Synaptic events and discharge patterns of cochlear nucleus cells. II. Frequency-modulated tones.

Responses of 99 cochlear nucleus cells and 24 cochlear nerve fibers were studied with FM signals; 14 cochlear nerve fibers and 57 cochlear nucleus cells were studied at four rates of modulation and several signal intensities. Classification of FM response patterns as symmetrical, asymmetrical, or unidirectional was based on the calculation of a symmetry factor (S), which compared the number of discharges evoked by the ascending and by the descending phases of the FM sweep. Certain FM response patterns could not adequately be described by the symmetry factor along and variables of modulation rate and signal intensity had significant influence. A correspondence was found between the four response classes evoked by a steady-frequency tone burst (primarylike, buildup, onset, and pause) and the FM response pattern. Cochlear nerve fibers showed symmetrical response patterns to FM stimulation. Primarylike units were similar to eighth nerve fibers and generally showed symmetrical FM responses. Occasional eighth nerve fibers and primarylike cells developed asymmetry at the fastest rate of modulation (50 sps). Buildup units showed a variety of response patterns to FM signals. Onset units generally showed asymmetrical response patterns with the greater response occurring to the ascending than to the descending phase of the FM sweep. Pause units showed a characteristic inhibition of activity at 5 sps (rate-dependent inhibition). Of the 57 cochlear nuclear cells studied in response to FM signals, 16 were symmetrical, another 16 were symmetrical except at the fastest modulation rate, 12 were asymmetrical, 3 were unidirectional, and 10 showed complex responses to certain signal rates or intensities. It is clear the the cat cochlear with its complex cytoarchitecture is involved in the recoding of acoustic information. Some units in cochlear nucleus demonstrate differential responses to the direction and to the rate of frequency movement. Other cochlear nucleus cells respond as eighth nerve fibers and may serve as simple "relays" in transmitting information from the cochlea to higher auditory centers.     

Statistical analysis and interpretation of the initial response of cochlear nucleus neurons to tone bursts

1. Subject of investigation is the initial response of cochlear nucleus neurons and units presumed to be auditory nerve fibres to CF tone burst stimulation. 2. The initial response is characterized by computing the distribution of the latency of the first spike and of the duration of the first interval in the ensemble of responses to a large number of stimuli. 3. In many of the neurons the properties of both distributions appear to be related. The presumed auditory nerve fibres and spontaneously active cochlear nucleus neurons showing only activation responses to tonal stimuli (A type) exhibit irregularity in both response onset and intervals. Minimum latency and minimum first intervals are short. On the other hand, spontaneously active neurons with both activation and suppression in the response area (AS type) and silent neurons showing only activation (A(S) type) often show a more precisely timed onset of response and narrow interval distributions. In many neurons this leads to oscillations in the PSTH (chopping). In these neurons minimum latency and minimum first interval have higher values. The longer minimum latency cannot be attribute-d to longer pure time delays in these neurons. 4. The results are interpreted as speaking in favour of temporal integration as an important mechanism in many of the AS and A(S) neurons, particularly those in the DCN. The firing patterns of A neurons are thought to indicate virtual absence of this mechanism. 5. Using pure time delay estimates derived from cross-correlation functions, computed from the responses to stationary noise, an attempt is made to estimate the integration time in the cochlear and in the cochlear nucleus neurons.     

Synaptic influences of pontine nuclei on cochlear nucleus cells

Using the in vitro isolated whole brain preparation of the guinea pig, we tested the synaptic effects induced by the stimulation of pontine nuclei (PN) in intracellularly recorded and stained principal cells of the cochlear nucleus (CN). Twenty percent of the recorded cells in all CN subdivisions responded to stimulation of either ipsilateral or contralateral PN, and 12% of the cells exhibited convergence of inputs from both sides. The responses were recorded only in stellate cells of the ventral CN and in the pyramidal cells of the dorsal CN, whereas no responses were observed in bushy, octopus, and giant cells. PN stimulation produced excitatory and inhibitory postsynaptic potentials as well as mixed responses. The heterogeneous nature and the wide latency range (3.2–18 ms) of observed responses suggest significant variability in the underlying synaptic mechanisms and the implicated pathways. We propose that PN projections to the CN, terminating mainly in the granule cell domain (GCD), together with other non-auditory and auditory inputs contribute to multimodal convergence in the GCD leading ultimately to modulatory actions on the output activity of CN principal cells.     

