Different representations of tooth chatter and purr call in guinea pig auditory cortex

Multielectrode arrays were used to compare responses to tooth chatter and purr calls from all eight areas of the auditory cortex in anaesthetized guinea pigs. These calls have different behavioural contexts: males emit tooth chatters in aggressive encounters and the purr call during courtship behaviour. Of the two core areas, the primary auditory cortex responded better to both signals than the dorsocaudal core area. Of the six belt areas, the ventral transition area was found to be exceptionally sensitive to tooth chatter and less responsive to purr. The small rostral field responded faithfully to the purr, but not to tooth chatter, and ventrorostral belt often showed on/off responses; other belt areas were unresponsive.     

Dynamic spatiotemporal inhibition in the guinea pig auditory cortex

Real-time optical imaging was conducted in the guinea pig auditory cortex to study spatiotemporal interrelations of excitation and inhibition in response to tone stimulation. Tone stimulation elicited responses consisting of three phases in the anterior field (the primary auditory cortex of guinea pig) and in the dorsocaudal field of the auditory cortex. An early depolarization was followed by a late hyperpolarization and an even later depolarization both in the maximum excitatory regions and in the lateral regions beside and/or between them. The late hyperpolarization began significantly earlier and was stronger in the lateral regions than in the maximum excitatory regions. These results show that inhibition is dynamic, both in time and in space, in the auditory cortex.     

Phase-locked responses to pure tones in guinea pig auditory cortex

Phase-locked responses to pure tones are a characteristic of most auditory cells at the level of the brain stem and allow sophisticated analyses based on coincidence detection. Phase-locking to tones has not previously been shown at the level of the auditory cortex in single unit studies. We have now identified phase-locked responses in 10% of low-frequency (< 1 kHz) units in the ventrorostral belt, a strip of cortex immediately ventral to the primary auditory area. All of these units showed phase-locking in their response to binaural tone pips of 60-200 Hz and showed narrow band pass characteristics within this range.     

Functional subdivisions in the auditory cortex of the guinea pig

The auditory fields in the cortex of the guinea pig were investigated with microelectrode mapping techniques. Pure tones of varying frequencies and amplitudes were used as acoustic stimuli. Mainly, multiunit activity was recorded. A large tonotopic area is found in the anterior half of the auditory cortex. This area is named the anterior field (field A). Frequency tuning curves of multiunits in field A are generally narrow. Responses to tone stimuli are strong, and latencies are short. Low best frequencies are represented rostrally, high best frequencies caudally. The tonotopy is continuous and quite regular. Field A is narrow dorsally and becomes gradually broader ventrally. Correspondingly, the isofrequency lines slightly diverge from dorsal to ventral. Caudal to the first field, there is a second, smaller tonotopic area. It lies in the dorsal half of the posterior auditory cortex and is therefore named the dorsocaudal field (field DC). The frequency specificity of the cell clusters in this area is as strong as in field A, but the tonotopy is discontinuous: In the dorsal half of field DC, high best frequencies (16-32 kHz) are represented rostrally; the low frequencies (0.5-2.8 kHz) are represented immediately caudal to the high frequencies, while the intermediate frequencies are missing. Ventrally in field DC, the frequency representation is more complete. Except for this discontinuous map, we did not notice any differences between fields A and DC. A third tonotopic field was found rostral to field A. This field extends over a surface of less than 1 mm2 and was named the small field (field S). It contains a complete representation of the frequency range; high best frequencies are located rostrally, low frequencies caudally. The response latencies are slightly longer in field S than in fields A or DC, and the tuning curves are broader. A broad strip of nontonotopic cortex (auditory belt) surrounds fields A and DC caudally. We subdivided this area into the dorsocaudal and the ventrocaudal belt region. In both areas, tuning curves are often broad, and response latencies are longer than in the tonotopic cortex. In the dorsocaudal belt, most multiunits react with a phasic on-response to pure tones; in the ventrocaudal belt, tonic responses occur more frequently. Another nontonotopic region is located in the anterior auditory cortex, rostral to the tonotopic fields, and was therefore named the rostral belt. Tuning curves in this area are broad, latencies are short, and response thresholds are often high. In the discussion, the guinea pig is compared with other mammalian species.(ABSTRACT TRUNCATED AT 400 WORDS)     

A ventrorostral belt is adjacent to the guinea pig primary auditory cortex.

