Sound-induced seizures in serotonin 5-HT2c receptor mutant mice.

The epilepsies are a heterogeneous collection of seizure disorders with a lifetime expectancy risk rate of 2-4%. A convergence of evidence indicates that heritable factors contribute significantly to seizure susceptibility. Genetically epilepsy-prone rodent strains have been frequently used to examine the effect of genetic factors on seizure susceptibility. The most extensively studied of these have been strains that are susceptible to sound-induced convulsions (audiogenic seizures, or AGSs). Early observations of the AGS phenomenon were made in the laboratory of Dr. Ivan Pavlov; in the course of appetite-conditioning experiments in mice, the loud bell used to signal food presentation unexpectedly produced seizures in some animals. In 1947, DBA/2 (D2) mice were found to exhibit a genetic susceptibility to AGSs stimulated by a doorbell mounted in an iron tub. Since this discovery, AGSs have been among the most intensively studied phenotypes in behavioural genetics. Although several genetic loci confer susceptibility to AGSs, the corresponding genes have not been cloned. We report that null mutant mice lacking serotonin 5-HT2C receptors are extremely susceptible to AGSs. The onset of susceptibility is between two and three months of age, with complete penetrance in adult animals. AGS-induced immediate early gene expression indicates that AGSs are subcortical phenomena in auditory circuits. This AGS syndrome is the first produced by a known genetic defect; it provides a robust model for the examination of serotoninergic mechanisms in epilepsy.

Functional convergence of response properties in the auditory thalamocortical system.

One of the brain's fundamental tasks is to construct and transform representations of an animal's environment, yet few studies describe how individual neurons accomplish this. Our results from correlated pairs in the auditory thalamocortical system show that cortical excitatory receptive field regions can be directly inherited from thalamus, constructed from smaller inputs, and assembled by the cooperative activity of neuronal ensembles. The prevalence of functional thalamocortical connectivity is strictly governed by tonotopy, but connection strength is not. Finally, spectral and temporal modulation preferences in cortex may differ dramatically from the thalamic input. Our observations reveal a radical reconstruction of response properties from auditory thalamus to cortex, and illustrate how some properties are propagated with great fidelity while others are significantly transformed or generated intracortically.

Two thalamic pathways to primary auditory cortex.

Neurons in the center of cat primary auditory cortex (AI) respond to a narrow range of sound frequencies and the preferred frequencies in local neuron clusters are closely aligned in this central narrow bandwidth region (cNB). Response preferences to other input parameters, such as sound intensity and binaural interaction, vary within cNB; however, the source of this variability is unknown. Here we examined whether input to the cNB could arise from multiple, anatomically independent subregions in the ventral nucleus of the medial geniculate body (MGBv). Retrograde tracers injected into cNB labeled discontinuous clusters of neurons in the superior (sMGBv) and inferior (iMGBv) halves of the MGBv. Most labeled neurons were in the sMGBv and their density was greater, iMGBv somata were significantly larger. These findings suggest that cNB projection neurons in superior and iMGBv have distinct anatomic and possibly physiologic organization.

Unbalanced synaptic inhibition can create intensity-tuned auditory cortex neurons.

Intensity-tuned auditory cortex neurons have spike rates that are nonmonotonic functions of sound intensity: their spike rate initially increases and peaks as sound intensity is increased, then decreases as sound intensity is further increased. They are either "unbalanced," receiving disproportionally large synaptic inhibition at high sound intensities; or "balanced," receiving intensity-tuned synaptic excitation and identically tuned synaptic inhibition which neither creates enhances nor creates intensity-tuning. It has remained unknown if the synaptic inhibition received by unbalanced neurons enhances intensity-tuning already present in the synaptic excitation, or if it creates intensity-tuning that is not present in the synaptic excitation. Here we show, using in vivo whole cell recordings in pentobarbital-anesthetized rats, that in some unbalanced intensity-tuned auditory cortex neurons synaptic inhibition enhances the intensity-tuning; while in others it actually creates the intensity-tuning. The lack of balance between synaptic excitation and inhibition was not always apparent in their peak amplitudes, but could sometimes be revealed only by considering their relative timing. Since synaptic inhibition is essentially cortical in origin, the unbalanced neurons in which inhibition creates intensity-tuning provide examples of auditory feature-selectivity arising de novo at the auditory cortex.