Synaptic inputs to granule cells of the dorsal cochlear nucleus

The mammalian dorsal cochlear nucleus (DCN) integrates auditory nerve input with nonauditory signals via a cerebellar-like granule cell circuit. Although granule cells carry nonauditory information to the DCN, almost nothing is known about their physiology. Here we describe electrophysiological features of synaptic inputs to granule cells in the DCN by in vitro patch-clamp recordings from P12 to P22 rats. Granule cells ranged from 6 to 8 microm in cell body diameter and had high-input resistance. Excitatory postsynaptic currents consisted of both AMPA receptor-mediated and N-methyl-D-aspartate receptor-mediated currents. Synaptically evoked excitatory postsynaptic currents ranged from -25 to -140 pA with fast decay time constants. Synaptic stimulation evoked both short- and long-latency synaptic responses that summated to spike threshold, indicating the presence of a polysynaptic excitatory pathway in the granule cell circuit. Synaptically evoked inhibitory postsynaptic currents in Cl(-)-loaded cells ranged from -30 to -1,021 pA and were mediated by glycine and, to a lesser extent, GABA(A) receptors. Unlike cerebellar granule cells, DCN granule cells lacked tonic inhibition by GABA. The glycinergic synaptic conductance was mediated by heteromeric glycine receptors and was far stronger than the glutamatergic conductance, suggesting that glycinergic neurons may act to gate nonauditory signals in the DCN.     

Sensitivities of cells in anteroventral cochlear nucleus of cat to spatiotemporal discharge patterns across primary afferents

1. This study tested the hypothesis that a cell in the anteroventral cochlear nucleus (AVCN) that receives convergent input from auditory nerve (AN) fibers can be sensitive to the temporal pattern of discharges on the set of AN fibers providing its input. 2. The temporal discharge pattern across the population of low-frequency AN fibers was manipulated by varying the phase spectra of complex stimuli that had fixed, flat magnitude spectra. By introducing a phase shift with variable slope at a particular frequency, the relative times of discharge of phase-locked neurons with different characteristic frequencies (CFs) could be varied. In this manner the overall spatiotemporal discharge pattern across the array of AN fibers was systematically manipulated. 3. Some low-frequency cells in the AVCN were sensitive to changes in the slope of the phase transition of the complex stimulus. The cells that were sensitive came from several different cell types in the AVCN. Their responses were consistent with the hypothesis that these cells were sensitive to the temporal relationships between discharges on their primary inputs and that they received inputs with different CFs, because the phase shifts introduced relative time differences between different frequencies. 4. Other cells were not sensitive to the degree of phase shift of the stimulus. This insensitivity implied either that these cells received inputs of the same, or nearly the same, CF, or that they were not sensitive to the time differences introduced by these changes in the phase spectra, or both. 5. The cells that were sensitive to the manipulations of the phase spectrum were located in the posterior region of anterior AVCN and in the posterior region of AVCN and thus were presumably either globular bushy, small spherical bushy, or stellate cells. No sensitive cells were located in the most anterior region of the AVCN, where large spherical bushy cells are located. 6. Temporal discharge patterns across the AN population in response to complex stimuli change as a function of level. Accordingly, the sensitivity of neurons to changes in the phase transitions of the complex stimuli used in this study was often affected by the level of the stimulus. 7. The sensitivity to changes in the phase spectrum was a frequency-specific effect. That is, a cell was most sensitive to changes made in phase that were centered near its CF and less sensitive to changes in phase that were introduced at frequencies below or above CF.(ABSTRACT TRUNCATED AT 400 WORDS)     