The previously defined anterior area (A) of guinea pig auditory cortex has been divided into a large dorsal portion identified as the primary area (AI) and a smaller ventrorostral belt (VRB). This division is based on: (1) the much longer response latency of units in VRB (21.7 ms) than AI (14.1 ms); (2) the absence of pure onset units in VRB (i.e. units that lacked a sustained response), which are common in AI; (3) the weakness of noise-induced evoked potentials in VRB compared to AI; (4) units in VRB had lower thresholds and stronger phase locking to amplitude modulated stimuli than in AI.     

Optical imaging of binaural interaction in multiple fields of the guinea pig auditory cortex

Locating the source of a sound is an important function of the auditory system and interaural intensity differences are one of the most important cues. To study the functional pathways of sound localisation processing in the auditory cortex, activity in multiple fields of the guinea pig auditory cortex during stimulation with interaural intensity differences was studied using optical imaging with a voltage-sensitive dye. Of the auditory core (primary and dorsocaudal) and the belt fields which surround them, the posterior and ventroposterior belt fields were the most sensitive to interaural intensity differences. This suggests that the caudal pathway of the auditory cortex is involved in sound localisation.     

Classical conditioning induces CS-specific receptive field plasticity in the auditory cortex of the guinea pig

To determine if classical conditioning produces general or specific modification of responses to acoustic conditioned stimuli (CS), frequency receptive fields (RF) of neurons in guinea pig auditory cortex were determined before and up to 24 h after fear conditioning. Highly specific RF plasticity characterized by maximal increased responses to the CS frequency and decreased responses to the pretraining best frequency (BF) and other frequencies was observed in 70% of conditioning cases. These opposing changes were often sufficient to produce a shift in tuning such that the frequency of the CS became the new BF. CS frequency specific plasticity was maintained as long as 24 h. Sensitization training produced general increased responses across the RF without CS specificity. The findings indicate that associative processes produce systematic modification of the auditory system's processing of frequency information and exemplify the advantages of combining receptive field analysis with behavioral training in the study of the neural bases of learning and memory.     

Sleep and wakefulness modulation of the neuronal firing in the auditory cortex of the guinea pig

Sleep-related changes—including modification in sensory processing—that influence brain and body functions, occur during both slow wave and paradoxical sleep. Our aim was to investigate how cortical auditory neurons behave during the sleep/waking cycle, and to study cell firing patterns in relation to the processing of auditory information without the interference of anesthetic drugs. We recorded single cells in the A region of the auditory cortex in restrained, chronically-implanted guinea pigs, and compared their evoked and spontaneous activity during sleep stages and quiet wakefulness. A new classification of the unit's responses to simple sound during wakefulness is presented. Moreover, a number of the neurons in the primary auditory cortex exhibited significant quantitative changes in their evoked or spontaneous firing rates. These changes could be correlated to sleep stages or wakefulness in 42.2% to 58.3% of the sampled neurons. A similar population did not show behavioral related changes in firing rates. Our results indicate that the responsiveness of the auditory system during sleep may be considered partially preserved. An important result was that spontaneous and evoked activity may vary in opposite directions, i.e., the evoked activity could increase while the spontaneous activity decrease or vice versa. Then, a general question was proposed: is the increased spontaneous activity in the auditory cortex, particularly during PS, related to auditory hypnic ‘images'? The studied cortical auditory neurons exhibit changes in their firing rates in correlation to stages of sleep and wakefulness. This is consistent with the hypothesis that a general shift in the neuronal networks involved in sensory processing occurs during sleep.     

