Roles of glutamate receptor subtypes in the development of vestibular compensation after unilateral labyrinthectomy in the guinea pig

We investigated the roles of ionotropic glutamate receptor subtypes in the development and recovery of spontaneous nystagmus (SN) after unilateral labyrinthectomy (UL) in guinea pigs. When administered at 3 h after UL, N-methyl-D-aspartate (NMDA) and kainate (KA), which are NMDA and non-NMDA receptor agonists, respectively, increased the frequency of SN. The effect of KA was more potent than that of NMDA. In contrast to these agonists, MK-801 and CNQX decreased the frequency of SN. Although the administration of KA at 48 h after UL increased the frequency of SN, it did not exhibit any effects at 72 h after UL. MK-801 caused a recurrence of SN following administration at 48 and 72 h after UL. Neither NMDA nor CNQX exhibited any effects after administration at 48 or 72 h after UL. A newly synthesized compound, NC-1200, which has inhibitory action on the glutamate response, decreased the frequency of SN in a dose-dependent manner following administration at 3 h after UL, but did not exhibit any effects when administered at 48 and 72 h after UL. From these results, it was found that NMDA and non-NMDA receptors play important roles in the development of SN after UL, and that the NMDA receptor contributes to the development of ocular motor compensation.     

GABA<Subscript>A</Subscript> receptor subunit expression in the guinea pig vestibular nucleus complex during the development of vestibular compensation

The aim of this experiment was to investigate whether vestibular compensation following unilateral vestibular deafferentation (UVD) is associated with changes in the expression of GABA_A receptor subunits in the guinea pig vestibular nuclear complex (VNC) at 2, 10, and 30 h post-surgery. Using Western blotting, the ₁ and ₂ subunits (but not the ₂ subunit) were detected in the VNC of labyrinthine-intact animals. However, there were no significant differences in the protein expression of the ₁ and ₂ subunits within the ipsilateral or contralateral VNC at any time post-UVD compared to sham and anesthetic controls. Furthermore, UVD did not induce the expression of the ₂ protein. These results suggest that vestibular compensation in guinea pig, as in the rat, is not associated with changes in the protein levels of the GABA_A receptor subunits ₁, ₂, and ₂ in the VNC. However, a limitation of this study is that the Western blotting technique can detect only changes that are larger than 30% and therefore small changes cannot be excluded.     

Vestibular compensation without brainstem commissures in the guinea pig

The rapid recovery from the postural and ocular motor asymmetries produced by unilateral vestibular damage (vestibular compensation) has been presented as an example of plasticity in the central nervous system. A recent model (J. Neurophysiol., 51 (1984) 242-259) has identified the fibers joining the two vestibular nuclei in the brainstem (the vestibular commissures) as the site of the plastic changes. We report that in guinea pigs, compensation of postural symptoms still occurs after sectioning these commissural fibers. We suggest that a number of mechanisms may be responsible for vestibular compensation, including some which are independent of the vestibular commissures.     

Response of guinea pig vestibular nucleus neurons to clicks

Responses of single neurons in the vestibular nuclei to clicks were studied by extracellular recording in anaesthetised guinea pigs. Eighty-four neurons in the ipsilateral vestibular nuclei were activated with an average latency of 1.75±0.30 ms, which is about 0.9 ms longer than the mean latency of activation of click-sensitive vestibular afferents to intense clicks. The threshold of clicks for evoking the response of these neurons was around 70 dB above the auditory brainstem response threshold. Earlier studies have indicated that click-sensitive vestibular afferents are tilt-sensitive and likely to originate from saccular receptors, and in the present study nine of the click-sensitive vestibular nucleus neurons were tilt-sensitive, suggesting that these central neurons receive monosynaptic input from the corresponding saccular afferents. Recording sites were marked by means of iontophoretic injection of FCF green dye; they were located in the lateral portion of the descending vestibular nucleus and the caudal and ventral regions of the lateral vestibular nucleus.     

Responses of guinea pig primary vestibular neurons to clicks

Responses of single neurons in the vestibular nerve to high-intensity clicks were studied by extracellular recording in anaesthetised guinea pigs. One hundred and two neurons in the posterior division of the superior branch or in the inferior branch of the vestibular nerve were activated at short latency by intense clicks. The latency of activation was short (median 0.9 ms) and the threshold was high: the click intensity for evoking the response of these cells was around 60 dB above the auditory brainstem response threshold. Animals were tilted and rotated to identify physiologically the sensory region of the labyrinth from which the activated neurons originated. Seventeen neurons responded to static tilt as well as clicks. These results show that vestibular receptors, probably the otoliths, respond to clicks at intensities corresponding to those used in a new clinical test of the vestibulo-collic pathway.     