Synaptic physiology in the cochlear nucleus angularis of the chick

Nucleus angularis (NA), one of the two cochlear nuclei in birds, is important for processing sound intensity for localization and most likely has role in sound recognition and other auditory tasks. Because the synaptic properties of auditory nerve inputs to the cochlear nuclei are fundamental to the transformation of auditory information, we studied the properties of these synapses onto NA neurons using whole cell patch-clamp recordings from auditory brain stem slices from embryonic chickens (E16-E20). We measured spontaneous excitatory postsynaptic currents (EPSCs), and evoked EPSCs and excitatory postsynaptic potentials (EPSPs) by using extracellular stimulation of the auditory nerve. These excitatory EPSCs were mediated by AMPA and N-methyl-D-aspartate (NMDA) receptors. The spontaneous EPSCs mediated by AMPA receptors had submillisecond decay kinetics (556 micros at E19), comparable with those of other auditory brain stem areas. The spontaneous EPSCs increased in amplitude and became faster with developmental age. Evoked EPSC and EPSP amplitudes were graded with stimulus intensity. The average amplitude of the EPSC evoked by minimal stimulation was twice as large as the average spontaneous EPSC amplitude (approximately 110 vs. approximately 55 pA), suggesting that single fibers make multiple contacts onto each postsynaptic NA neuron. Because of their small size, minimal EPSPs were subthreshold, and we estimate at least three to five inputs were required to reach threshold. In contrast to the fast EPSCs, EPSPs in NA had a decay time constant of approximately 12.5 ms, which was heavily influenced by the membrane time constant. Thus NA neurons spatially and temporally integrate auditory information arriving from multiple auditory nerve afferents.     

Synaptic organization of eighth nerve afferents to cat dorsal cochlear nucleus

The synaptic organization of eighth nerve afferents to the dorsal cochlear nucleus (DCN) of cats was studied using extracellular field potential analyses. The eighth nerve was electrically stimulated and potentials produced in the DCN characterized using single shocks, paired shocks, repetitive stimulation, and one-dimensional current source-density (CSD) analysis. Field potentials and CSD profiles were correlated with the laminated cytoarchitecture of the DCN. At least four major temporally discrete components can be identified in field potentials evoked by a single shock to the eighth nerve. The amplitude and polarity of these events depends on the layers in which they are recorded. A brief positive-negative deflection (the P1-N1) is present in all layers but is maximal in the deeper layers 3 and 4. The N1 has a peak latency of approximately 0.8 ms in these layers. The N1 in the deep layers is followed by a large negative potential, termed the N2, with a peak latency of about 1.6 ms. In the superficial layers (1 and 2), the N1 is followed by a small positive potential (the P2) occurring nearly simultaneously with the N2, Immediately following the N2 is another negative potential that is most clearly observed in layer 2. The layer 2 negative wave is termed N3 and is also identifiable on the repolarizing phase of the N2 in layers 3 and 4. The N3 in layer 2 can be followed by a positive potential, the P4. Simultaneous with the P4 is a small negative wave ion layer 1, termed the N4. The P4-N4 complex is observed in about half the recordings. Frequency-following tests indicate that both the N1 and N2 waves can follow shock trains up to 333 Hz. The N1 remains nearly constant in amplitude up to about 300 Hz and decreases as the stimulus frequency is raised to 500 Hz. The N2 decrements more rapidly than the N1 at frequencies above about 250 Hz and is considerably reduced at 500 Hz. The N2 sometimes shows an increased amplitude (by about 20-40%) between 100 and 250 Hz. A paired-shock paradigm was used to characterize further potentials. The N1 was little affected by prior stimulation for intervals greater than 5 ms. The N2 and N3 generally showed a small facilitation for shock intervals from about 7 to 30 ms, with a return to base line at longer intervals. The N4 and P4 (when present) were profoundly depressed for intervals from 7 to about 30 ms, with recovery to control values by 50 ms.(ABSTRACT TRUNCATED AT 400 WORDS)     

Synaptic and Golgi membrane recycling in cochlear hair cells

Summary Membrane recycling in the mechanoreceptive sensory cells of the mammalian cochlea was studied by observing membrane-bound horseradish peroxidase (HRP) reaction product following briefin vivo exposure to the enzyme. In the inner hair cell (IHC), peroxidase was taken up into coated vesicles and became incorporated into synaptic vesicles surrounding presynaptic bodies, but much HRP was also transported to the apical zone where reaction product appeared in all components of the Golgi complex. Neither the subsurface cisternae nor a tubular network associated with clusters of mitochondria were labelled. Outer hair cells (OHCs) showed considerably less membrane-bound reaction product than IHCs, indicating less rapid plasmalemmal recycling. Most membrane-bound reaction product was contained in coated vesicles and small vacuoles in the synaptic zone, but was occasionally seen in multivesicular bodies in the most apical zone. No labelled organelles were detected in the large central region of the OHC. A diffuse staining of the cytoplasm, particularly pronounced in OHCs, often interfered with the evaluation of membrane-bound reaction product in OHCs. This staining pattern could be qualitatively reproduced in both IHCs and OHCs by incubating fixed segments of the organ of Corti in oxidized diaminobenzidine. The presence of labelled synaptic vesicles associated with presynaptic bodies of IHCs and OHCs suggests that they are formed from membrane retrieved from the plasmalemma. We found no evidence that the subsurface cisternae of IHCs or the laminated cisternae of OHCs are derived from the cell surface as they never contained reaction product.     

