Rhesus monkey vestibular cortex: a bimodal primary projection field.

Single units in the rhesus monkey (Macaca mulata) cortex responded to both vestibular and proprioceptive somatosensory stimuli. This bimodal response characteristic is unlike the modality specificity noted for other primary sensory fields. The vestibular field is located, contrary to previous opinion, within a distinct cytoarchitectonic area outside of area 2.

Human auditory steady state responses to binaural and monaural beats.

OBJECTIVE: Binaural beat sensations depend upon a central combination of two different temporally encoded tones, separately presented to the two ears. We tested the feasibility to record an auditory steady state evoked response (ASSR) at the binaural beat frequency in order to find a measure for temporal coding of sound in the human EEG. METHODS: We stimulated each ear with a distinct tone, both differing in frequency by 40Hz, to record a binaural beat ASSR. As control, we evoked a beat ASSR in response to both tones in the same ear. We band-pass filtered the EEG at 40Hz, averaged with respect to stimulus onset and compared ASSR amplitudes and phases, extracted from a sinusoidal non-linear regression fit to a 40Hz period average. RESULTS: A 40Hz binaural beat ASSR was evoked at a low mean stimulus frequency (400Hz) but became undetectable beyond 3kHz. Its amplitude was smaller than that of the acoustic beat ASSR, which was evoked at low and high frequencies. Both ASSR types had maxima at fronto-central leads and displayed a fronto-occipital phase delay of several ms. CONCLUSIONS: The dependence of the 40Hz binaural beat ASSR on stimuli at low, temporally coded tone frequencies suggests that it may objectively assess temporal sound coding ability. The phase shift across the electrode array is evidence for more than one origin of the 40Hz oscillations. SIGNIFICANCE: The binaural beat ASSR is an evoked response, with novel diagnostic potential, to a signal that is not present in the stimulus, but generated within the brain.

The primary vestibular projection to the cerebellar cortex in the pigeon (Columba livia).

The cerebellar cortex of the pigeon receiving direct vestibular afferents was delineated by anterograde transport of [3H]-amino acids injected into the vestibular nerve. Labelled mossy fiber rosettes in the granular layer were concentrated in lobule X (nodulus) and to a lesser extent, in the ventral portion of lobule IXd (uvula and paraflocculus). A few solitary labelled rosettes were also found in more dorsal portions of lobule IX, as well as in the anterior lobe between lobule II and IV. The lingula remained unlabelled. Discrete injections of [3H]-leucine into the cristae of each of the three semicircular canals or the utricular macula yielded a similar distribution of fewer labelled rosettes. A few primary mossy fiber terminals labelled after cochlear injections are attributed to afferents from the lagenar macula. Since effective diffusion of label from the injection site was excluded by controls, it is concluded that projection of individual canal and macula nerves to the vestibulocerebellar cortex is not topographically separated. It is proposed that this extensive convergence of various afferents is required by the cerebellum to compute precise and directionally specific control signals during head rotation in all conceivable planes.

Efferent vestibular neurons: a study employing retrograde tracer methods in the pigeon (Columba livia).

Efferent neurons innervating the vestibular labyrinth and cochlea of the pigeon have been identified by means of a variety of retrograde tracers: [3H]-adenosine (Ad), horseradish peroxidase (HRP), Evan's Blue (EB) and Bisbenzimide (Bb). Discrete injections into individual cristae ampullares of the semicircular canals, into the macula utriculi, or into several of these end organs resulted in similar patterns of neuronal labelling. Efferent vestibular neurons were always found within a small portion of the nucleus reticularis pontis caudalis (RP), ventrolateral to the abducens nucleus on both sides. No systemic difference in the locations of labelled cells was found following injection into different sensory epithelia. Cell counts following injections into individual cristae did not differ significantly from those following injections into all three cristae. The injections into all cristae in both labyrinths yielded cell counts that were much lower than twice the number of cells labelled by injections into the three cristae on one side only. When HRP was injected into the right lateral canal crista and Ad into the right posterior canal crista, a high proportion of neurons was labelled with both compounds (61% of the HRP-labelled cells and 67% of the Ad-labelled cells). Injections of EB into all three cristae on the right side and Bb into all three cristae on the left side produced a smaller percentage of doubly labelled cells (10% of the EB-labelled cells and 6% of the Bb-labelled cells). It is concluded, therefore, that there is a considerable degree of collateralization within one labyrinth. Fewer collaterals of efferent neurons are directed to both labyrinths. Since each semicircular canal represents head rotation in one direction and one plane, it is unlikely that efferents which contact several different movement sensors can provide sensory motor control that is specific for directions and planes of head movements. Control injections of these tracers into the cochlea yielded labelled cells in a different reticular structure, the nucleus reticularis paragigantocellularis lateralis (Pgc), on both sides, as well as in the RP. It is proposed that the Pgc cells represent cochlear efferents, while the RP neurons are related to the macula lagenae, an otolithic organ of balance in the pigeon.

