Tonic activity of rat medial vestibular nucleus neurones in vitro and its inhibition by GABA

Summary The spontaneous discharge of 48 medial vestibular nucleus (MVN) neurones was recorded extracellularly in horizontal slices of the rat brainstem in vitro. The mean tonic rate of discharge was 17.1±8.2 imp/s, similar to that observed by others in transverse (coronal) slices of the rat and guinea pig MVN. The tonic rate of discharge of individual MVN cells either increased or decreased after synaptic blockade in low Ca²⁺ media, suggesting that ongoing synaptic activity has an important influence on the spontaneous activity of MVN cells in vitro. However the persistence of tonic activity after synaptic blockade indicates that an intrinsic, pacemaker-like mechanism is involved in the generation of the tonic activity. GABA, muscimol, baclofen and 3-APA inhibited the tonic activity of all MVN cells tested. Bicuculline antagonised, and picrotoxin blocked, the inhibitory responses to muscimol, but the effects of GABA were only partially blocked in 50 M picrotoxin. The effects of baclofen and 3-APA persisted in low Ca²⁺ media, and were antagonised by saclofen and phaclofen. Picrotoxin-resistant responses to GABA persisted in low Ca²⁺ media, and were also antagonised by saclofen. These results suggest that the inhibitory control of MVN neurones by GABA involves both the GABA_A and GABA_B subtypes of GABA receptor. GABA_B receptors appear to be distributed both pre- and post-synaptically in the rat MVN. The possible significance of the intrinsic, tonic activity of MVN cells in normal vestibular function and in vestibular compensation, and the effects of GABA, are discussed.     

Age-related neuronal loss in the cochlea is not delayed by synaptic modulation

Age-related synaptic change is associated with the functional decline of the nervous system. It is unknown whether this synaptic change is the cause or the consequence of neuronal cell loss. We have addressed this question by examining mice genetically engineered to over- or underexpress neuregulin-1 (NRG1), a direct modulator of synaptic transmission. Transgenic mice overexpressing NRG1 in spiral ganglion neurons (SGNs) showed improvements in hearing thresholds, whereas NRG1 −/+ mice show a complementary worsening of thresholds. However, no significant change in age-related loss of SGNs in either NRG1 −/+ mice or mice overexpressing NRG1 was observed, while a negative association between NRG1 expression level and survival of inner hair cells during aging was observed. Subsequent studies provided evidence that modulating NRG1 levels changes synaptic transmission between SGNs and hair cells. One of the most dramatic examples of this was the reversal of lower hearing thresholds by “turning-off” NRG1 overexpression. These data demonstrate for the first time that synaptic modulation is unable to prevent age-related neuronal loss in the cochlea.     

The dynamic sensitivity of pyramidal tract neurones is influenced by previous activity

Summary The dynamic sensitivity of Pyramidal Tract (PT) neurones has been examined in silent cells and after pre-activation with an action potential evoked at different intervals before the discharge onset. In the resting conditions the dynamic sensitivity increases after the first interspike interval of the repetitive discharge. When a conditioning spike precedes the discharge onset by 50 ms (i.e. of a time sufficient for its afterhyperpolarization, AHP, to fade out) the dynamic sensitivity becomes uniform throughout the ramp. This linearizing effect progressively declines when the conditioning interval is increased to 100 and 150 ms and is attributed to the depression induced by the conditioning spike on the AHP of the first spike of the ramp discharge. In natural conditions, the effect would be entirely developed when the neurones fire at their minimal discharge rate (1/AHP duration).     

Reappearance of electroencephalogram slow waves in extended sleep with delayed bedtime