Motile responses of isolated guinea pig vestibular hair cells

Vestibular hair cells were isolated from the guinea pig vestibule by a micromechanical non-enzymatic procedure. Perfusion with 125 mM K+ solution induced irreversible slow shortening of the necks in 42.8% of the hair cells tested. Mechanical stimulation, creating a displacement of the hair bundle towards the kinocilium, induced either irreversible coiling or tilting of the neck of the cells, or reversible fast tilting of the cuticular plate (44.5% of tested cells). The response to the Ca2+ antagonist, Flunarizine, suggested that these movements were calcium-dependent. We propose several explanations of the physiological role of these mechanisms and discuss the possibility that fast tilting of the cuticular plate is a physiological movement involving the hair cells at the periphery of the vestibular receptors. The regulation of the vestibular message at the apex of type I hair cells is also considered.     

左氧氟沙星延长离体豚鼠乳头肌的动作电位时程

目的观察左氧氟沙星(LVFX)对豚鼠心肌动作电位的影响,并以司帕沙星(SPX)为阳性对照物比较二者对心肌的毒性作用。方法微电极记录豚鼠右心室乳头肌动作电位。结果刺激频率为1 Hz时,各浓度的SPX均显著性延长APD50和APD90,而LVFX浓度超过10μmol/L时明显延长APD50和APD90。结论LVFX和SPX均引起APD延长,前者效应较后者弱,仅在大剂量时有导致QT间期延长的潜在毒性。     

Non-typical K+-current in cesium-loaded guinea pig type I vestibular hair cell

Isolated guinea pig type I vestibular hair cells were voltage clamped at HP-110 mV in whole cell clamp configuration and depolarized up to +20 mV. Increasing depolarizations elicited large outward currents. These currents were replaced, in cesium-loaded cells, by inward/outward currents that reversed at membrane potentials between –55 and –30 mV. The reversal potential varied from cell to cell, and appeared to depend on the intracellular potassium cesium ratio. The current remaining in the presence of intracellular cesium was essentially due to a non-typical potassium conductance, which decreased in the presence of 4-AP and was blocked by 4-AP plus TEA. This current appeared as soon as the membrane was depolarized, showing the high potassium permeability of type I vestibular hair cells. A small part of this current was a strictly calcium inward current, sensitive to flunarizine, with a leakage component in the hyperpolarized state and a voltage component when the cell was depolarized.     

A phenomenon in vestibular compensation

The role of spinal afferentation from the lower half of the body in compensation of the sequelae of unilateral loss of vestibular function was studied in experiments on guinea pigs. Division of the spinal cord at the thoracic level under local anesthesia had no appreciable effect on the development of compensation after simultaneous or subsequent destruction of the labyrinth and did not disturb compensation in previously labyrinthectomized animals. Division of the spinal cord in labyrinthectomized animals under ether or chloroform anesthesia was accompanied by a sharp disturbance of compensation. These substances evoked a similar picture of decompensation in unilaterally labyrinthectomized animals with an intact spinal cord also. The results indicate that the disturbance of vestibular compensation discribed in the literature after division of the spinal cord under ether anesthesia is not the result of removal of spinal afferentation from the lower half of the body, but is due to the direct effect of inhalational anesthetics on compensation mechanisms.(Presented by Academician of the Academy of Medical Sciences of the USSR O. G. Gazenko.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 88, No. 7, pp. 21–23, July, 1979.     

Resonance of spike discharge modulation in neurons of the guinea pig medial vestibular nucleus

The modulation of action potential discharge rates is an important aspect of neuronal information processing. In these experiments, we have attempted to determine how effectively spike discharge modulation reflects changes in the membrane potential in central vestibular neurons. We have measured how their spike discharge rate was modulated by various current inputs to obtain neuronal transfer functions. Differences in the modulation of spiking rates were observed between neurons with a single, prominent after hyperpolarization (AHP, type A neurons) and cells with more complex AHPs (type B neurons). The spike discharge modulation amplitudes increased with the frequency of the current stimulus, which was quantitatively described by a neuronal model that showed a resonance peak >10 Hz. Modeling of the resonance peak required two putative potassium conductances whose properties had to be markedly dependent on the level of the membrane potential. At low frequencies (< or =0.4 Hz), the gain or magnitude functions of type A and B discharge rates were similar relative to the current input. However, resting input resistances obtained from the ratio of the membrane potential and current were lower in type B compared with type A cells, presumably due to a higher level of active potassium conductances at rest. The lower input resistance of type B neurons was compensated by a twofold greater sensitivity of their firing rate to changes in membrane potential, which suggests that synaptic inputs on their dendritic processes would be more efficacious. This increased sensitivity is also reflected in a greater ability of type B neurons to synchronize with low-amplitude sinusoidal current inputs, and in addition, their responses to steep slope ramp stimulation are enhanced over the more linear behavior of type A neurons. This behavior suggests that the type B MVNn are moderately tuned active filters that promote high-frequency responses and that type A neurons are like low-pass filters that are well suited for the resting tonic activity of the vestibular system. However, the more sensitive and phasic type B neurons contribute to both low- and high-frequency control as well as signal detection and would amplify the contribution of both irregular and regular primary afferents at high frequencies.     