Modulation of bursts and high-threshold calcium spikes in neurons of rat auditory thalamus.

Neurons in the ventral partition of the medial geniculate body are able to fire high-threshold Ca2+-spikes. The neurons normally discharge such spikes on low-threshold Ca2+-spikes after the action potentials of a burst. We studied membrane mechanisms that regulate the discharge of high-threshold Ca2+-spikes, using whole-cell recording techniques in a slice preparation of rat thalamus. A subthreshold (persistent) Na+-conductance amplified depolarizing inputs, enhancing membrane excitability in the tonic firing mode and amplifying the low-threshold Ca2+-spike in the burst firing mode. Application of tetrodotoxin blocked the amplification and high-threshold Ca2+-spike firing. A slowly inactivating K+ conductance, sensitive to blockade with 4-aminopyridine (50-100 microM), but not tetraethylammonium (2-10 mM), appeared to suppress excitability and high-threshold Ca2+-spike firing. Application of 4-aminopyridine increased the low-threshold Ca2+-spike and the number of action potentials in the burst, and led to a conversion of the superimposed high-threshold Ca2+-spike into a plateau potential. Application of the Ca2+-channel blocker Cd2+ (50 microM), reduced or eliminated this plateau potential. The tetrodotoxin sensitive, persistent Na+-conductance also sustained plateau potentials, triggered after 4-aminopyridine application on depolarization by current pulses. Our results suggest that high-threshold Ca2+-spike firing, and a short-term influx of Ca2+, are regulated by a balance of voltage-dependent conductances. Normally, a slowly inactivating A-type K+-conductance may reduce high-threshold Ca2+-spike firing and shorten high-threshold Ca2+-spike duration. A persistent Na+-conductance promotes coupling of the low-threshold Ca2+-spike to a high-threshold Ca2+-spike. Thus, the activation of both voltage-dependent conductances would affect Ca2+ influx into ventral medial geniculate neurons. This would alter the quality of the different signals transmitted in the thalamocortical system during wakefulness, sleep and pathological states.

Intrinsic response properties of bursting neurons in the nucleus principalis trigemini of the gerbil.

In trigeminal neurons, the spike rate, modulated by input parameters, may serve as a code for sensory information. We investigated intrinsic response properties that affect rate coding in neurons of nucleus principalis trigemini (young gerbils). Using the whole-cell recording technique and neurobiotin staining in slices, we found bursting behaviour in approximately 50% of the neurons. These neurons fired spike bursts, spontaneously, as well as at the onset of depolarizing, and offset of hyperpolarizing, current pulses. The spike rate within an initial burst was independent of stimulus strength, in contrast to single spike firing that occurred later in the response to current pulse injection. The spikes within a burst were superimposed on slow depolarizing humps. Under favourable conditions, these led to "plateau potentials", that lasted for hundreds of milliseconds at membrane potentials near approximately -20 mV. Occasionally, plateau potentials were spontaneous or evoked under control conditions: usually, they were evoked by current pulse injection during blockade of Ca2+ influx with Co2+ or Cd2+ in Ca(2+)-free extracellular media, or during blockade of K+ currents with tetraethylammonium. The plateau potentials recorded during internal Cs+ (132.5 mM) substitution of K+ had more positive amplitudes (near +20 mV). Despite relatively stable depolarization levels, the plateau potentials decreased in duration and decayed in amplitude during application of tetrodotoxin (0.6-1.8 nM). Higher tetrodotoxin concentrations (5-60 nM) eliminated the plateau potentials despite well-maintained, fast action potentials. A reduction of external [Na+] reduced the amplitudes of the spikes and plateau potentials. A hyperpolarization of long duration (> 3 s) followed a plateau potential, or a depolarizing response that was subthreshold for plateau generation. Tetrodotoxin application blocked this after-effect. We suggest that a persistent Na+ influx is a major contributor to the bursts and plateau potentials and that it mediates the hyperpolarization. Depending on Ca2+ influx, K+ conductances may regulate the amplitudes of these long-lasting depolarizations. A Ca2+ conductance, blockable by Ni2+, may support burst initiation in these neurons. In very young animals (P2-P9), we found only non-bursting neurons. Both bursting and non-bursting neurons with elongated dendritic fields showed inward rectification on hyperpolarization. The bursts in nucleus principalis trigemini neurons emphasize the onsets of stimulus transients, at the expense of using firing rate as a sensory code. Our studies describe neurons with a surprising ability to distort sensory signals, transforming depolarizing inputs into bursts of spikes by virtue of a Na(+)-conductance activation. The principal trigeminal nucleus also contains neurons with tonic firing ability, compatible with simple rate coding.