A three-part study using prolonged nights of sleep was undertaken to verify Broughton's hypothesis of an approximate 12-h ultradrian rhythm of human slow wave sleep (SWS). Part I consisted of 2 8-h adaptation nights followed by a prolonged 15-h night of sleep with bedtime at midnight. A significant return of SWS occurred 12 h and 32 min after the first appearance of SWS. In part II, after 1 adaptation night, subjects were asked to sleep for 15 h but bedtime was delayed until 0400 h. A two-peak return of SWS was observed with a first significant return at 1228 h and a second significant return at 1745 h (i.e., 13 h and 32 min after the first appearance of SWS). In part III, bedtime was again delayed to 0400 h; but subjects were given 3 nights to adapt before the 15 h extended sleep. A single significant return of SWS was then observed at 1656 h, i.e., 12 h and 24 min after the first appearance with no peaks around 1200 h, thus exhibiting the same pattern as in part I. These results suggest that the return of SWS seen normally between 1200 and 1500 h is relatively well entrenched since it remained present in the extended night following sudden bedtime delay. It appears, moreover, that SWS does follow a bimodal 12-h rhythm, which is seen immediately upon extended delayed sleep and can be fully phase-shifted with habituation.     

The delayed rectifier,Ikh, is the major conductance in type i vestibular hair cells across vestibular end organs

Hair cells were dissociated from the semicircular canal, utricle, lagena and saccule of white king pigeons. Type I hair cells were identified morphologically based on the ratios of neck width to cuticular plate width (NPR < 0.72) as well as neck width to cell body width (NBR < 0.64). The perforated patch variant of the whole-cell recording technique was used to measure electrical properties from type I hair cells. In voltage-clamp, the membrane properties of all identified type I cells were dominated by a predominantly outward potassium current, previously characterized in semicircular canal as I _KI. Zero-current potential, activation, deactivation, slope conductance, pharmacologic and steady-state properties of the complex currents were not statistically different between type I hair cells of different vestibular end organs. The voltage dependence causes a significant proportion of this conductance to be active about the cell’s zero-current potential. The first report of the whole-cell activation kinetics of the conductance is presented, showing a voltage dependence that could be best fit by an equation for a single exponential. Results presented here are the first data from pigeon dissociated type I hair cells from utricle, saccule and lagena suggesting that the basolateral conductances of a morphologically identified population of type I hair cells are conserved between functionally different vestibular end organs; the major conductance being a delayed rectifier characterized previously in semicircular canal hair cells as I _KI. Received: 19 July 1995/Received after revision: 19 October 1995 Accepted: 25 October 1995     

Identification of the tracheal and laryngeal afferent neurones mediating cough in anaesthetized guinea-pigs

We have identified the tracheal and laryngeal afferent nerves regulating cough in anaesthetized guinea-pigs. Cough was evoked by electrical or mechanical stimulation of the tracheal or laryngeal mucosa, or by citric acid applied topically to the trachea or larynx. By contrast, neither capsaicin nor bradykinin challenges to the trachea or larynx evoked cough. Bradykinin and histamine administered intravenously also failed to evoke cough. Electrophysiological studies revealed that the majority of capsaicin-sensitive afferent neurones (both Aδ- and C-fibres) innervating the rostral trachea and larynx have their cell bodies in the jugular ganglia and project to the airways via the superior laryngeal nerves. Capsaicin-insensitive afferent neurones with cell bodies in the nodose ganglia projected to the rostral trachea and larynx via the recurrent laryngeal nerves. Severing the recurrent nerves abolished coughing evoked from the trachea and larynx whereas severing the superior laryngeal nerves was without effect on coughing. The data indicate that the tracheal and laryngeal afferent neurones regulating cough are polymodal Aδ-fibres that arise from the nodose ganglia. These afferent neurones are activated by punctate mechanical stimulation and acid but are unresponsive to capsaicin, bradykinin, smooth muscle contraction, longitudinal or transverse stretching of the airways, or distension. Comparing these physiological properties with those of intrapulmonary mechanoreceptors indicates that the afferent neurones mediating cough are quite distinct from the well-defined rapidly and slowly adapting stretch receptors innervating the airways and lungs. We propose that these airway afferent neurones represent a distinct subtype and that their primary function is regulation of the cough reflex.     

Electrical stimulation of vestibular nuclei: Effects on light-evoked activity of lateral geniculate nucleus neurones

Summary The light-evoked activity of single LGN neurones was recorded from light-adapted, anaesthetized cats. The neuronal responses to discrete illumination of the receptive field centre or periphery were compared to those obtained when the same photic stimulus was slightly preceded by electrical stimulation of the vestibular nuclei.It was found that the light-evoked responses of 82% of the LGN neurones studied were significantly affected by the vestibular stimulus. Both facilitatory and inhibitory effects were observed. There were no laterality effects. Similar modulation of LGN light-evoked activity was observed following stimulation of the mesencephalic reticular formation.Steps taken to eliminate various sources of experimental error are described. The above evidence suggests that the LGN is not a mere relay station but rather a structure where sensory interaction occurs.     