Localization of acetylcholinesterase in the guinea pig cerebellar cortex

When using the acetylthiocholine method ofKoelle andGerebtzoff for the histochemical localization of acetylcholinesterase, a great variety of synapses within the cerebellar cortex of the guinea pig exhibit enzyme activity. A positive reaction was found in the cerebellar glomeruli of the granular layer, comprising both mossy and Golgi endings. Similar activity is present in the molecular layer, confined to parallel axons. A very strong enzyme reaction was found in the basket synapses around and below Purkinje cell bodies. The obviously presynaptic localization of acetylcholinesterase, and its presence in both excitatory and inhibitory synapses suggest that acetylcholine in these junctions plays the role of a pre-transmitter or modulator substance, promoting the activity of the very excitatory and inhibitory chemical transmitters.

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Zusammenfassung Eine beträchtliche Anzahl der Synapsen in der Kleinhirnrinde des Meerschweinchens zeigt eine Acetylcholinesterase-Aktivität, wenn formalinfixierte Gefrierschnitte nach der Acetylthiocholinmethode vonKoelle undGerebtzoff behandelt werden. In der Glomeruli cerebellares in der Körnerschichte sind Moosfasern und Golgi-Terminalen aktiv, während in der Molekularschicht das Enzym in den Parallelfasern lokalisiert ist. Eine sehr starke Enzymreaktion konnte in den Korbzellen-Synapsen um die Purkinjeschen Zellen nachgewiesen werden. Die präsynaptische Lokalisation von Acetylcholinesterase, und das Vorhandensein dieses Enzyms in funktionell antagonistischen erregenden und hemmenden Synapsen läßt vermuten, daß das Acetylcholin in diesen Synapsen die Rolle einer Modulator-Substanz spielt, wodurch die tatsächlich erregenden oder hemmenden Transmitter-Substanzen ihre reizübertragende Wirkung ausüben können.

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Résumé La technique deKoelle etGerebtzoff pour la localisation histochimique d'acétylcholinestérase montre une activité enzymatique dans différentes synapses cérébelleuses du cobaye à l'aide de coupes à congélation après fixation formalinique. On a trouvé une réaction positive dans les glomérules cérébelleux, ainsi dans les fibres mousseuses et dans les terminaisons de cellules de Golgi. Une activité également positive est présente dans la couche moléculaire dans les axons parallèles. Une activité très forte se trouve dans les synapses en forme de corbeilles des cellules de Purkinje. La localisation de l'acétylcholinestérase est évidemment présynaptique. La présence de l'enzyme dans les synapses excitatives et inhibitrices suggère que l'acétylcholine joue rôle d'une substance modulatrice, par laquelle les substances excitatives ou inhibitrices de transmission peuvent exercer leur efficacité.

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With 6 Figures     

Biphasic changes in tissue partial pressure of oxygen closely related to localized neural activity in guinea pig auditory cortex.

An understanding of the local changes in cerebral oxygen content accompanying functional brain activation is critical for making a valid signal interpretation of hemodynamic-based functional brain imaging. However, spatiotemporal relations between changes in tissue partial pressure of oxygen (Po2) and induced neural activity remain incompletely understood. To characterize the local Po2 response to the given neural activity, the authors simultaneously measured tissue Po2 and neural activity in the identical region of guinea pig auditory cortex with an oxygen microelectrode (tip < 10 microm) and optical recording with voltage-sensitive dye (RH 795). In addition, a laser displacement gauge and a laser-Doppler flowmeter were used to monitor the spatial displacement and regional cerebral blood flow, respectively, in the Po2 measurement region. In the activated region, tissue Po2 initially decreased during the approximately 3-seconds after the onset of acoustic stimuli, and then increased during the next approximately 5 seconds. Such biphasic changes are consistently found in cortical layers I to IV. In addition, amplitude of the biphasic change was closely related to detected peak height of the optical signal changes. The results suggest that the initial decrease in tissue Po2 is coupled to the induced neural activity and depends on response time of local increase in cerebral blood flow.     

