Correspondences between afferent innervation patterns and response dynamics in the bullfrog utricle and lagena

Otoconial afferents in the bullfrog were characterized as gravity or vibratory sensitive by their resting activity and their responses to head tilt and vibration. The responses of gravity afferents to head tilt were tonic, phasic-tonic, or phasic. A few afferents, termed vibratory/gravity afferents, had gravity as well as vibratory sensitivity. Functionally identified otoconial afferents were injected with Lucifer Yellow and subsequently traced to their peripheral arborizations. Morphological maps, previously constructed with the scanning electron microscope, were used to identify microstructural features of the sensory maculae associated with the peripheral arborizations of dye-filled afferents. The utricular and lagenar macula each is composed of a specialized central band surrounded by a peripheral field. The central bands are composed of densely packed medial rows and more sparsely packed lateral rows of hair cells. Hair cells exhibit a variety of surface topographies which correspond with their macular location. The response dynamics of afferents in the utricle and lagena correspond with the macular locations of their peripheral arborizations. Tonic afferents were traced to hair cells in the peripheral field. Phasic-tonic and phasic afferents innervated hair cells in the lateral rows of the central band, the former innervating hair cells at the edges of the central band and the latter innervating hair cells located more medially. Afferents with vibratory sensitivity were traced to hair cells in the medial rows of the lagenar central band. The response dynamics of afferents corresponded with the surface topography of their innervated hair cells. Tonic and phasic-tonic gravity afferents innervated hair cells with stereociliary arrays markedly shorter than their kinocilium (Lewis and Li types B and C) while phasic gravity and vibratory afferents innervated hair cells with stereociliary arrays nearly equal to their kinocilium (Lewis and Li types E and F). Vibratory sensitivity was uniquely associated with hair cells possessing bulbed kinocilium (Lewis and Li type E) while afferents sensitive to both gravity and vibration innervated hair cells from both of the above groups. We argue that afferent response dynamics are determined, at least in part, at the level of the sensory hair bundle and that morphological variations of the kinocilium and the otoconial membrane are dictated by specialization of sensitivity. We propose that morphological variations of the kinocilium reflect variations in its viscoelastic properties and that these properties determine the nature of the mechanical couple between the stereociliary array and the otoconial membrane.     

Phasic and tonic firing properties in rat oculomotor nucleus motoneurons, studied in vitro

Alert-chronic studies show that ocular motoneurons (Mns) exhibit a phasic and tonic firing correlated with eye saccade-velocity and position (fixation), respectively. Differences in the phasic and tonic firing among Mns depend on synaptic inputs and/or the intrinsic membrane properties. We have used in vitro slice preparation to investigate the contribution of membrane properties to firing properties of Wistar rat oculomotor nucleus Mns. We recorded different discharge patterns and focused on Mns with sustained discharge (type I) because they were the most abundant, and their firing pattern resembles that reported in alert preparations. Various differences divided these Mns into types I(A) and I(B); the afterhyperpolarization (AHP) phase of the spike was monophasic in I(A) and biphasic in I(B); I(A) Mns showed tonic or phasic-tonic firing depending on the current intensity, while I(B) Mns showed phasic-tonic discharge; the phasic firing was higher in I(B) than in I(A) Mns; I(A) Mns fired in a narrower range than did I(B) Mns; and I(A) Mns showed lower maximum frequency than did I(B) Mns. In conclusion, I(A) and I(B) Mns show different phasic firing properties and dynamic range, supported by intrinsic membrane properties. We suggest that I(A) and I(B) Mns innervate fast-twitch muscle fibres with different contraction speeds, and could contribute to generating a fine phasic signal for a graded muscle contraction. Finally, we have demonstrated an inverse relationship between Mn thresholds and tonic firing gain, concluding that intrinsic membrane properties could not support the covariation between tonic firing gain and recruitment thresholds reported in alert studies.     

Laryngeal afferent inputs during vocalization in the cat.