Postnatal development of signal generation in auditory thalamic neurons.

Using whole cell recording techniques, we distinguished immature from mature stages of development in auditory thalamic neurons of rats at ages P5 to P21. We compared voltage responses to injected currents and firing patterns of neurons in ventral partition of medial geniculate body (MGBv) in slices. Resting potential, input resistance and membrane time constant diminished to mature values between P5 and P14. Responses of young neurons to hyperpolarizing pulses showed delayed inward rectification; after P13, this was obscured by a rapid onset of another inward rectifier. All neurons possessed tetrodotoxin (TTX)-sensitive, depolarization-activated rectification, implying persistent Na+-current involvement. Despite a slightly higher voltage threshold for spiking, the current threshold was lower in younger neurons. Young neurons fired a short latency spike with afterhyperpolarization whereas older neurons exhibited a slow ramplike depolarization before tonic firing. Large currents caused continuous firing in all neurons. Before day P13, a high threshold Ca2+ spike (HTS) often was appended to action potentials. The low threshold Ca2+-spike (LTS) was too small in amplitude to evoke action potentials before P11 but produced a single spike at P12 and P13 and burst firing with HTS after P13. MGBv neurons have mature properties after P14, relevant for reactions to sound and the oscillations of slow-wave sleep.

Intonation of musical intervals by musical intervals by deaf subjects stimulated with single bipolar cochlear implant electrodes.

Some subjects with cochlear implants have been shown to associate electrical stimulus pulse rates with the pitches of musical tones. In order to clarify the role of these pitch sensations in a musical context, the present investigation examined the intonation accuracy achieved by implant subjects when adjusting pulse rates in the reconstruction of musical intervals. Using a method of adjustment, the subjects altered a variable pulse rate, relative to a fixed reference rate, on one electrode, in the tuning of musical intervals abstracted from familiar melodies. At low pulse rates, subjects generally tuned the intervals to the same frequency ratios which define tonal musical intervals in normal-hearing listeners, with error margins comparable to musically untrained subjects. Two subjects were, in addition, able to transpose these melodic intervals from a standard reference pulse rate to higher and lower reference rates (reference and target pulse rates with geometric means of the intervals ranging from 81 to 466 pulses/s). Generally, the intervals were adjusted on a ratio scale, according to the same frequency ratios which define analogous acoustical musical intervals. These results support the hypothesis that, at low pulse rates, a temporal code in the auditory nerve alone is capable of defining musical pitch.

Firing properties of spherical bushy cells in the anteroventral cochlear nucleus of the gerbil.