Projections from the auditory cortex to the superior olivary complex in guinea pigs

We used anterograde tracing techniques to characterize projections from auditory cortex to the superior olivary complex (SOC) in guinea pigs. Large injections of fluorescent or biotinylated dextrans into the temporal cortex labeled many axons in the SOC. Labeled boutons were most numerous in the ventral nucleus of the trapezoid body, with additional boutons in all other olivary nuclei. The distribution of boutons was similar in the ipsilateral and contralateral SOC; however, the contralateral SOC had markedly fewer axons and boutons. Similar patterns of labeling were also observed following injections confined to primary auditory cortex or the dorsocaudal auditory field. Cortical axons in many of the SOC nuclei share numerous morphological features, suggesting that individual axons may innervate multiple nuclei and have widespread effects. In addition, some nuclei contain axons with branching or termination patterns unique to that nucleus; these axons may represent focused projections with effects limited to individual SOC nuclei. Given the many projections of SOC nuclei, cortico-olivary projections are in a position to modify the activity of many brainstem auditory circuits.     

Anatomy of the auditory thalamocortical system of the guinea pig

We investigated the projection from the medial geniculate body (MG) to the tonotopic fields (the anterior field A, the dorsocaudal field DC, the small field S) and to the nontonotopic ventrocaudal belt in the auditory cortex of the guinea pig. The auditory fields were first delimited in electrophysiological experiments with microelectrode mapping techniques. Then, small quantities of horseradish peroxidase (HRP) and/or fluorescent retrograde tracers were injected into the sites of interest, and the thalamus was checked for labeled cells. The anterior field A receives its main thalamic input from the ventral nucleus of the MG (MGv). The projection is topographically organized. Roughly, the caudal part of the MGv innervates the rostral part of field A and vice versa. After injection of tracer into low or medium best-frequency sites in A, we also found a topographic gradient along the isofrequency contours: the dorsal (ventral) part of a cortical isofrequency strip receives afferents from the rostral (caudal) portions of the corresponding thalamic isofrequency band. However, it is not so obvious whether such a gradient exists also in the high-frequency part of the projection. A second, weaker projection to field A originates in a magnocellular nucleus that is situated caudomedially in the MG and was therefore named the caudomedial nucleus. The dorsocaudal field DC receives input from the same nuclei as the anterior field, but the location of the labeled cells in the MGv is different. This was demonstrated by injection of different tracers into sites with like best frequencies in fields A and DC, respectively. After injection of HRP into the 1-2-kHz isofrequency strip in field A and injection of Nuclear Yellow (NY) into the 1-2-kHz site in field DC, the labeled cells in the MGv form one continuous array that runs from caudal to rostral over the whole extent of the MGv. The anterior part of this array consists of NY-labeled cells; i.e., it projects to field DC. The caudal part is formed by HRP-labeled cells; i.e., it innervates field A. These findings indicate that there is only one continuous tonotopic map in the MGv. This map is split when projected onto the cortex so that two adjacent tonotopic fields (A and DC) result. The cortical maps are rotated relative to the thalamic map in that rostral portions of the MGv project to caudal parts of the tonotopic cortex and vice versa.(ABSTRACT TRUNCATED AT 400 WORDS)     

Ipsilateral projection from the subiculum to the retrosplenial cortex in the guinea pig

A projection from the subiculum to the retrosplenial cortex (area 29a and b) was studied by autoradiographic tracing of anterogradely transported proteins following injections of radioactive amino acids into ventral and dorsal parts of the subiculum. Only the dorsal parts of the subiculum projected to the retrosplenial cortex. The projection was exclusively ipsilateral, and topographically arranged along both the longitudinal septotemporal axis and the transverse plane of the subiculum. The termination in the retrosplenial cortex was confined to the relatively cell free superficial layer (lamina 1) and the underlying cell-rich layers (laminae 3–4). Between these two terminations a narrow zone (lamina 2) was only weakly labeled. The projection constitutes an output route for hippocampal activity in addition to the fimbria-fornix system and the subicular projections to the parahippocampal areas.     