To reveal the kind of information about the larynx which is transmitted to the central nervous system during vocalization, we studied discharge patterns of single fibers of the laryngeal afferent nerve during electrically induced vocalization in ketamine-anesthetized cats. Recorded fibers were classified into four types based on their discharge patterns. Type A fibers responded to vocal fold vibration during vocalization. Type B fibers increased their activity during vocalization without synchronization with vocal fold vibration. Type C fibers decreased their activity during vocalization. Type D fibers discharged only at the onset of vocal fold adduction and abduction. We discuss the functional properties of these afferents and the possibility that these afferent inputs participate in the feedback control of vocalization.     

Peripheral innervation patterns of vestibular nerve afferents in the bullfrog utriculus

Vestibular nerve afferents innervating the bullfrog utriculus differ in their response dynamics and sensitivity to natural stimulation. They also supply hair cells that differ markedly in hair bundle morphology. To examine the peripheral innervation patterns of individual utricular afferents more closely, afferent fibers were labeled by the extracellular injection of horseradish peroxidase (HRP) into the vestibular nerve after sectioning the vestibular nerve medial to Scarpa's ganglion to allow the degeneration of sympathetic and efferent fibers. The peripheral arborizations of individual afferents were then correlated with the diameters of their parent axons, the regions of the macula they innervate, and the number and type of hair cells they supply. The utriculus is divided by the striola, a narrow zone of distinctive morphology, into medial and lateral parts. Utricular afferents were classified as striolar or extrastriolar according to the epithelial entrance of their parent axons and the location of their terminal fields. In general, striolar afferents had thicker parent axons, fewer subepithelial bifurcations, larger terminal fields, and more synaptic endings than afferents in extrastriolar regions. Afferents in a juxtastriolar zone, immediately adjacent to the medial striola, had innervation patterns transitional between those in the striola and more peripheral parts of the medial extrastriola. Most afferents innervated only a single macular zone. The terminal fields of striolar afferents, with the notable exception of a few afferents with thin parent axons, were generally confined to one side of the striola. Hair cells in the bullfrog utriculus have previously been classified into four types based on hair bundle morphology (Lewis and Li: Brain Res. 83:35–50, 1975). Afferents in the extrastriolar and juxtastriolar zones largely or exclusively innervated Type B hair cells, the predominant hair cell type in the utricular macula. Striolar afferents supplied a mixture of four hair cell types, but largely contacted Type B and Type C hair cells, particularly on the outer rows of the medial striola. Afferents supplying more central striolar regions innervated fewer Type B and larger numbers of Type E and Type F hair cells. Striolar afferents with thin parent axons largely supplied Type E hair cells with bulbed kinocilia in the innermost striolar rows. © 1994 Wiley-Liss, Inc.     

Central projections of the utricular nerve in the gerbil

The central projections of primary afferent fibers in the utricular nerve, which convey linear head acceleration signals to neurons in the brainstem and cerebellum, are not completely defined. The purpose of this investigation was twofold: 1) to define the central projections of the gerbil utricular afferents by injecting horseradish peroxidase (HRP) and biotinylated dextran amine (BDA) into the utricular macula; and 2) to investigate the projections of individual utricular afferents by injecting HRP intracellularly into functionally identified utricular neurons. We found that utricular afferents in the gerbil projected to all divisions of the vestibular nuclear complex, except the dorsal lateral vestibular nucleus. In addition, terminals were observed in the interstitial nucleus of the eighth nerve, nucleus Y, external cuneate nucleus, and lobules I, IV, V, IX, and X of the cerebellar vermis. No projections appeared in the flocculus or paraflocculus. Fibers traversed the medial and intermediate cerebellar nuclei, but terminals appeared only occasionally. Individual utricular afferents collateralize extensively, projecting to much of the brainstem area innervated by the whole of the utricular nerve. This study did not produce complete filling of individual afferent collateral projections into the cerebellar cortex.     