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.     

Chondrosarcoma is not characterized by detectable telomerase activity

Reactivation of telomerase, an enzyme which elongates human telomeres, is associated with cell immortilization. In approximately 90% of malignant tumours telomerase activity can be demonstrated, whereas in benign tumours it is mostly absent. Chondrosarcomas are relatively rare malignant cartilaginous neoplasms. A small number of chondrosarcomas located centrally in bone arise secondarily to an enchondroma, while the majority of chondrosarcomas developing from the surface arise within the cartilage cap of an osteochondroma. The histological distinction between a benign lesion and low-grade chondrosarcoma is generally considered difficult. To investigate whether the progression towards chondrosarcoma is characterized by reactivation of telomerase activity, this study determined telomerase activity in ten enchondromas, five osteochondromas, and 37 chondrosarcomas using the TRAP assay. In all tumour samples except one, telomerase activity was absent. By adding tumour lysates to the positive control, an increasing inhibition of telomerase activity was found with an increasing chondroid matrix, suggesting that it may contain inhibitory factors. Inhibition due to endogenous RNAse or Taq-polymerase inhibitors was excluded. The lack of detectable telomerase activity in the high-grade component of a dedifferentiated chondrosarcoma without matrix favours the possibility that telomerase is truly absent. Either its true absence or inhibitory effects disabling telomerase detection exclude the telomerase TRAP assay as a diagnostic tool in the differential diagnosis of benign and low-grade malignant cartilaginous tumours.     

Erythrocyte acetylcholinesterase activity is not affected by adenovirus-induced haemagglutination

No correlation was found between adenovirus-induced haemagglutination and AChE activity of human RBCs as ascertained with three different types of virus. The haemagglutination titre of the adenoviruses did not decrease when RBCs with AChE inhibited by dichlorvos or eserine were used. The receptors involved in active haemagglutination by adenoviruses thus seem to differ from those acting in passive haemagglutination. The AChE inactivating effect of substances such as tannic acid and glutaraldehyde, among others, might be due to a non-specific damage of the RBC membrane.     

Configural processing of other-race faces is delayed but not decreased

The processing of other-race faces has been suggested to differ from own-race face processing with regard to the extent to which configural and/or holistic information is taken into account. We aimed at investigating the underlying mechanisms more precisely by applying event-related potentials (ERP). We presented upright and inverted own-race, other-race, and other-species faces (apes), as well as non-facial control stimuli (houses), and analysed the effects of stimulus type and inversion on the N170. Peak latencies for same-race, other-race, other-species, and non-face stimuli gradually increased, and inversion of all types of face stimuli led to an additional delay. Importantly, the two factors did not interact for face stimuli. Inversion increased N170 amplitudes for own- and other-race faces only. We conclude that early perceptual mechanisms of face processing are not qualitatively different for own- and other-race faces. Instead, the same underlying processes appear to work less efficiently for other-race faces.     

Spontaneous synaptic activity is primarily GABAergic in vestibular nucleus neurons of the chick embryo

The principal cells of the chick tangential nucleus are vestibular nucleus neurons participating in the vestibular reflexes. In 16-day embryos, the application of glutamate receptor antagonists abolished the postsynaptic responses generated on vestibular-nerve stimulation, but spontaneous synaptic activity was largely unaffected. Here, spontaneous synaptic activity was characterized in principal cells from brain slices at E16 using whole cell voltage-clamp recordings. With KCl electrodes, the frequency of spontaneous inward currents was 3.1 Hz at -60 mV, and the reversal potential was +4 mV. Cs-gluconate pipette solution allowed the discrimination of glycine/GABA(A) versus glutamate receptor-mediated events according to their different reversal potentials. The ratio for spontaneous excitatory to inhibitory events was about 1:4. Seventy-four percent of the outward events were GABA(A), whereas 26% were glycine receptor-mediated events. Both pre- and postsynaptic GABA(B) receptor effects were shown, with presynaptic GABA(B) receptors inhibiting 40% of spontaneous excitatory postsynaptic currents (sEPSCs) and 53% of spontaneous inhibitory postsynaptic currents (sIPSCs). With TTX, the frequency decreased approximately 50% for EPSCs and 23% for IPSCs. These data indicate that the spontaneous synaptic activity recorded in the principal cells at E16 is primarily inhibitory, action potential-independent, and based on the activation of GABA(A) receptors that can be modulated by presynaptic GABA(B) receptors.     