Projection of primary vestibular afferent fibres to the cochlear nucleus in the guinea pig

After tracing the superior branch of the vestibular nerve and the macula sacculi by means of the neuronal tracers horseradish peroxidase (HRP) and wheat germ conjugated horseradish peroxidase (WGA-HRP), a conspicuous fibre bundle running into the cochlear nucleus could be observed. The HRP-labeled axons travel caudally through the descending vestibular nucleus, enter the cochlear nucleus at a level caudal to subgroup y and terminate at cells situated between the dorsal and posteroventral cochlear nucleus. Considering recent electrophysiological studies, it is reasonable to imply that these fibres are involved with the transduction of acoustic stimuli.     

Bone conducted vibration selectively activates irregular primary otolithic vestibular neurons in the guinea pig

The main objective of this study was to determine whether bone-conducted vibration (BCV) is equally effective in activating both semicircular canal and otolith afferents in the guinea pig or whether there is preferential activation of one of these classes of vestibular afferents. To answer this question a large number (346) of single primary vestibular neurons were recorded extracellularly in anesthetized guinea pigs and were identified by their location in the vestibular nerve and classed as regular or irregular on the basis of the variability of their spontaneous discharge. If a neuron responded to angular acceleration it was classed as a semicircular canal neuron, if it responded to maintained roll or pitch tilts it was classified as an otolith neuron. Each neuron was then tested by BCV stimuli—either clicks, continuous pure tones (200–1,500 Hz) or short tone bursts (500 Hz lasting 7 ms)—delivered by a B-71 clinical bone-conduction oscillator cemented to the guinea pig's skull. All stimulus intensities were referred to that animal's own auditory brainstem response (ABR) threshold to BCV clicks, and the maximum intensity used was within the animal's physiological range and was usually around 70 dB above BCV threshold. In addition two sensitive single axis linear accelerometers cemented to the skull gave absolute values of the stimulus acceleration in the rostro-caudal direction. The criterion for a neuron being classed as activated was an audible, stimulus-locked increase in firing rate (a 10% change was easily detectable) in response to the BCV stimulus. At the stimulus levels used in this study, semicircular canal neurons, both regular and irregular, were insensitive to BCV stimuli and very few responded: only nine of 189 semicircular canal neurons tested (4.7%) showed a detectable increase in firing in response to BCV stimuli up to the maximum 2 V peak-to-peak level we delivered to the B-71 oscillator (which produced a peak-to-peak skull acceleration of around 6–8 g and was usually around 60–70 dB above the animal's own ABR threshold for BCV clicks). Regular otolithic afferents likewise had a poor response; only 14 of 99 tested (14.1%) showed any increase in firing rate up to the maximum BCV stimulus level. However, most irregular otolithic afferents (82.8%) showed a clear increase in firing rate in response to BCV stimuli: of the 58 irregular otolith neurons tested, 48 were activated, with some being activated at very low intensities (only about 10 dB above the animal's ABR threshold to BCV clicks). Most of the activated otolith afferents were in the superior division of the vestibular nerve and were probably utricular afferents. That was confirmed by evidence using juxtacellular injection of neurobiotin near BCV activated neurons to trace their site of origin to the utricular macula. We conclude there is a very clear preference for irregular otolith afferents to be activated selectively by BCV stimuli at low stimulus levels and that BCV stimuli activate some utricular irregular afferent neurons. The BCV generates compressional and shear waves, which travel through the skull and constitute head accelerations, which are sufficient to stimulate the most sensitive otolithic receptor cells.     

Localization of pyroantimonate-precipitable calcium in the vestibular organs of the rat and guinea pig

Localization of pyroantimonate-precipitable calcium (Ca) was examined in the vestibular organs of the rat and the guinea pig. Precipitates were observed around the otoconia and in the otolithic membranes of the macula sacculi and macula utriculi. Although the deposition was heavier in the rat than in the guinea pig, the distribution of the Ca-pyroantimonate was similar in both species. Most otoconia in the dark cell area had precipitates around and inside the otoconia. Furthermore, there was some precipitation in the cupulae of the semicircular canals in both animals. The presence of Ca in the precipitates was confirmed by X-ray microanalysis and extraction with EGTA. Localization of the Ca-pyroantimonate around the otoconia in these animals agrees with localization of precipitates of endolymphatic crystals in the tree frog, which are known to grow mainly by accretion. Thus, the otoconia of the rat, and probably also of the guinea pig, can be assumed to grow mainly by accretion.     