Functional organization of the auditory cortex: maps and mechanisms.

Recent studies have led to a better understanding of several aspects of the organization and physiological mechanisms involved in the processing of information in the auditory cortex. A wide range of approaches have revealed new information regarding the histochemistry, cortico-cortical connections, single-unit physiology, and functional spatial organization, as well as mechanisms and effects of representational and learning-induced cortical plasticity.

Order and disorder in auditory cortical maps.

Recent physiological experiments suggest that several basic receptive field properties of neurons show non-uniform spatial distributions in the primary auditory cortex of cats and primates. The spatial distribution patterns of some of these receptive field parameters are suggestive of a parallel coding scheme for processing sound information onto several superimposed cortical 'maps'. The representations of these parameters in the auditory cortex are compatible with general features of self-organizing mapping algorithms, as the spatial representations exhibit global parameter gradients with overlaid functional patchiness. Recent studies have also revealed that within a 'representational' map, the degree of local coherence varies over a wide range and that the map can contain a substantial degree of disorder in its parametric representation of sounds. Although the causes and consequences of this representational disorder are not known, it may reflect yet unresolved organizational principles in the auditory cortex.

Functional congruity in local auditory cortical microcircuits.

Functional columns of primary auditory cortex (AI) are arranged in layers, each composed of highly connected fine-scale networks. The basic response properties and interactions within these local subnetworks have only begun to be assessed. We examined the functional diversity of neurons within the laminar microarchitecture of cat AI to determine the relationship of spectrotemporal processing between neighboring neurons. Neuronal activity was recorded across the cortical layers while presenting a dynamically modulated broadband noise. Spectrotemporal receptive fields (STRFs) and their nonlinear input/output functions (nonlinearities) were constructed for each neuron and compared for pairs of neurons simultaneously recorded at the same contact site. Properties of these local neuron pairs showed greater similarity than non-paired neurons within the same column for all considered parameters including firing rate, envelope-phase precision, preferred spectral and temporal modulation frequency, as well as for the threshold and transition of the response nonlinearity. This higher functional similarity of paired versus non-paired neurons was most apparent in infragranular neuron pairs, and less for local supragranular and granular pairs. The functional similarity of local paired neurons for firing rate, best temporal modulation frequency and two nonlinearity aspects was laminar dependent, with infragranular local pair-wise differences larger than for granular or supragranular layers. Synchronous spiking events between pairs of neurons revealed that simultaneous 'Bicellular' spikes, in addition to carrying higher stimulus information than non-synchronized spikes, encoded faster modulation frequencies. Bicellular functional differences to the best matched of the paired neurons could be substantial. Bicellular nonlinearities showed that synchronous spikes act to transmit stimulus information with higher fidelity and precision than non-synchronous spikes of the individual neurons, thus, likely enhancing stimulus feature selectivity in their target neurons. Overall, the well-correlated and temporally precise processing within local subnetworks of cat AI showed laminar-dependent functional diversity in spectrotemporal processing, despite high intra-columnar congruity in frequency preference.

Feature selectivity and interneuronal cooperation in the thalamocortical system.

Action potentials are a universal currency for fast information transfer in the nervous system, yet few studies address how some spikes carry more information than others. We focused on the transformation of sensory representations in the lemniscal (high-fidelity) auditory thalamocortical network. While stimulating with a complex sound, we recorded simultaneously from functionally connected cell pairs in the ventral medial geniculate body and primary auditory cortex. Thalamic action potentials that immediately preceded or potentially caused a cortical spike were more selective than the average thalamic spike for spectrotemporal stimulus features. This net improvement of thalamic signaling indicates that for some thalamic cells, spikes are not propagated through cortex independently but interact with other inputs onto the same target cell. We then developed a method to identify the spectrotemporal nature of these interactions and found that they could be cooperative or antagonistic to the average receptive field of the thalamic cell. The degree of cooperativity with the thalamic cell determined the increase in feature selectivity for potentially causal thalamic spikes. We therefore show how some thalamic spikes carry more receptive field information than average and how other inputs cooperate to constrain the information communicated through a cortical cell.