Inhibition of nociceptive responses of wide-dynamic-range neurons by peripheral nerve stimulation

Of 107 neurons from the sacral and coccygeal levels of the spinal cord in anesthetized intact rats examined, 62 wide-dynamic-range (WDR) neurons that responded to noxious heating of the tail were recorded. On the basis of their inhibitory responses through A-beta or A-delta afferent fibers to noxious stimulation, these neurons were classified into one of the following three types: Type I--neurons inhibited only by A-beta afferent nerve impulses; Type II--neurons inhibited only by A-delta afferent nerve impulses; Type III--neurons inhibited by both. The present results are compared with previously reported behavioral results.     

Tonal response patterns of primary auditory cortex neurons in alert cats

The firing rates of primary auditory cortex (A1) neurons are known to be modulated only at the onset, offset, and change of a tonal stimulus in anesthetized animals. The tonal response pattern has been rarely investigated in alert animals. We investigated the time-course of A1 neuron responses to a steady tonal stimulus in alert cats. We found four types of firing responses based on statistical evaluation of the time course of the firing rate. The tonic cells (38 cells) showed a significant (P<0.05) firing increase throughout the stimulus period after a relatively long latency (mean, 25.3 ms) with little tendency of adaptation. The phasic-tonic cells (22 cells) showed a significant firing increase throughout the stimulus period after a medium latency (19.8 ms) with tendency of adaptation to less than a half of the maximum excitation level. Phasic cells (15 cells) responded, after a short latency (10.2 ms), at onset and offset of the stimuli. The unresponsive cells (26 cells) did not show a significant firing increase during stimuli. The findings suggest that there is a functional difference between each type of cells: the tonic cells encode information of static auditory signals in their firing rates; the phasic-tonic cells, of the changing auditory signal during the stimulus period; and the phasic cells, of rapid change of the auditory signal at onset and offset.     

Peripheral patterns of terminal innervation of vestibular primary afferent neurons projecting to the vestibulocerebellum in the gerbil

Retrograde transganglionic labeling techniques with biotinylated dextran amine (BDA) were used to examine the terminal field structure and topographical patterns of innervation within the vestibular sensory end organs of vestibular primary afferent neurons projecting to the cerebellar uvula/nodulus and flocculus lobules in the gerbil. Robust, dark labeling in the cristae ampullares suggested that the vast majority of the terminals of afferent neurons were of the dimorphic type. The majority (94% to the uvula/nodulus and 100% to the flocculus) innervates the peripheral zones of each of the three semicircular canal cristae. Comparison of the type and distribution of terminals across the canalicular sensory neuroepithelium with morphophysiological studies in chinchilla suggests that the labeled population consists predominantly of peripheral terminal fields of lower-to-intermediate gain, more regularly firing, tonic afferents. For otolith organ-related afferents, the uvula/nodulus receives strong inputs from primary otolith afferent neurons identified as dimorphic in type that predominately innervate the peristriolar zones of the utricular and saccular maculae. No direct otolith organ-related inputs to the flocculus were observed. In contrast to the canal afferents, the types and locations of labeled otolith afferent terminals suggest that they largely consist of irregularly firing, high-gain, phasic neurons.     

Phase-modulated antidromic action potentials in the dorsal root of the rat during locomotion

Discharges of antidromic spike potentials (ASP) were recorded from the rat dorsal root during locomotion. Tonic and phasic components were revealed in discharges of ASP. The phase-dependent modulation of phasic component was observed during locomotion. The role of ASP as a possible mechanism for restriction of afferent input was discussed.     

Spike encoding of neurotransmitter release timing by spiral ganglion neurons of the cochlea