Sodium-potassium-ATPase activity is influenced by ethnic origin and not by obesity

Previous investigations have suggested that red-cell ouabain binding (an indirect measure of sodium-potassium-ATPase activity) is lower in severely obese patients than in normal controls. We now confirm that ouabain binding measures sodium-potassium-ATPase activity, and we demonstrate that the level of this activity is genetically determined. The activity of this enzyme differs in various ethnic and racial groups, relatively high levels being encountered in non-Jewish white subjects, particularly those with some Scandinavian ancestry. On the other hand, black, Asian, and Jewish white subjects have lower sodium-potassium-ATPase activity. In contrast, no difference was found in red-cell sodium-potassium-ATPase activity between severely obese and normal persons, nor could we confirm a putative effect of food intake on the level of the red-cell enzyme. We suggest that in most earlier studies in which differences were found between normal and severely obese persons, those differences could have been due to differences in the ethnic origins of the obese and control populations.     

An in situ hybridization and immunofluorescence study of glycinergic receptors and gephyrin in the vestibular nuclei of the intact and unilaterally labyrinthectomized rat

We investigated whether the expression of glycinergic receptor (GLYR) subunits of gephyrin and of their mRNAs in the medial vestibular nuclei are affected following unilateral labyrinthectomy. Specific radioactive oligonucleotide probes recognizing the sequences encoding 1–3 and ß subunits of GLYR and the anchoring protein gephyrin were used to probe sections of vestibular nuclei. Signals in these in situ hybridization experiments were detected with film or by emulsion photography. Animals were killed at various times following the lesion: 5 h, 1, 3, 8, 30 and 60 days. Specific monoclonal GLYR and gephyrin antibodies were also used to determine GLYR and gephyrin immunoreactivity in control and operated rats (5 h, 1, 3 and 8 days post-lesion). In normal animals, several brainstem regions including the lateral, medial, superior and inferior vestibular nuclei contained mRNAs for gephyrin and the 1 and subunits of GLYR, and expressed the GLYR and gephyrin polypeptides. In unilaterally labyrinthectomized rats, no asymmetry was detected on autoradiographs between the two medial vestibular nuclei with any of the oligonucleotide probes used, or at any time following the lesion. No difference in the immunofluorescence staining was observed between the intact and deafferented medial vestibular nuclei of lesioned animals or between the vestibular nuclei of lesioned and controls rats. Thus, deafferentation of the vestibular nuclei did not affect the expression of gephyrin, of the various GLYR subunits, or of their mRNAs in the deafferented and intact medial vestibular nuclei. It is therefore unlikely that GLYR and gephyrin modulation contribute significantly to the recovery of the resting discharge of the deafferented medial vestibular neurons and consequently to the restoration of a normal posture and eye position.Abbreviations AMPA -amino-3-hydroxy-5-methylisoxazole-4-propionic acid - AF Arithmetic fluorescence - CNS Central nervous system - DTT Dithiothreitol - EGTA Ethylene glycol tetraacetic acid - GABA -aminobutyric acid - GLY Glycine - GLYR Glycine receptors - MVN Medial vestibular nucleus - MVNn Medial vestibular nucleus neurons - OD Optical density - PBS Phosphate buffered saline - mRNA Messenger ribonucleic acid - MVNn Medial vestibular nuclei neurons - NMDA N-methyl-D-aspartate - rAF Relative arithmetic fluorescence - rOD Relative optical density - tRNA Transfer ribonucleic acid - SD Standard deviation - SSC Standard saline citrate - VNn Vestibular nuclei neurons     