Strategies for optical control and simultaneous electrical readout of extended cortical circuits.

BACKGROUND: To dissect the intricate workings of neural circuits, it is essential to gain precise control over subsets of neurons while retaining the ability to monitor larger-scale circuit dynamics. This requires the ability to both evoke and record neural activity simultaneously with high spatial and temporal resolution. NEW METHOD: In this paper we present approaches that address this need by combining micro-electrocorticography (µECoG) with optogenetics in ways that avoid photovoltaic artifacts. RESULTS: We demonstrate that variations of this approach are broadly applicable across three commonly studied mammalian species - mouse, rat, and macaque monkey - and that the recorded µECoG signal shows complex spectral and spatio-temporal patterns in response to optical stimulation. COMPARISON WITH EXISTING METHODS: While optogenetics provides the ability to excite or inhibit neural subpopulations in a targeted fashion, large-scale recording of resulting neural activity remains challenging. Recent advances in optical physiology, such as genetically encoded Ca(2+) indicators, are promising but currently do not allow simultaneous recordings from extended cortical areas due to limitations in optical imaging hardware. CONCLUSIONS: We demonstrate techniques for the large-scale simultaneous interrogation of cortical circuits in three commonly used mammalian species.

Functional organization of spectral receptive fields in the primary auditory cortex of the owl monkey.

Recent experiments in the cat have demonstrated that several response parameters, including frequency tuning, intensity tuning, and FM selectivity, are spatially segregated across the isofrequency axis. To investigate whether a similar functional organization exists in the primate, we have studied the spatial distribution of pure-tone receptive field parameters across the primary auditory cortex (AI) in six owl monkeys (Aotus trivirgatus). The distributions of binaural interaction types and onset latency were also examined. Consistent with previous studies, the primary auditory cortex contained a clear cochleotopic organization. We demonstrate here that several other properties of the responses to tonal stimuli also showed nonrandom spatial distributions that were largely independent from each other. In particular, the sharpness of frequency tuning to pure tones, intensity tuning and sensitivity, response latency, and binaural interaction types all showed spatial variations that were independent from the representation of characteristic frequency and from each other. Statistical analysis confirmed that these organizations did not reflect random distributions. The overall organizational pattern of overlaying but independent functional maps that emerged was quite similar to that seen in AI of cats and, in general, appears to reflect a fundamental organization principle of primary sensory cortical fields.

Tonotopic and heterotopic projection systems in physiologically defined auditory cortex.

Combined physiological and connectional studies show significant non-topographic extrinsic projections to frequency-specific domains in the cat auditory cortex. These frequency-mismatched loci in the thalamus, ipsilateral cortex, and commissural system complement the predicted topographic and tonotopic projections. Two tonotopic areas, the primary auditory cortex (AI) and the anterior auditory field (AAF), were electrophysiologically characterized by their frequency organization. Next, either cholera toxin beta subunit or cholera toxin beta subunit gold conjugate was injected into frequency-matched locations in each area to reveal the projection pattern from the thalamus and cortex. Most retrograde labeling was found at tonotopically appropriate locations within a 1 mm-wide strip in the thalamus and a 2-3 mm-wide expanse of cortex (approximately 85%). However, approximately 13-30% of the neurons originated from frequency-mismatched locations far from their predicted positions in thalamic nuclei and cortical areas, respectively. We propose that these heterotopic projections satisfy at least three criteria that may be necessary to support the magnitude and character of plastic changes in physiological studies. First, they are found in the thalamus, ipsilateral and commissural cortex; since this reorganization could arise from any of these routes and may involve each, such projections ought to occur in them. Second, they originate from nuclei and areas with or without tonotopy; it is likely that plasticity is not exclusively shaped by spectral influences and not limited to cochleotopic regions. Finally, the projections are appropriate in magnitude and sign to plausibly support such rearrangements; given the rapidity of some aspects of plastic changes, they should be mediated by substantial existing connections. Alternative roles for these heterotopic projections are also considered.

