yielded estimated totals of 1,700-1,800 cochlear and 400-500 vestibular efferent neurons. Approximately 60% of the neurons in each total were located on the side ipsilateral to the injection. The distribution of HRP-labelled neurons was virtually identical to that of AChE-positive neurons found in adjacent sections, and in those regions with predominantly ipsilateral or contralateral projections, there was an approximate correspondence in number of HRP- and AChE-positive neurons. In tissue sections processed successively for demonstration of HRP and AChE, virtually all HRP-labelled neurons were found to be AChE-positive. These findings suggest that a number of current conceptions regarding labyrinthine efferent systems may need revision.

Acetylcholinesterase↗

Effects of histamine and betahistine on rat medial vestibular nucleus neurones: possible mechanism of action of anti-histaminergic drugs in vertigo and motion sickness.

The tonic discharge of 71 medial vestibular nucleus (MVN) neurones was recorded in slices of the dorsal brainstem of young adult rats. Bath application of histamine caused a dose-related excitation in 59 of the 71 cells (83%), the remaining 12 (17%) being unresponsive. Dimaprit, a selective H2 agonist, also caused excitation in all 20 cells tested. The histamine-induced excitation and the response to dimaprit were antagonised by the selective H2 antagonist ranitidine, confirming that the H2 subtype of histamine receptor is involved in mediating the effects of histamine on these cells. Triprolidine, a selective H1 antagonist, also antagonised the excitation caused by histamine, at a concentration (0.3 microM) which left the H2 receptor-mediated response to dimaprit unchanged. Thus the excitatory effects of histamine on MVN cells in the rat involve two components mediated through H1 and H2 receptor-linked mechanisms, respectively. Betahistine, a weak H1 agonist and H3 antagonist, had little excitatory action when applied on its own, but significantly reduced the excitation caused by histamine when the two drugs were applied together. The effects of betahistine were consistent with a partial-agonist action at H1 receptors on MVN cells, reducing the excitatory responses to histamine presumably by occupying these receptor sites in competition with the exogenously applied neurotransmitter. This partial-agonist action of betahistine may be an important part of its mechanism of action in the symptomatic treatment of vertigo and motion sickness, since it is likely to occur not only in the MVN but also in many brain regions, including the thalamus and cortex, which express H1 receptors and which are innervated by the hypothalamic histaminergic system. Thus the effectiveness of betahistine and other anti-H1 drugs against motion sickness may be explained by their action in reducing the effects of the excess histamine release induced in such conditions in various brain areas, including the MVN.

Animals↗

Vestibular neuritis: an overview using a classical case.

Although acute unilateral and/or bilateral vestibular paralysis, known as vestibular neuronitis, is the second most common cause of vertigo (the most common is benign paroxysmal positional vertigo (BPPV), it is fraught with controversies. The clinical symptoms and methods of treatment of vestibular neuronitis are well defined; however, the etiology and pathophysiology of this disorder are still sketchy. Furthermore, there are no specific diagnostic tests available, and unfortunately, there are no animal models for this disorder. The purpose of this paper is to present an overview of those controversies using a classical case of vestibular neuronitis.

Adult↗

Modulation of rat medial vestibular nucleus neurone activity by vasopressin and noradrenaline in vitro.

In the present study, we examined the effects of bath application of vasopressin and noradrenaline on the spontaneous tonic discharge of medial vestibular nucleus (MVN) neurones and investigated if there is an interaction between the two drugs in an in vitro slice preparation of the rat brainstem containing the MVN. The results showed that vasopressin did not affect the spontaneous discharge rate of MVN neurones when applied either as a 60 s pulse or when the drug continuously perfused the slice for a period of 10 min. In contrast, noradrenaline affected the spontaneous discharge rate of the majority of cells tested (53/60, 88%). Noradrenaline excited the majority (46/53, 87%) of MVN neurones through both alpha1 and beta noradrenergic receptor-linked mechanisms. The remaining cells (7/53, 13%) were inhibited by noradrenaline through an alpha2 noradrenergic receptor-linked mechanism. Neither the excitatory nor inhibitory effects of noradrenaline were modified by vasopressin when the two drugs were applied together.

Action Potentials↗

Vestibular signals in the parasolitary nucleus.