Mammalian cochlear spiral ganglion neurons (SGNs) encode sound with microsecond precision. Spike triggering relies upon input from a single ribbon-type active zone of a presynaptic inner hair cell (IHC). Using patch-clamp recordings of rat SGN postsynaptic boutons innervating the modiolar face of IHCs from the cochlear apex, at room temperature, we studied how spike generation contributes to spike timing relative to synaptic input. SGNs were phasic, firing a single short-latency spike for sustained currents of sufficient onset slope. Almost every EPSP elicited a spike, but latency (300-1500 μs) varied with EPSP size and kinetics. When current-clamp stimuli approximated the mean physiological EPSC (≈300 pA), several times larger than threshold current (rheobase, ≈50 pA), spikes were triggered rapidly (latency, ≈500 μs) and precisely (SD, <50 μs). This demonstrated the significance of strong synaptic input. However, increasing EPSC size beyond the physiological mean resulted in less-potent reduction of latency and jitter. Differences in EPSC charge and SGN baseline potential influenced spike timing less as EPSC onset slope and peak amplitude increased. Moreover, the effect of baseline potential on relative threshold was small due to compensatory shift of absolute threshold potential. Experimental first-spike latencies in response to a broad range of stimuli were predicted by a two-compartment exponential integrate-and-fire model, with latency prediction error of <100 μs. In conclusion, the close anatomical coupling between a strong synapse and spike generator along with the phasic firing property lock SGN spikes to IHC exocytosis timing to generate the auditory temporal code with high fidelity.     

Synaptic transmission between single tactile and kinaesthetic sensory nerve fibers and their central target neurones

Selective activation of single tactile or kinaesthetic afferent nerve fibers in conscious human subjects by means of the intraneural microstimulation procedure reveals quite marked differences among the different classes in their capacity for eliciting perceptual responses. This work, conducted largely by Swedish researchers, suggests that there may be differential transmission security for different fiber classes across synaptic linkages in the central tactile and kinaesthetic sensory pathways. In order to test this hypothesis we have developed an experimental paradigm in the anaesthetized cat, based upon paired, simultaneous recording from an individual afferent fiber in an intact peripheral nerve fascicle, and from the central target neurone of that afferent fiber within the dorsal column nuclei (DCN). Our results demonstrate, for all tactile and kinaesthetic fiber classes examined, that the minimum sensory input, a single impulse in one sensory fiber, can generate spike output from DCN target neurones. This remarkable security of transmission has been demonstrated for single Pacinian corpuscle (PC) fibers (associated with Pacinian Corpuscle receptors); single SAI and SAII fibers (the slowly adapting Type I and II tactile fibers associated, respectively, with Merkel and Ruffini receptor endings); the Hair Follicle Afferent fibers (HFA fibers); and kinaesthetic afferent fibers of both joint and muscle origin. The results demonstrate that the differential capacities of various tactile and kinaesthetic fiber classes to generate perceptual responses when activated singly in microneurography experiments do not appear to be explicable in terms of systematic differences in DCN transmission characteristics.     

Irregular primary otolith afferents from the guinea pig utricular and saccular maculae respond to both bone conducted vibration and to air conducted sound

This study sought to identify in guinea pig the peripheral sense organ of origin of otolith irregular primary vestibular afferent neurons having a very sensitive response to both air-conducted sound (ACS) and bone-conducted vibration (BCV). Neurons responding to both types of stimuli were labelled by juxtacellular labelling by neurobiotin. Whole mounts of the maculae showed that some vestibular afferents activated by both ACS and BCV originate from the utricular macula and some from the saccular macula - there is no "afferent specificity" by one sense organ for ACS and the other for BCV - instead some afferents from both sense organs have sensitive responses to both stimuli. The clinical implication of this result is that differential evaluation of the functional status of the utricular and saccular maculae cannot rely on stimulus type (ACS vs BCV), however the differential motor projections of the utricular and saccular maculae allow for differential evaluation of each sense organ.     

Firing properties of frog tectal neurons in vitro

Whole-cell recordings from frog tectal slices revealed different types of neuronal firing patterns in response to prolonged current injection. The patterns included regular spiking without adaptation, accelerating firing, adapting spiking, repetitive bursting and phasic response with only one spike. The observed firing patterns are similar to those found in the mammalian superior colliculus. The frog tectum could be a useful preparation in elucidating the relationship between neuronal function and membrane properties.     