Auditory neurophonic responses to amplitude-modulated tones: transfer functions and forward masking.

Two auditory neurophonic responses - one recorded from the scalp (frequency following response or FFR) and one from the auditory nerve (auditory nerve neurophonic or ANN) - were obtained following stimulation of the cat cochlea with amplitude-modulated (AM) high-frequency tones. The carrier frequencies varied between 2 and 30 kHz. The modulation frequencies varied between 400 and 3000 Hz. The AM responses were compared with pure-tone neurophonic responses. The AM response waveforms were found to have a similar spectral composition, similar rates of adaptation, and similar rates of recovery from forward masking as the comparable pure-tone responses. As with the pure-tone neurophonics, an unmodulated masking stimulus can produce prolonged depression of the probe response. The amount and duration of this depression is dependent upon the level and frequency of the masker. The frequency dependence of the depression is demonstrated by forward masked tuning curves which indicate that the AM responses arise from fiber populations which have restricted characteristic frequency distributions centered on the carrier frequency. Response amplitude as a function of stimulus level (I/O) functions, response amplitude as a function of carrier frequency (carrier transfer functions or CTF) and response amplitude as a function of modulation frequency (modulation transfer functions or MTF) were also measured. It was found that the I/O functions were saturating monotonic functions of stimulus intensity, CTFs were flat for carrier frequencies from 6 to 30 kHz, and MTFs were flat for modulation frequencies from 100 to 1500 Hz. These results are compared with similar data for single units and compound action potentials.

Representation of amplitude modulation in the auditory cortex of the cat. II. Comparison between cortical fields.

The responses of neuronal clusters to amplitude-modulated tones were studied in five auditory cortical fields of the anesthetized cat: the primary auditory field (AI), second auditory field (AII), anterior auditory field (AAF), posterior auditory field (PAF) and the ventro-posterior auditory field (VPAF). Modulation transfer functions (MTFs) for amplitude-modulated tones were obtained at 172 cortical locations. MTFs were constructed by measuring firing rate (rate-MTFs) and response synchronization (synchronization-MTFs) to sinusoidal and rectangular waveform modulation of CF-tones. The MTFs were characterized by their 'best-modulation frequency' (BMF) and a measure of their quality of 'sharpness' (Q2dB). These characteristics were compared for the five fields. Rate and synchronization MTFs for sinusoidal and rectangular modulation produced similar estimates of BMF and Q2dB. Comparison of averaged BMFs between the cortical fields revealed relatively high BMFs in AAF (mean: 31.1 Hz for synchronization to sinusoidal AM) and moderately high BMFs in AI (14.2 Hz) whereas BMFs encountered in AII, VPAF and PAF were generally low (7.0, 5.2, and 6.8 Hz). The MTFs were relatively broadly tuned (low Q2dB) in AAF and sharper in a low modulation group containing AII, PAF and VPAF. The ventro-posterior field was the most sensitive to changes in the modulation waveform. We conclude that there are significant differences between auditory cortical fields with respect to their temporal response characteristics and that the assessment of these response characteristics reveals important aspects of the functional significance of auditory cortical fields for the coding and representation of complex sounds.

The auditory neurophonic: basic properties.