Light-induced activity in the activity box is not aversively motivated and does not show between-trial habituation

The induction of behaviour by sensory stimuli, i.e. sensorimotor stimulation, is a fundamental aspect of behaviour. Recently, it was found that the presentation of white-light stimuli to a rat in an activity box reliably induces locomotor activity, and, thus, may be able to serve as a paradigm to measure basal, non-aversively motivated sensorimotor processing. However, light can be an aversive stimulus to a rat. In order to test if there is a stressful component in light-induced activity, a retreat-box was introduced into the test-apparatus in experiment 1, so that the animals had the opportunity to escape the light stimuli. It was found, that light-induced activity was also shown, when a retreat-box was available in the activity box, and that light-stimulation did not lead to an increase of entries into or the time spent in the retreat box. Experiment 2 examines the stability of the response to light over trials. Three light-induced activity test-trials were conducted with one day between each test. There was no effect of repeated testing on light-induced activity, which was evident during each of the three test-sessions. It is concluded that stress/anxiety does not significantly contribute to the increase of locomotor behaviour induced by light stimulation under the present conditions. Thus, the paradigm appears to involve a non-aversively motivated behavioural response. Furthermore, light-induced activity did not habituate over at least three test trials, and may, therefore, serve for repeated testing.     

Morphine, but not atropine, blocks nociceptor-driven activity in rat dorsal hippocampal neurones

Neurones in the CA1 region of the dorsal hippocampus of urethane-anaesthetized rats were recorded extracellularly with microelectrodes, and tested for their response to non-noxious and noxious peripheral stimuli. None of the cells responded to non-noxious stimuli but 91 of 216 cells were excited by noxious stimuli. Morphine (5.0 mg/kg i.v.) increased the background firing rate in 7 of 10 cells tested and blocked the response to noxious heating of the tail in all cases. Naloxone (0.5 mg/kg i.v.) reversed these effects of morphine. Atropine (0.1 plus 0.2 mg/kg i.v.), in another group of animals, produced only transient changes of an inconsistent nature in 7 cells tested.     

Development of action potentials and apamin-sensitive after-potentials in mouse vestibular nucleus neurones

The postnatal maturation of medial vestibular nucleus (MVN) neurones was examined in slices of the dorsal brainstem prepared from balb/c mice at specific stages during the first postnatal month. Using spike-shape averaging to analyse the intracellularly recorded action potentials and after-hyperpolarisations (AHPs) in each cell, all the MVN neurones recorded in the young adult (postnatal day 30; P30) mouse were shown to have either a single deep AHP (type A cells), or an early fast and a delayed slow AHP (type B cells). The relative proportions of the two subtypes were similar to those in the young adult rat. At P5, all the MVN cells recorded showed immature forms of either the type A or the type B action potential shape. Immature type A cells had broad spontaneous spikes, and the characteristic single AHP was small in amplitude. Immature type B cells had somewhat narrower spontaneous spikes that were followed by a delayed, apamin-sensitive AHP. The delayed AHP was separated from the repolarisation phase of the spike by a period of isopotentiality. Over the period P10–P15, the mean resting potentials of the MVN cells became more negative, their action potential fall-times became shorter, the single AHP in type A cells became deeper, and the early fast AHP appeared in type B cells. Until P15 cells of varying degrees of electrophysiological maturity were found in the MVN but by P30 all MVN cells recorded were typical adult type A or type B cells. Exposure to the selective blocker of SK-type Ca-activated K channels, apamin (0.3 μM), induced depolarising plateaux and burst firing in immature type B cells at rest. The duration of the apamin-induced bursts and the spike frequency during the bursts were reduced but not abolished after blockade of Ca channels in Ca-free artificial cerebrospinal fluid containing Cd²⁺. By contrast, in mature type B cells at rest apamin selectively abolished the delayed slow AHP but did not induce bursting activity. Apamin had no effect on the action potential shape of immature type A cells. These data show that the apamin-sensitive I _AHP is one of the first ionic conductances to appear in type B cells, and that it plays an important role in regulating the intrinsic rhythmicity and excitability of these cells. Received: 19 November 1996 / Accepted: 30 June 1997