Neuronal activity in the guinea pig medial vestibular nucleus in vitro following chronic unilateral labyrinthectomy

Unilateral labyrinthectomy (UL) causes ocular motor and postural disorders which disappear over time in a process of recovery known as vestibular compensation. Vestibular compensation is due to CNS plasticity which generates a partial recovery of resting activity in the vestibular nucleus ipsilateral to the UL, however the mechanism of this neural recovery is unknown. It has been suggested that other areas of the CNS may substitute non-vestibular sensory inputs for the missing labyrinthine input, thereby causing vestibular compensation. The present results show that resting activity can be recorded from medial vestibular nucleus (MVN) neurons in vitro, in brainstem slices from guinea pigs which have compensated for an ipsilateral UL. This result suggests that MVN neurons are capable of generating resting activity without inputs from many other CNS areas. Perfusion with high Mg2+ solution did not abolish resting activity in most cases, suggesting that part of the resting activity may be generated spontaneously by the neurons, possibly through changes in the electrical excitability of the cell membrane.     

Clinical anesthesia causes permanent damage to the fetal guinea pig brain

Exposure of the immature brain to general anesthesia is common. The safety of this practice has recently been challenged in view of evidence that general anesthetics can damage developing mammalian neurons. Initial reports on immature rats raised criticism regarding the possibly unique vulnerability of this species, short duration of their brain development and a lack of close monitoring of nutritional and cardiopulmonary homeostasis during anesthesia. Therefore, we studied the neurotoxic effects of anesthesia in guinea pigs, whose brain development is longer and is mostly a prenatal phenomenon, so that anesthesia-induced neurotoxicity studies of the fetal brain can be performed by anesthetizing pregnant female pigs. Because of their large size, these animals made invasive monitoring of maternal and, indirectly, fetal well-being technically feasible. Despite adequate maintenance of maternal homeostasis, a single short maternal exposure to isoflurane, whether alone or with nitrous oxide and/or midazolam at the peak of fetal synaptogenesis, induced severe neuroapoptosis in the fetal guinea pig brain. As detected early in post-natal life, this resulted in the loss of many neurons from vulnerable brain regions, demonstrating that anesthesia-induced neuroapoptosis can cause permanent brain damage.     

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.     

Calretinin immunoreactivity in chinchilla and guinea pig vestibular end organs characterizes the calyx unit subpopulation

Summary Immunohistochemical investigations with calretinin, a neuronal calcium binding protein, were made in the vestibular end organs of five guinea pigs and one chinchilla. A specific pattern of immunoreactivity of afferent nerve fibers was found. Immunostaining was restricted to thick fibers innervating the apex of the cristae or the striola of the utricular macula. A study of serial sections revealed that the stained afferents gave rise to calyx endings, but not to collaterals containing bouton endings. The results are consistent with the conclusion that, of the three classes of fibers defined by Fernendez et al. (1988, 1990), only calyx units are calretinin immunoreactive. A count of the number of labelled fibers in the chinchilla crista suggests that the entire population of calyx units is immunoreactive. The conclusion is surprising since the physiology of calyx units does not differ qualitatively from that of other afferents (Baird et al. 1988; Goldberg et al. 1990). The presence of this protein in the calyx neurons may be related to specific postsynaptic functions of this type of afferents.     

Thioperamide delays vestibular compensation in goldfish

Unilateral lesion of the vestibular system induces posturo-locomotor deficits that are compensated for with time. Drug therapy is currently used to improve the recovery process and to facilitate vestibular compensation. We investigated the effects of thioperamide on functional recovery after unilateral labyrinthectomy in Carassius auratus. Approximately 24h after surgery, the animals were injected intraperitoneally with thioperamide (15 mg/kg) and saline (1.5 ml/kg). The injections were repeated daily for a total of 15 consecutive days. The substances were administered in a volume of 1.5 ml/kg body weight. Another group, which served as a non-lesion control, did not receive unilateral labyrinthectomy or system injections. Animals treated with saline presented a compensatory decrease in body tilt on the 7th day, while the animals treated with thioperamide presented a decrease in body tilt from the 13th day, suggesting a delay in the functional recovery process. These results suggest that an increase in cerebral histamine levels impairs vestibular compensation in goldfish.