In anesthetized cats an AC signal or neurophonic can be recorded from the auditory nerve and from the scalp when the cochlea is stimulated with low frequency tones. This study examines some of the basic properties of the auditory neurophonics. The auditory nerve signal, termed the auditory nerve neurophonic (ANN), was differentially recorded with a pair of platinum-iridium ball electrodes placed on either side of the auditory nerve as it exits the internal meatus. The signal recorded from the scalp, termed the frequency following response (FFR), was recorded with silver wire. For purposes of comparison the round window-recorded cochlear microphonic was also examined under identical stimulus conditions. Several measures of the response to acoustic stimulation were taken for each recording configuration. Among these were total response amplitude as a function of stimulus level, spectral component amplitude and phase as a function of stimulus level, fundamental component amplitude as a function of stimulus frequency, response amplitude as a function of time after stimulus onset, response amplitude as a function of forward masker intensity. By all these measures the neurophonic responses are signals that are distinct from the CM and share many of the properties of single units in the auditory nerve. In addition, micro-injections of kainic acid into the cochlear nucleus leave these responses largely unaffected, while tetrodotoxin injections into the cochlea greatly diminish both neurophonic responses, while leaving the CM largely intact. From these results, we conclude that at stimulus levels below 90 dB SPL the ANN is almost entirely neural in origin, while the FFR is certainly largely neural, that is, that both responses are quite distinct from the CM. We also conclude that they represent a spatial summation of neural activity in the auditory nerve, probably arising from the phase-locked response of single units to low frequency stimuli. In addition to demonstrating that the neurophonics are neural responses, we have begun the process of relating their properties to the distributed phase-locked activity in the auditory nerve.

Adaptation and recovery from adaptation of the auditory nerve neurophonic (ANN) using long duration tones.

The time course of the interaction between adaptation and the recovery from adaptation of the auditory nerve neurophonic (ANN) responses was examined. The interaction between the process of recovery and the adaptation process of long probe tones which follow a masker, the so called whole tone recovery, was determined for the ANN response for different silent intervals between masker offset and probe onset. The auditory nerve neurophonic (ANN) reflects the ensemble response of phase-locked firing in single auditory nerve fibers to sustained signals. Consequently, neural response properties such as adaptation and recovery from adaptation of these coherent, time-locked responses can be studied. Recovery from adaptation was determined by recording the response of a 290 ms duration probe tone following a 100 ms masker tone, equal in frequency to the probe, ranging from -5 to 20 dB relative to the probe amplitude. Two different time patterns of the whole tone recovery were observed. If short silent intervals and/or loud maskers were used, the time course of the probe tone can be described as an exponential increase in amplitude toward a steady state expressed by the equation: A(tp) = Ass-Yr e(-tp/tau Rr)-Ys e(-tp/tau Rs) ('ascending exponential'). For longer silent intervals and/or fainter maskers, the time course of the probe tone can be described by an exponential decrease expressed by the equation: A(tp) = Yr e(-tp/tau Rr) +Ys e(-tp/tau Rs) +A(ss) ('declining exponential').

Representation of amplitude modulation in the auditory cortex of the cat. I. The anterior auditory field (AAF).

The ability of cortical neurons to follow amplitude modulation (AM) of tones was examined in the anterior auditory cortical field (AAF) of anesthetized cats using multiple-unit recording techniques. Sinusoidal and rectangular modulations (100%) of a monaural carrier tone at the characteristic frequency of each location were presented to study the degree of response synchronization and changes in firing rate as a function of the modulation frequency. All investigated locations were tuned to a 'best modulation frequency' (BMF) as determined by synchronization measures. Almost all locations (94%) were tuned to a BMF as determined by spike rate. Maximal binaural-interaction strength was observed for modulation frequencies close to the BMF of neurons. For sinusoidal AM, a correlation (r = 0.63, P less than 0.01) between BMF and CF of neurons in AAF could be demonstrated for the synchronization of the response.

Forward masking of the auditory nerve neurophonic (ANN) and the frequency following response (FFR).

The forward masking behavior of two averaged neurophonic responses was examined in cats. The auditory nerve neurophonic (ANN) was recorded with bipolar electrodes placed on the auditory nerve as it exits the internal meatus. The frequency following response (FFR) was recorded using scalp electrodes placed at the vertex and below the stimulated ear. Masking functions (response amplitude vs masker level) for frequencies both above and below the probe frequency were recorded. From these masking functions, 30% iso-depression contours (forward masking tuning curves, FMTCs) were constructed. The time course of the recovery from forward masking was also examined. It was found that the forward masking behavior of these neurophonics have many similarities to the behavior of other responses recorded using psychophysical and physiological methods. However, forward masking of the ANN and FFR has a number of unusual features. First, the best masking frequency (BMF), which in most forward masking studies is equal to the probe frequency, can be off-set from the probe frequency by as much as an octave. Second, the masker level at BMF can be as much as 30 dB below the probe level. Third, the magnitude of both of these off-sets is a function of the probe level. Fourth, low level neurophonic response could be enhanced by some forward 'maskers'. The features of neurophonic forward masking are discussed and a model of the neurophonics is suggested. This model is based on the spatial distribution of phase and amplitude in the phase-locked activity in the auditory nerve and it can qualitatively account for many of the properties of the neurophonics.