In gerbils, spherical bushy cells (SBCs) encode low frequency sound signals into a temporal firing pattern. To investigate the support for the timing in this temporal code, we characterized the membrane electrical properties of visually identified SBCs in brainstem slices. A brief depolarizing subthreshold transient potential (TP) triggered, with relatively invariant latency, a single spike at the onset of a response to depolarizing current pulses. The activation of a subthreshold Na+-conductance, sensitive to blockade with tetrodotoxin, and a high threshold Ca2+-conductance, sensitive to blockade with Co2+ or Cd2+, accelerated the rising phase and amplified the TP. A K+-conductance, sensitive to blockade by 4-aminopyridine (4-AP, 50 microM), shaped the decay of the TP. Following a single spike, voltage-gated activation of transient and sustained K+-conductances suppressed any tendency to repetitively discharge. A reduction in either K+-conductance due to application of 4-AP or tetraethylammonium (TEA, 10 mM), converted the single spike mode to repetitive firing during the depolarizing pulses. A persistent, tetrodotoxin-sensitive Na+-conductance amplified steady-state depolarizing responses. A hyperpolarization-activated conductance, greatly decreased by extracellular Cs+ (3 mM) but resistant to Ba2+ (up to 1 mM), filtered the responses to hyperpolarizing current inputs. A depolarized membrane potential promoted repetitive firing in SBCs. This state, expected in pathophysiological conditions, would corrupt the temporal code.

Cochlear efferent neurons projecting to both ears in the chicken, Gallus domesticus.

Different retrograde neuroanatomical tracers were injected into each cochlea of adult chicken. The number of cells labeled in the cochlear efferent cell group found bilaterally within the caudal pontine reticular formation depended upon the tracer, with True Blue and Fluoro Gold yielding maximal average counts of 332 efferent neurons per injection. Double labeling of less than 1% of these cells was possible with the combination of True Blue and Diamidino Yellow. Thus the contribution of efferent neurons with axon collaterals projecting to both ears is not fundamentally different in birds and other vertebrates.

GABA(B) receptor activation changes membrane and filter properties of auditory thalamic neurons.

Inhibitory inputs from nucleus reticularis thalami and the inferior colliculus activate gamma-aminobutyric acid B (GABA(B)) receptors in auditory thalamic neurons. These metabotropic receptors have been implicated in the oscillatory behavior of thalamic neurons. We studied the effects of the GABA(B) receptor agonist, baclofen, on membrane and filter properties of neurons in the ventral partition of the medial geniculate body (MGBv) of the rat, using whole-cell patch-clamp recording techniques in a slice preparation. Application of baclofen caused a concentration-dependent and reversible hyperpolarization of MGBv neurons. An increase in membrane conductance shunted voltage signals. The shunt suppressed firing in both tonic and burst modes which normally characterize the neuronal excitation from depolarized and hyperpolarized potentials, respectively. The GABA(B) receptor antagonist, CGP 35348 (0.5 mM), completely and reversibly blocked the baclofen-evoked hyperpolarization and increase in conductance. In voltage-clamp and during blockade of synaptic transmission with tetrodotoxin and Cd2+, baclofen activated an inwardly rectifying outward K+ current, that was sensitive to blockade with Ba2+ (0.5 mM). Intracellular applications of GTPgammaS occluded the baclofen current whereas similar applications of GDPbetaS prevented it, suggesting that G-proteins mediated the baclofen current. We measured the impedance amplitude profile in the frequency domain with swept sinusoidal current injection. MGBv neurons normally have lowpass filter characteristics at depolarized potentials and resonance at approximately 1 Hz at hyperpolarized potentials. Baclofen application reduced the impedance below 20 Hz which lowered the membrane filter quality and abolished the resonance. Despite its hyperpolarizing effect, therefore, baclofen eliminated an intrinsic tendency to oscillate as well as the intrinsic frequency selectivity of MGBv neurons.

Modulation of frequency selectivity by Na+- and K+-conductances in neurons of auditory thalamus.