Connections of functional areas in the mustached bat's auditory cortex with the auditory thalamus

The auditory thalamus is the major target of the inferior colliculus and connects in turn with the auditory cortex. In the mustached bat, biosonar information is represented according to frequency in the central nucleus of the inferior colliculus (ICc) but according to response type in the cortex. In addition, the cortex has multiple areas with neurons of similar response type compared to the single tonotopic representation in the ICc. To investigate whether these transformations occur at the level of the thalamus, we injected anatomical tracers into physiologically defined locations in the mustached bat's auditory cortex. Injections in areas used for target ranging labeled contiguous regions of the auditory thalamus rather than separate patches corresponding to regions that respond to the different harmonic frequencies used for ranging. Injections in the two largest ranging areas produced labeling in separate locations. These results indicate that the thalamus is organized according to response type rather than frequency and that multiple mappings of response types exist. Injections in areas used for target detection labeled thalamic regions that were largely separate from those that interconnect with ranging areas. However, injections in an area used for determining target velocity overlapped with the areas connected to ranging areas and areas involved in target detection. Thus, separation by functional type and multiplication of areas with similar response type occurs by the thalamic level, but connections with the cortex segregate the functional types more completely than occurs in the thalamus.     

Localization of lipoprotein lipase to discrete areas of the guinea pig brain

Lipoprotein lipase is a key enzyme in lipoprotein metabolism present primarily in extrahepatic tissues with high turnover of fatty acids. Using immunocytochemistry we have explored where lipoprotein lipase is localized in guinea pig brain. The enzyme was found to be associated with neuronal cells and vascular endothelial surfaces. The distribution was strikingly uneven with intense reaction in some areas, and virtually no reaction in adjacent areas. The highest reactivity was in neocortex, in hippocampus, in Purkinje cells of the cerebellum and in some motor nuclei of the brainstem. The results suggest marked differences between individual brain areas in utilization of plasma lipoproteins.     

Acetylcholine modifies neuronal acoustic rate-level functions in guinea pig auditory cortex by an action at muscarinic receptors

Cholinergic modification of neuronal responsiveness in auditory cortex includes alteration of spontaneous and tone-evoked neuronal discharge. Previously it was suggested that the effects of acetylcholine (ACh) and muscarinic agonists on neuronal discharge resembled those due to increases in the intensity of acoustic stimuli (Ashe et al. 1989). To determine the relationship between neuronal modifications due to ACh acting at muscarinic receptors and those due to changes in stimulus intensity, we determined acoustic rate-level functions for neurons in the auditory cortex of barbiturate-anesthetized guinea pigs before, during and after administration of ACh. ACh facilitated acoustic rate-level functions in 82% of the cells tested. In addition, during ACh administration 66% of neurons responded to stimuli that were previously subthreshold, that is, ACh decreased the response threshold. Cholinergic facilitation of rate-level functions was attenuated by the general muscarinic antagonist atropine. The nature of the muscarinic receptors involved in the actions of ACh was further examined by presenting single tones before, during, and after administration of ACh and specific muscarinic receptor subtype antagonists, either pirenzepine (M1) or gallamine (M2). ACh-induced facilitation of spontaneous and tone evoked neuronal discharge was antagonized by pirenzepine, but not by gallamine, suggesting the involvement of the M1 muscarinic receptor subtype. These data indicate that ACh can facilitate stimulus-evoked responses and decrease response thresholds for neurons in auditory cortex, possibly via activation of M1 muscarinic receptors. Such effects of ACh acting at muscarinic receptors could underly cholinergic regulation of information processing in the auditory cortex.     