Vestibular primary afferents project to secondary vestibular neurons located in the vestibular complex. Vestibular primary afferents also project to the uvula-nodulus of the cerebellum where they terminate on granule cells. In this report we describe the physiological properties of neurons in a "new" vestibular nucleus, the parasolitary nucleus (Psol). This nucleus consists of 2,300 GABAergic neurons that project onto the ipsilateral inferior olive (beta-nucleus and dorsomedial cell column) as well as the nucleus reticularis gigantocellularis. These olivary neurons are the exclusive source of vestibularly modulated climbing fiber inputs to the cerebellum. We recorded the activity of Psol neurons during natural vestibular stimulation in anesthetized rabbits. The rabbits were placed in a three-axis rate table at the center of a large sphere, permitting vestibular and optokinetic stimulation. We recorded from 74 neurons in the Psol and from 23 neurons in the regions bordering Psol. The activity of 72/74 Psol neurons and 4/23 non-Psol neurons was modulated by vestibular stimulation in either the pitch or roll planes but not the horizontal plane. Psol neurons responded in phase with ipsilateral side-down head position or velocity during sinusoidal stimulation. Approximately 80% of the recorded Psol neurons responded to static roll-tilt. The optimal response planes of evoked vestibular responses were inferred from measurement of null planes. Optimal response planes usually were aligned with the anatomical orientation of one of the two ipsilateral vertical semicircular canals. The frequency dependence of null plane measurements indicated a convergence of vestibular information from otoliths and semicircular canals. None of the recorded neurons evinced optokinetic sensitivity. These results are consistent with the view that Psol neurons provide the vestibular signals to the inferior olive that eventually reached the cerebellum in the form of modulated climbing fiber discharges. These signals provide information about spatial orientation about the longitudinal axis.

Afferent Pathways↗

Electrophysiological and pharmacological characteristics of ionotropic glutamate receptors in medial vestibular nucleus neurons: a whole cell patch clamp study in acutely dissociated neurons.

A patch clamp study was performed to determine which subtype of ionotropic glutamate receptors is involved in the glutamate-induced excitation of the medial vestibular nucleus (MVN) neurons. Whole cell recording was performed on MVN neurons that were acutely dissociated by enzymatic and mechanical treatments. Application of glutamate at a concentration of 100 microM produced a current with a reversal potential of approximately 0 mV. The glutamate-induced current was completely blocked by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM), a non-N-methyl-D-aspartate (NMDA)-receptor antagonist. Application of alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA) and kainic acid (KA), non-NMDA-receptor agonists, at concentrations of 30 and 100 microM produced a concentration-dependent depolarization concomitantly with an increase in firing rates during current clamp recording. During voltage clamp recording, glutamate, AMPA and KA elicited a concentration-dependent current with an equilibrium potential of approximately 0 mV. To clarify whether NMDA receptors are present in MVN neurons, the effects of glycine on the glutamate- and NMDA-induced current were examined. Two types of NMDA receptor-mediated current (types 1 and 2) were obtained in terms of the difference in sensitivity to both magnesium ion and MK-801, which act on the NMDA-receptor channel. In the type 1 neurons, the NMDA-induced current was not apparently blocked by magnesium ion or MK-801, although a larger current was obtained in the absence of magnesium ion. In the type 2 neurons, marked blockade of the NMDA-induced current was seen in the presence of magnesium ion and MK-801, as previously reported in other neurons of the central nervous system. These findings indicate the presence of both non-NMDA and NMDA receptors, which are involved in primary afferent transmission, in the MVN neuron, and two distinct types of NMDA receptors.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

5-HT1A receptor-mediated inhibition of lateral vestibular nucleus neurons projecting to the abducens nucleus.