In thalamic neurons, frequency-filter properties arise from intrinsic membrane properties which transform sensory inputs to thalamocortical signals. They also contribute to the tendency for the membrane to generate synchronized oscillations. We studied the frequency selectivities of thalamocortical neurons in the rat ventral medial geniculate body (MGBv) in vitro, using whole-cell recording techniques, sinewave (swept 'ZAP' or single) current inputs and pharmacological blockade of membrane currents. In a voltage range that was subthreshold to spike genesis, the frequency responses below 20 Hz were voltage-dependent, they exhibited lowpass characteristics at depolarized potentials and bandpass resonance (near 1 Hz) in the activation range (approximately -65 to -50 mV) of the low-threshold Ca2+-current (I(T)). A temperature increase of > 10 degrees C in 3 neurons did not change this voltage-dependence and increased the frequency of maximum resonance to 2 Hz. The removal of extracellular Ca2+, its equimolar substitution with Mg2+ or blockade of I(T) with Ni2+ (0.5 mM) completely blocked the resonance at hyperpolarized potentials or rest, as well as the low-threshold Ca2+-spike (LTS). Blockade of high threshold Ca2+-currents with Cd2+ (50 microM) did not affect the resonance. These data implied that, like the LTS, an activation of I(T) produced the membrane resonance. An increased ZAP-current input evoked action potentials near the resonant frequency as well as Cd2+-sensitive high-threshold Ca2+-spikes at depolarized membrane potentials and very low frequencies. By blocking a persistent Na+-current (I(NaP)), tetrodotoxin (300 nM) reduced the magnitude of the frequency response without affecting the frequency preference. The response was larger in amplitude, especially at frequencies lower than the maximum resonant frequency, when we used 4-aminopyridine (0.05-0.1 and 1-2 mM), Ba2+ (0.2 mM) or Cs+ (3 mM) to block voltage-dependent K+-currents. From these data, we suggest that A-type (I(A) and I(As)) and inwardly rectifying (I(KIR)) K+-currents modulate resonance, changing the quality of the lowpass filter function. We conclude that the generation of membrane resonance in MGBv neurons depends critically on I(T)-activation while the quality of the frequency response is subject to modulation by voltage-dependent conductances. The frequency selectivities in MGBv may contribute to lowpass filter functions for auditory transmission during wakefulness and oscillations observed during sleep.

Retrograde transport of [3H]-GABA by lateral olivocochlear neurons in the rat.

Injection of [3H]-gamma aminobutyric acid (GABA) into the perilymphatic space of the rat's inner ear resulted in retrograde labeling of a portion of the small efferent olivocochlear neurons within the lateral superior olivary nucleus (LSO). These cells were of similar size as LSO neurons stained immunohistochemically with antibodies to the GABA synthesizing enzyme glutamic acid decarboxylase (GAD). They were of fusiform shape, but smaller than principal LSO cells, which did not stain for GAD and did not accumulate [3H]-GABA. Other efferent cochlear and vestibular neurons were not labeled.

Location of motoneurons innervating the middle ear muscle of the chicken, (Gallus domesticus).

The motoneuron pool for the musculus columellae, the avian equivalent to the m. stapedius, was identified by retrograde labeling with WGA-HRP. It consists of a discrete group of approximately 65 neurons located along the dorsolateral border in the ventral subnucleus of the facial nuclear complex. Other facial motoneurons were only labeled when diffusion of the tracer into neighbor structures was not excluded. The dorsal subnucleus of the facial nerve innervates the m. depressor mandibulae.

Phrenic nerve reinnervation of the cat's larynx: a new technique with proven success.

Reinnervation of the posterior cricoarytenoid muscle (PCA) should provide vocal cord abduction on inspiration, and passive adduction to enable phonation. Previous investigators have shown that reinnervation is possible, but results have not been clinically encouraging. When reinnervation was successful, the question remained whether it was provided by the transplanted nerve or by the ingrowth of adjacent nerves. In this study the phrenic nerve was transplanted directly into the PCA in a series of 12 cats. Fibrin glue was used to overcome nerve trauma and to prevent retraction of the nerve from the PCA. Laryngoscopy, electromyography, and retrograde labeling of the phrenic motoneurons provided evidence of functional reinnervation in 9 cats. Partial or complete failure in the remaining 3 was due to retraction of the nerve from the muscle. These results appear to justify trials of the procedure in humans.

Projection of afferents from individual vestibular sense organs to the vestibular nuclei in the pigeon.

The projection of individual labyrinthine sensory organs to the brain stem was studied by autoradiography, employing discrete [3H]leucine injections into the sensory epithelia. Within the vestibular nuclei, separate partly overlapping termination areas for each end organ were found in the superior and descending vestibular nuclei, whereas projection territories in the medial, ventrolateral and tangential nuclei overlapped extensively. A few lagenar fibres terminated in the external cuneate nucleus. Semicircular canals and utricular macula also project to the lateral cerebellar nucleus and the reticular formation. For each semicircular canal a projection system could be traced to distinct subgroups of the extraocular motoneuron pools.

Bechterew decompensation.