Projections from auditory cortex to cholinergic cells in the midbrain tegmentum of guinea pigs

Anterograde and retrograde tracing techniques were used to characterize projections from the auditory cortex to the pedunculopontine and laterodorsal tegmental nuclei (PPT and LDT, respectively) in the midbrain tegmentum in guinea pigs. For anterograde tracing, tetramethylrhodamine dextran (FluoroRuby) was injected at several sites within auditory cortex. After sufficient time for transport, the brain was processed for immunohistochemistry with anti-choline acetyltransferase to reveal presumptive cholinergic cells. Anterogradely labeled axons were observed ipsilaterally and, in smaller numbers, contralaterally, in both the pedunculopontine and laterodorsal tegmental nuclei. In all four nuclei, tracer-labeled boutons appeared to contact immunolabeled (i.e., cholinergic) cells. The contacts occurred on cell bodies and dendrites. The results were similar following injections that spread across multiple auditory cortical areas or injections that were within primary auditory cortex. In order to confirm the anterograde results, in a second series of experiments, retrograde tracers were deposited in the pedunculopontine tegmental nucleus. These injections labeled layer V pyramidal cells in the auditory cortex. The results suggest an excitatory projection from primary auditory cortex bilaterally to cholinergic cells in the midbrain tegmentum. Such a pathway could allow auditory cortex to activate brainstem cholinergic circuits, possibly including the cholinergic pathways associated with arousal and gating of acoustic stimuli.     

Spatio-temporal analyses of stimulus-evoked and spontaneous stochastic neural activity observed by optical imaging in guinea pig auditory cortex

Stimulus-evoked response in the cortex involves random neural activity besides the deterministic responses reproducible to the stimulus. Recently, we have developed a new bright optical system that enables us to investigate the spatio-temporal patterns of such stochastic activity in the guinea pig auditory cortex without averaging. We show that (1) the stochastic neural activity is evoked by a tone-stimulus in addition to the deterministic response, and spontaneous stochastic activity is also observed in a similar manner; (2) our statistical estimation of optical responses such as variance showed that the evoked stochastic activity was increased by the sound stimulus compared to the spontaneous activity; (3) both types of stochastic activity mainly display oscillatory behavior, in the frequency range of 5-11 Hz; (4) there are no significant differences between the stimulus-induced and spontaneous stochastic neural activity in our statistical analyses using the PSD (power-spectrum density) and the spatial correlation function; (5) the spatial area of the evoked stochastic activity is not strongly correlated with the tonotopical area of the deterministic response that is mainly localized in the caudal area of field A of the guinea pig auditory cortex. Thus, the stochastic neural activity existing in the stimulus response and the spontaneous activity in the auditory cortex are possibly generated by a common neural mechanism. These results were confirmed statistically using 27 animals.     

Responses to linear and logarithmic frequency-modulated sweeps in ferret primary auditory cortex

Multi-unit responses to frequency-modulated (FM) sweeps were studied in the primary auditory cortex of ferrets using six different stimulation paradigms. In particular, the differences between the responses to linear FM sweeps (where frequency changes linearly with time) and logarithmic FM sweeps (where frequency changes exponentially with time) were emphasized. Some general features of the responses to FM sweeps are independent of the exact details of the frequency trajectory. Both for linear and for logarithmic FM sweeps, a short burst of spikes occurred when the sweep reached a triggering frequency close to the best frequency of the cluster. The neuronal preference for FM velocity was also independent of frequency trajectory. Thus, clusters that responded best to slow logarithmic FM also preferred slow linear FM and vice versa. Consequently, topographic distributions of velocity preference were roughly independent of the stimulation paradigm. Other characteristics of the responses, however, depended on the exact details of the frequency trajectory. A significant number of clusters showed large differences in directional sensitivity between linear and logarithmic FM sweeps; these differences depended on the velocity preference of the clusters in some paradigms but not in others. Consequently, topographic distributions of directional sensitivity differed between linear and logarithmic paradigms. In conclusion, some characteristics of cluster responses to FM sweeps depend on the exact details of the stimulation paradigm and are not 'invariants' of the cluster.