Electrophysiological studies were performed using cats anesthetized with alpha-chloralose, to elucidate the 5-hydroxytryptamine (5-HT) receptor subtypes involved in the 5-HT-induced inhibition of the lateral vestibular nucleus (LVN) neurons projecting to or through the abducens nucleus. The effects of 5-HT receptor subtype agonists and antagonist were examined in polysynaptic neurons activated by stimulation of the ipsilateral abducens nucleus (IAN) antidromically, since these neurons are sensitive to 5-HT as shown in our previous study. Iontophoretic application of 5-HT and 8-hydroxy-2-(di-n-propylamino)tetrain (8-OH-DPAT), a selective 5-HT1A agonist, inhibited orthodromic spikes elicited by vestibular nerve stimulation in the majority of polysynaptic neurons activated by stimulation of ipsilateral IAN antidromically. There was a good correlation between the effects of 5-HT and 8-OH-DPAT. Iontophoretically applied 5-HT and 8-OH-DPAT also inhibited glutamate-induced firing in these neurons. Simultaneous application of 1-(2-methoxyphenyl)-4-[4-(2-phthalimido)butyl]piperazine (NAN-190), a 5-HT1A agonist/antagonist, significantly antagonized the 8-OH-DPAT-induced inhibition of glutamate-induced firing, although NAN-190 alone also caused weak suppression of glutamate-induced firing. Microiontophoretically applied 1-(3-chlorophenyl)piperazine (mCPP), a 5-HT1B agonist inhibited the orthodromic spike elicited by vestibular nerve stimulation and glutamate-induced firing in only a small number of the LVN neurons. 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), a 5-HT2 agonist, rarely affected these neurons. We postulate that postsynaptically located 5-HT1A receptors are mainly involved in the 5-HT-induced inhibition of polysynaptic neurons projecting in the region of the IAN.

8-Hydroxy-2-(di-n-propylamino)tetralin↗

3,3'-Iminodipropionitrile induces neurofilament accumulations in the perikarya of rat vestibular ganglion neurons.

Exposure of rats to 3,3'-iminodipropionitrile (IDPN) results in neurofilament (NF)-filled swellings in the proximal axons of a number of large neurons, including sensory neurons in the dorsal root ganglia (DRG) and motor neurons in the spinal cord. The present report describes the effects of acute and chronic IDPN exposure on the vestibular ganglion (VG) neurons as compared to those on the DRG neurons. In the VG, IDPN induced intra-perikaryal accumulation of morphologically and immunocytochemically identified NFs. In the DRG of the same treated animals, IDPN induced proximal axonal swelling but no perikaryal NF accumulations. We concluded that the VG neurons preferentially express the IDPN-induced NF pathology in their myelinated cell bodies. It is hypothesized that the NF pathology occurring after IDPN is preferentially expressed in myelinated structures.

Animals↗

Neural connections between embryonic stem cell-derived neurons and vestibular hair cells in vitro.

This study aimed to examine the potential of embryonic stem cell (ESC)-derived neural progenitors for restoration of the neural network in the peripheral vestibular system. Mouse ESC-derived neural progenitors were co-cultured with explants of vestibular sensory epithelia from neonatal mice. Histological analyses demonstrated that ESC-derived neurons substantially elongated their neurites towards vestibular hair cells, and attached to hair cells at the regions corresponding to the location of nerve endings in normal vestibular epithelia. Immunoreactivity for synaptophysin, a marker for synaptic vesicles, was present only in the cytoplasm of hair cells in sensory epithelia cultured alone, while the nerve endings of ESC-derived neurons attached to hair cells exhibited intense immunoreactivity for synaptophysin and some hair cells were moderately reactive in co-cultured specimens. The pattern of synaptophysin expression in co-cultured specimens was very similar to that observed in developing sensory epithelia, in which synaptic connections between hair cells and nerve endings are actively formed. These findings indicate that ESC-derived neurons have the potential to restore neural connections in the peripheral vestibular system.

Age Factors↗

Contribution of the vestibular primary neuron and the sensory cell to posture recovery.

The right vestibular nerve of the frog was cut peripherally to the vestibular ganglion. The frog sustained a tilting posture toward the right side. This tilting disappeared and the frog returned to normal posture. Based on this finding, the following experiments were done: In experiment I, the right vestibular nerve was again cut after posture recovery. The tilting angle was smaller and the recovery period shorter than after the 1st neurectomy. In experiment II, the left nerve was cut after posture recovery. The tilting angle toward the left was greater than that of the 1st neurectomy. In experiment III, the frog underwent bilateral neurectomy. A piece of the bone was inserted into the cut ends of the right nerve to inhibit nerve regeneration. The frog slowly developed tilting toward the right side. The above results indicate that both reactivation of the endorgan and the central compensatory mechanism play essential roles, for postural recovery after vestibular neurectomy.

Animals↗