Intracellularly labeled fusiform cells in dorsal cochlear nucleus of the gerbil. I. Physiological response properties

Fusiform cells in the dorsal cochlear nucleus (DCN) of barbiturate-anesthetized Mongolian gerbils were characterized physiologically and labeled with neurobiotin. This report is based on 17 fusiform cells for which there was reasonable confidence in the association between physiological data and recovered anatomy. The qualitative morphology of these cells was no different from that reported in previous studies. The acoustic response properties were generally consistent with those described in the barbiturate-anesthetized cat. Most responses were of the pauser or buildup type, but a dependence on stimulus frequency and intensity was observed. Stimulus-evoked sustained depolarizations and large, long-lasting afterhyperpolarizations were common membrane potential features. The cells in this study showed a greater tendency to discharge regularly than did those of the cat, likely as a result of the longer interstimulus interval used. Barbiturate anesthesia appears to mask an interspecies difference in DCN physiology that is apparent in unanesthetized, decerebrate preparations. The response of these fusiform cells to a depolarizing current pulse could be altered by the presence of a hyperpolarizing prepulse. Buildup, pauser, and chopper patterns could each be created using appropriate combinations of hyperpolarizing and depolarizing pulse amplitudes. Thus the adult gerbil appears to express the inactivating potassium conductance previously shown to affect fusiform cell firing patterns in vitro. The results further demonstrate that the effects of these potassium currents are readily observed in vivo. Finally, the fusiform cells in this study were quite variable with respect to a number of response properties, including the resting potential, input resistance, spontaneous activity, relative noise index, normalized tone slope, and regularity histogram shape. This diversity likely results from cell-to-cell variations in the balance of activity within the relatively complex network to which the fusiform cells belong, although effects of impalement may contribute to the extent of the diversity.     

Effects of OFF-BF tones on responses of chopper units in ventral cochlear nucleus. I. Regularity and temporal adaptation patterns.

1. We have recorded the responses of neurons in the anteroventral cochlear nucleus (AVCN) of barbiturate-anesthetized cats to pure tones [either at the unit's best frequency (BF) or at another frequency (OFF-BF)] and to two-tone combination stimuli. 2. The effects of OFF-BF input (either alone or presented simultaneously with a BF tone in a two-tone stimulus) on the response patterns of choppers may include not only rate inhibition but changes in the discharge regularity and the temporal adaptation properties of the spike trains. 3. In the majority of cases we studied (119 of 146 frequencies examined in 45 units), the discharge regularity of a response to an OFF-BF or two-tone stimulus is comparable with that of a "rate-matched" BF tone response. In a minority of cases (27 of 146 frequencies examined), however, OFF-BF input (either alone or in a two-tone stimulus format) changed the regularity compared with that of a rate-matched BF tone response. 4. In the majority of cases studied (139 of 171 frequencies examined in 53 units), the initial pattern of rate adaptation ["temporal adaptation pattern" (TAP)] was the same in response to a short tone burst at BF, to an OFF-BF tone burst, or to a pair of tones. The TAP can, however, be significantly altered by OFF-BF input, although this is a comparatively infrequent occurrence in our data sample (32 of 171 frequencies examined), from the response to BF tone to the response to the two-tone or OFF-BF stimulus, are as follows: sustained to slowly adapting; slowly adapting to transiently adapting, and transiently adapting to slowly adapting. Changes in the TAPs of chopper unit responses have been recorded from both regular and irregular choppers and cannot be accounted for on the basis of changes in sustained firing rate. These changes in the discharge regularity and TAP in the small minority of cases suggest that (at least in these cases) the inhibitory effect of OFF-BF input is not simply the result of two-tone suppression at the level of the auditory nerve fiber input. 5. We have observed that regular choppers may be transformed into irregular choppers by OFF-BF (rate inhibitory) input.(ABSTRACT TRUNCATED AT 400 WORDS)     

Laminar inputs from dorsal cochlear nucleus and ventral cochlear nucleus to the central nucleus of the inferior colliculus: two patterns of convergence