Posture-correlated responses to vestibular polarization in vermal versus intermediate Posterior cerebellar cortex

Cerebellar lesion experiments have led to the concept that the medial longitudinal zone controls postural tone while the intermediate zone controls discrete movement. This study is a test of the hypothesis that, of the two zones, the medial zone is more closely linked to the resting discharge of vestibular afferent fibers, a prime source of neural tonus underlying the tonus of posture. Unilateral polarizations of the vestibular apparatus via the round window in awake, unrestrained guinea pigs caused step changes of postural attitude, the direction of which was polarity dependent. In anesthetized animals, these currents caused nonadapting step changes, or posture-correlated responses in the level of resting discharge in vestibular primary afferent fibers. In the medial and the intermediate cerebellar cortices of the posterior lobe, the proportion of step-like responses was similar, in contradiction to the hypothesis. This suggests that the cerebellar computations for controlling both postural tonus and discrete movements require information about vestibular tonus in terms of simple spike activity.     

Intracellular study of rat entopeduncular nucleus neurons in an in vitro slice preparation: electrical membrane properties

Electrical properties of rat entopeduncular nucleus (EP) neurons were studied in vitro using slice preparations. Of 108 EP neurons recorded, 104 were classified into two types based on their membrane properties. Type I neurons (n = 86) possessed: (1) a strong, time-dependent anomalous rectification that was sensitive to Cs⁺; (2) a weak spike adaptation; and (3) a strong rebound excitation with a low threshold Ca-spike and fast spikes. Many Type I neurons displayed spontaneous repetitive firing. Some of them generated spontaneous Ca-dependent plateau potentials with fast spikes upon application of tetraethylammonium bromide. Type II neurons (n = 18) had: (1) no apparent rectification; (2) a strong spike adaptation; and (3) a ramp-shaped repolarization, similar to the A-current, at the offset of a hyperpolarizing pulse. Features common to both types included: (1) a similar range of the input resistance; (2) capability of generating high threshold Ca-spike; and (3) generation of postactive hyperpolarizations (i.e. Ca-activated K-conductance). The great majority (Type I) of rat EP neurons share similar electrical properties. A minority of neurons (Type II) behave differently from Type I neurons and share similar properties among themselves.     

Spatial and morphological differentiation of trigger zones in afferent fibers to the teleost utricle.

A morphological correlate of the trigger site (the locus of action potential initiation) was identified in afferent axons of the utricle in the ear of two species of teleost fish. These sites were identified by the ferric-ferrocyanide (Prussian blue) cytochemical procedure and they were correlated with the geometries of afferent intraepithelial arbors as visualized by means of a silver stain. The intraepithelial arbors of afferent fibers show regional distributions that correlate with axon diameter and Prussian blue staining. Afferent axons with diameters greater than 4-5 microns only innervate the striola regions of the epithelium and terminate as one of two distinct types of intraepithelial arbors. Afferent axons with diameters smaller than 4 microns are ubiquitously distributed throughout the epithelium. Arbors that stained by Prussian blue within the utricular epithelium are restricted to the striolar regions. These arbors possess nodal-like membrane in different branches as postsynaptic membrane. Afferents that innervate hair cells in the extrastriolar epithelial regions stained with Prussian blue only at the extraepithelial terminal heminode. The postsynaptic membrane of these afferents is passive or dendritic-like.     

Thermosensitive and osmoreceptive afferent fibers in the hepatic branch of the vagus nerve

The hepatic vagus nerve contains various thermosensitive afferent fibers which are widely varied in their sensitivity. Their Q10 values lie between 4 and 16. The discharge rate is positively correlated with increase of liver temperature (warm fiber type). The result supports the existence of a thermosensitive structure in the liver which may possibly contribute to maintain thermal homeostasis. Neural responses to the osmotic changes in the perfusion solution have been analyzed. It was found that two different types of osmosensitive afferent fibers exist in the hepatic vagus; one is characterized by increasing the frequency of spike discharges in response to higher osmotic pressure, while the other shows the same response to lowered levels. Behavioral changes caused by hepatic vagotomy were observed. These results provide evidence for the existence of an osmoreceptor mechanism. The role of these hepatic afferent nerves in homeostasis are briefly discussed.     