Regional variations of noise-induced changes in operating range in cat AI.

Regional differences in spectral integration of neurons in cat primary auditory cortex (AI) suggest that regions differ in effects of background noise on operating characteristics of neurons. Therefore, tone-response threshold, best level (peak-rate intensity), dynamic range, and sharpness of tuning in quiet and in continuous broadband noise were mapped for single neurons along the isofrequency domain of AI. Neurons did not show an excitatory response to the noise. Noise invariably increased the tone-response threshold and best levels. Consequently, the dynamic ranges and receptive fields shifted to higher intensity levels without changes of average sharpness of tuning. These shifts were linearly related to noise level and showed little inter-neuronal variability for neurons in the central, mostly sharply tuned part of AI. In more dorsal and ventral parts of AI, neurons were more variable in tone-response threshold, dynamic range and best level, and no systematic relationship between increase in noise level, threshold increase and best-level increase was observed. We conclude that linear shifts in the operating range of neurons in central AI in the presence of continuous noise backgrounds do not affect other response properties and may relate to the unaltered analysis and representation of spectral components of sounds. In contrast, neurons in dorsal and ventral AI change response properties in a non-predictable way in the presence of noise in accordance with the more complex receptive field properties in those areas.

Effects of extracochlear direct current stimulation on the ensemble auditory nerve activity of cats.

The influence of direct current applied by round window stimulation on the whole nerve response of the auditory nerve of the cat has been studied. Effects on acoustically driven activity (CAP) and on the ensemble spontaneous activity of the nerve were observed. Stimulation with positive current suppressed driven and spontaneous activity. The strength and spread of suppressive effects was a function of the applied current level. After a period of positive electrical stimulation, driven and spontaneous activity rapidly returned to normal values. A rebound effect was sometimes observed, marked by a brief increase in spontaneous activity above the normal level. Negative current initially produced an increase in the amplitude of driven and spontaneous responses. Prolonged stimulation with negative current (greater than 30 s) resulted in a subsequent, graded reduction of neural activity, until a profound suppression of spontaneous and evoked neural activity was attained. The amplitude/latency relationship of CAPs was altered during passing of negative currents but not during passing of positive currents. Recovery from the suppression generated by negative currents was commonly prolonged for anything from a few seconds to many minutes; prolongation was dependent on stimulus amplitude, duration and duty cycles.

Auditory cortical neuron response differences under isoflurane versus pentobarbital anesthesia.

Response properties of the middle layers of feline primary auditory cortex neurons to simple sounds were compared for isoflurane versus pentobarbital anesthesia in a within subject study control design. Initial microelectrode recordings were made under isoflurane anesthesia. After a several hour washout period, recordings were repeated at spatially matched locations in the same animal under pentobarbital. The median spatial separation between matched recording locations was 50 microns. Excitatory frequency tuning curves (n=71 pairs) to tone bursts and entrainment to click train sequences (n=64 pairs) ranging from 2 to 38 Hz were measured. Characteristic frequency and BW10 and BW30 were not different under either anesthetic. The spontaneous rate was slightly decreased (P<0.05) for isoflurane (median 4.2 spikes/s) compared to pentobarbital (median 5.8 spikes/s). Minimum median threshold and latency were elevated by 12 dB and 2 ms, respectively, under isoflurane. Entrainment to click sequences assumed a lowpass filter profile under both anesthetics, but was markedly impoverished under isoflurane. Responses to click sequences under isoflurane were phasic to the first click but had very poor following to subsequent elements. Compared to pentobarbital, isoflurane appears to have a profound impact on response sensitivity and temporal response properties of auditory cortical neurons.