The central nucleus of the inferior colliculus is a laminated structure composed of oriented dendrites and similarly oriented afferent fibers that provide a substrate for tonotopic organization. Although inputs from many sources converge in the inferior colliculus, how axons from these sources contribute to the laminar pattern has remained unclear. Here, we investigated the axons from the cochlear nuclei that terminate in the central nucleus of the cat and rat. After characterization of the best frequency of the neurons at the injection sites in the cochlear nucleus, the neurons were labeled with dextran in order to visualize their axons and synaptic boutons in the central nucleus. Quantitative methods were used to determine the size and distribution of the boutons within the laminar organization. Two components in the laminae were identified: (1) a narrow axonal lamina that included the largest fibers and largest boutons; (2) a wide axonal lamina, surrounding the narrow lamina, composed of thin fibers and only small boutons. The wide lamina was approximately 30-40% wider than the narrow lamina, and it often extended more than 100 microm beyond the larger boutons on each side. The presence of both thick and thin fibers within the acoustic striae following these injections suggests that large and small fibers/boutons within these bands may originate from different neuronal types in the dorsal and ventral cochlear nucleus. We conclude that the narrow laminae that contain large fibers and boutons originate from larger cell types in the cochlear nucleus. In contrast, the wide lamina composed exclusively of small boutons may represent an input from other, perhaps smaller neurons in the cochlear nucleus. Thus, two types of inferior colliculus laminar structures may originate from the cochlear nucleus, and the small boutons in the wide laminae may contribute a functionally distinct input to the neurons of the inferior colliculus.     

Spontaneous discharge patterns in cochlear spiral ganglion cells before the onset of hearing in cats

Spontaneous neural activity has been recorded in the auditory nerve of cats as early as 2 days postnatal (P2), yet individual auditory neurons do not respond to ambient sound levels <90-100 dB SPL until about P10. Significant refinement of the central projections from the spiral ganglion to the cochlear nucleus occurs during this neonatal period. This refinement may be dependent on peripheral spontaneous discharge activity. We recorded from single spiral ganglion cells in kittens aged P3-P9. The spiral ganglion was accessed through the round window through the spiral lamina. A total of 112 ganglion cells were isolated for study in nine animals. Spike rates in neonates were very low, ranging from 0.06 to 56 spikes/s, with a mean of 3.09 +/- 8.24 spikes/s. Ganglion cells in neonatal kittens exhibited remarkable repetitive spontaneous bursting discharge patterns. The unusual patterns were evident in the large mean interval CV (CV(i) = 2.9 +/- 1.6) and burst index of 5.2 +/- 3.5 across ganglion cells. Spontaneous bursting patterns in these neonatal mammals were similar to those reported for cochlear ganglion cells of the embryonic chicken, suggesting this may be a general phenomenon that is common across animal classes. Rhythmic spontaneous discharge of retinal ganglion cells has been shown to be important in the development of central retinotopic projections and normal binocular vision. Bursting rhythms in cochlear ganglion cells may play a similar role in the auditory system during prehearing periods.     

Synaptic activity-induced Ca(2+) signaling in avian cochlear nucleus magnocellularis neurons

Neurons of the avian cochlear nucleus magnocellularis (NM) receive glutamatergic inputs from the spiral ganglion cells via the auditory nerve and feedback GABAergic inputs primarily from the superior olivary nucleus. We investigated regulation of Ca²⁺ signaling in NM neurons with ratiometric Ca²⁺ imaging in chicken brain slices. Application of exogenous glutamate or GABA increased the intracellular Ca²⁺ concentration ([Ca²⁺]_i) in NM neurons. Interestingly, GABA-induced Ca²⁺ responses persisted into neuronal maturation, in both standard and energy substrate enriched artificial cerebrospinal fluid. More importantly, we found that electrical stimulation applied to the glutamatergic and GABAergic afferent fibers innervating the NM was able to elicit transient [Ca²⁺]_i increases in NM neurons, and the amplitude of the Ca²⁺ responses increased with increasing frequency and duration of the electrical stimulation. Antagonists for ionotropic glutamate receptors significantly blocked these [Ca²⁺]_i increases, whereas blocking GABA_A receptors did not affect the Ca²⁺ responses, suggesting that synaptically released glutamate but not GABA induced the Ca²⁺ signaling in vitro. Furthermore, activation of GABA_A receptors with exogenous agonists inhibited synaptic activity-induced [Ca²⁺]_i increases in NM neurons, suggesting a role of GABA_A receptors in the regulation of Ca²⁺ homeostasis in the avian cochlear nucleus neurons.