A right labyrinthectomy was performed in rats 5 months after a left labyrinthectomy. Spontaneous compensation of balance functions after both operations was assessed by observing nystagmus, rolling about the longitudinal axis, circular walking and head tilt. Decompensation, induced by brief halothane-NO anaesthesia, released predominantly symptoms characteristic for the period after the first labyrinthectomy. Bechterew symptoms could, however, also be decompensated. It is concluded that Bechterew compensation does not re-establish balance in the central vestibular system. Dysbalance and vertigo of vestibular origin is thus conceivable in patients after all peripheral vestibular function has been lost.

Vestibular evoked potentials in thalamus and basal ganglia of the squirrel monkey (Saimiri sciureus).

In anesthetized squirrel monkeys vestibular representation in the thalamus and basal ganglia was determined by field potential recording using peripheral electrical vestibular nerve stimulation. Vestibular thalamic regions were investigated for cortical connections. Two relatively large thalamic areas, nucleus ventralis posterolateralis, VPL and the posterior nuclear group (Po) received vestibular inputs with short latencies suggesting direct connections with the vestibular nuclei. Antidromic stimulation of the area 3 a vestibular field did not produce responses in any of the vestibular thalamic fields. The vestibular regions in VPL and Po can be antidromically invaded from SI and the anterior parietal lobe respectively. In the striatum vestibular fields were found in the suprathalamic portion of the nucleus caudatus and dorsomedially in the putamen.

Metabotropic transmitter actions in auditory thalamus.

Neurons in the ventral partition of the medial geniculate body (MGBv), the primary auditory thalamus, receive afferent input from the inferior colliculus via excitatory glutamate-ergic and inhibitory GABA-ergic input fibres. The feedback from the auditory cortex to the thalamic relay also is mediated via neuron systems using glutamate and GABA as transmitters. We studied effects on excitability mediated by these transmitters via G-protein coupled metabotropic receptors. In a slice preparation of rat thalamus we investigated the membrane responses of MGBv neurons using the whole cell recording technique. Application of a metabotropic glutamate receptor (mGluR) agonist, ACPD (5-100 microM), depolarized MGBv neurons. As a result, the burst mode of firing, which characterizes states of sleep at hyperpolarized potentials was replaced by the tonic mode, which is compatible with sound signal transmission during alertness. The depolarization was caused by an inward current (I(ACPD)) that persisted during blockade of Na+ channels with tetrodotoxin (TTX) and of Ca2+ channels with Cd2+. The I(ACPD) depended, however, on extracellular Na+, which could be replaced with Li+, excluding a major contribution of the Na+/Ca2+ exchange current. ACPD application also inhibited an inwardly rectifying K+ current at hyperpolarized potentials and activated an outward current in the depolarized range. Application of the GABA(B) agonist, baclofen (10 microM), hyperpolarized MGBv neurons by activation of an inwardly rectifying K+ current. The corresponding membrane conductance acted as a powerful shunt that reduced voltage responses and inhibited firing in both the tonic and burst modes. Thus, the effects of GABA(B) receptor activation would suppress auditory signal transfer, whereas mGluR activation enhances excitability, possibly accounting for the alerting effects of certain auditory stimuli.

Firing modes and membrane properties in lemniscal auditory thalamus.

In neurons of the auditory thalamus, patterned sequences of action potentials encode the features of sound stimuli. The patterns vary with the membrane potential, characterizing states of wakefulness and sleep. We studied the dependence of the patterns on the membrane potential and specific voltage-gated conductances, using whole-cell patch-clamp recordings from neurons in the ventral medial geniculate body (MGBv) of in vitro slices. Thalamocortical neurons, identified with neurobiotin, exhibited different firing patterns to an excitatory input, depending on the initial membrane potential. From depolarized potentials, the neurons fired in a tonic mode. The delay to firing in this mode was regulated by a balance of persistent Na+ and A-type K+ conductances. When transiently depolarized from hyperpolarized holding potentials, the neurons fired brief phasic responses (burst mode). Phasic responses were induced by low threshold Ca2+ spikes (LTSs); the LTS-amplitude was controlled by Na+ and K+ conductances. Under favourable conditions, an LTS triggered more than one action potential and one or more high threshold Ca2+ spikes (HTSs). Consciously perceived sound signals are transmitted in the tonic mode. During sleep, alerting stimuli may interact with membrane non-linearities, converting hyperpolarized bursting MGBv neurons to the tonic mode.