Role of metabosensitive afferent fibers in neuromuscular adaptive mechanisms

Role of metabosensitive afferent fibers in neuromuscular adaptive mechanisms. Adaptation to exercise is provided by central neuron activity adjustments which are regulated partly by activation of group I and II (mechanosensitive) and group III and IV (metabosentitive) afferent fibers. These last two groups are activated by exercise-induced changes in muscle metabolism. The role played by these afferents seems to be crucial to exercise and fatigue tolerance adaptive mechanisms. Nevertheless, many questions remain unresolved. The aim of this review is to focus on the involvement of metabosensitivity in sensorimotor loops and neuromuscular adaptive mechanisms. The existence of an adaptive cardiovascular and respiratory reflex to exercise originating from metabosensitive afferent fiber activation is well established. Furthermore, the mechanism of skeletal muscle protection against fatigue could be due to modulation of central motor command at the spinal and supraspinal levels via these afferent fibers.     

Intrinsic membrane properties and morphological characteristics of interneurons in the rat supratrigeminal region

The membrane properties and morphological features of interneurons in the supratrigeminal area (SupV) were studied in rat brain slices using whole-cell patch clamp recording techniques. We classified three morphological types of neurons as fusiform, pyramidal, and multipolar and four physiological types of neurons according to their discharge pattern in response to a 1-sec depolarizing current pulse from -80 mV. Single-spike neurons responded with a single spike, phasic neurons showed an initial burst of spikes and were silent during the remainder of the stimulus, delayed-firing (DF) neurons exhibited a slow depolarization and delay to initial spike onset, and tonic (T) neurons showed maintained a discharge throughout the stimulus pulse. In a subpopulation of neurons (10%), membrane depolarization to around -44 mV produced a rhythmic burst discharge (RB) that was associated with voltage-dependent subthreshold membrane oscillations. Both these phenomena were blocked by the sodium channel blocker riluzole at a concentration that did not affect the fast transient spike. Low doses of 4-AP, which blocks low-threshold K+ currents, transformed bursting into low-frequency tonic discharge. In contrast, bursting occurred with exposure to cadium, a calcium-channel blocker. This suggests that persistent sodium currents and low-threshold K+ currents have a role in intrinsic burst generation. Importantly, RB cells were most often associated with multipolar neurons that exhibited either a DF or a T discharge. Thus, the SupV contains a variety of physiological cell types with unique morphologies and discharge characteristics. Intrinsic bursting neurons form a unique group in this region. .     

5‐HT3A‐driven green fluorescent protein delineates gustatory fibers innervating sour‐responsive taste cells: A labeled line for sour taste?

Taste buds contain multiple cell types with each type expressing receptors and transduction components for a subset of taste qualities. The sour sensing cells, Type III cells, release serotonin (5‐HT) in response to the presence of sour (acidic) tastants and this released 5‐HT activates 5‐HT₃ receptors on the gustatory nerves. We show here, using 5‐HT_3AGFP mice, that 5‐HT₃‐expressing nerve fibers preferentially contact and receive synaptic contact from Type III taste cells. Further, these 5‐HT₃‐expressing nerve fibers terminate in a restricted central‐lateral portion of the nucleus of the solitary tract (nTS)—the same area that shows increased c‐Fos expression upon presentation of a sour tastant (30 mM citric acid). This acid stimulation also evokes c‐Fos in the laterally adjacent mediodorsal spinal trigeminal nucleus (DMSp5), but this trigeminal activation is not associated with the presence of 5‐HT₃‐expressing nerve fibers as it is in the nTS. Rather, the neuronal activation in the trigeminal complex likely is attributable to direct depolarization of acid‐sensitive trigeminal nerve fibers, for example, polymodal nociceptors, rather than through taste buds. Taken together, these findings suggest that transmission of sour taste information involves communication between Type III taste cells and 5‐HT₃‐expressing afferent nerve fibers that project to a restricted portion of the nTS consistent with a crude mapping of taste quality information in the primary gustatory nucleus.