Excitatory Area Research Articles

A brainstem circuit for nausea suppression.

SUMMARYNausea is a discomforting sensation of gut malaise that remains a major clinical challenge. Several visceral poisons induce nausea through the area postrema, a sensory circumventricular organ that detects blood-borne factors. Here, we use genetic approaches based on an area postrema cell atlas to reveal inhibitory neurons that counteract nausea-associated poison responses. The gut hormone glucose insulinotropic peptide (GIP) activates area postrema inhibitory neurons that project locally and elicit inhibitory currents in nausea-promoting excitatory neurons through γ-aminobutyric acid (GABA) receptors. Moreover, GIP blocks behavioral responses to poisons in wild-type mice, with protection eliminated by targeted area postrema neuron ablation. These findings provide insights into the basic organization of nausea-associated brainstem circuits and reveal that area postrema inhibitory neurons are an effective pharmacological target for nausea intervention.

Frequency response areas of neurons in the mouse inferior colliculus. III. Time-domain responses: Constancy, dynamics, and precision in relation to spectral resolution, and perception in the time domain.

The auditory midbrain (central nucleus of inferior colliculus, ICC) receives multiple brainstem projections and recodes auditory information for perception in higher centers. Many neural response characteristics are represented in gradients (maps) in the three-dimensional ICC space. Map overlap suggests that neurons, depending on their ICC location, encode information in several domains simultaneously by different aspects of their responses. Thus, interdependence of coding, e.g. in spectral and temporal domains, seems to be a general ICC principle. Studies on covariation of response properties and possible impact on sound perception are, however, rare. Here, we evaluated tone-evoked single neuron activity from the mouse ICC and compared shapes of excitatory frequency-response areas (including strength and shape of inhibition within and around the excitatory area; classes I, II, III) with types of temporal response patterns and first-spike response latencies. Analyses showed covariation of sharpness of frequency tuning with constancy and precision of responding to tone onsets. Highest precision (first-spike latency jitter < 1 ms) and stable phasic responses throughout frequency-response areas were the quality mainly of class III neurons with broad frequency tuning, least influenced by inhibition. Class II neurons with narrow frequency tuning and dominating inhibitory influence were unsuitable for time domain coding with high precision. The ICC center seems specialized rather for high spectral resolution (class II presence), lateral parts for constantly precise responding to sound onsets (class III presence). Further, the variation of tone-response latencies in the frequency-response areas of individual neurons with phasic, tonic, phasic-tonic, or pauser responses gave rise to the definition of a core area, which represented a time window of about 20 ms from tone onset for tone-onset responding of the whole ICC. This time window corresponds to the roughly 20 ms shortest time interval that was found critical in several auditory perceptual tasks in humans and mice.

Experimental tooth movement temporally changes neural excitation and topographical map in rat somatosensory cortex

During orthodontic treatment, binding teeth, may change the topographically organized representation of teeth in the cerebral cortex. To test the hypothesis that experimental tooth movement (ETM) changes the somatotopy of an individual tooth arrangement in the somatosensory cortex, we examined the spatiotemporal features of cortical excitatory propagation in response to mechanical stimulation of the maxillary incisor or molar using optical imaging in late adolescent rats without or with ETM. The ETM models consisted of 1d, 3d, and 7d ETM in which a closed-coil spring was ligated between the maxillary first molar and incisors. In controls, incisor and molar mechanical stimulation evoked excitation in the rostral and dorsocaudal regions of the primary somatosensory cortex (S1), respectively. In addition, the secondary somatosensory cortex and insular oral region (S2/IOR) were also activated. Incisor stimulation-induced excitatory regions in S1 of 3d and 7d ETM shifted without changing the maximum excitatory area or peak amplitude; the incisor stimulation-responding region moved toward the dorsocaudal region, which responded to molar stimulation in the control. This shift in excitatory region was not observed in 1d ETM. One day after removal of the coil spring that was attached for 6 days, the excitatory region shift in S1 was recovered to the control region. On the other hand, 1d ETM exhibited facilitation of the excitatory area and peak amplitude upon molar stimulation, and the facilitation of excitatory propagation disappeared in 3d and 7d ETM. These results may explain the clinical finding that abnormal sensation temporally occurs during orthodontic treatment.

Spatiotemporal profiles of dental pulp nociception in rat cerebral cortex: an optical imaging study.

Somatosensation is topographically organized in the primary (S1) and secondary somatosensory cortex (S2), which contributes to identify the region receiving sensory inputs. However, it is still unknown how somatosensory inputs from the oral region, especially nociceptive inputs from the teeth, are processed in the somatosensory cortex. We performed in vivo optical imaging and identified the precise cortical regions responding to electrical stimulation of the maxillary and mandibular dental pulp in rats. Electrical stimulation of the mandibular incisor pulp evoked neural excitation in two areas: the most rostroventral part of S1, and the ventral part of S2 caudal to the middle cerebral artery. Maxillary incisor pulp stimulation initially evoked responses only in the ventral part of S2, although later maximum responses were also observed in S1 similar to mandibular incisor stimulation responses. The maxillary and mandibular molar pulp-responding regions were located in the most ventral S2, a part of which was histologically classified as the insular oral region (IOR). In terms of the initial responses, maxillary incisor and molar stimulation induced excitation in the S2/IOR rostral to the mandibular dental pulp-responding region. Contrary to the spatially segregated initial responses, the maximum excitatory areas responding to both incisors and molars in the mandible and maxilla overlapped in S1 and the S2/IOR. Multielectrode extracellular recording supported the characteristic localization of S2/IOR neurons responding to mandibular and maxillary molar pulp stimulation. The discrete and overlapped spatial profiles of initial and maximum responses, respectively, may characterize nociceptive information processing of dental pain in the cortex.

Biosonar navigation above water II: exploiting mirror images.

As in vision, acoustic signals can be reflected by a smooth surface creating an acoustic mirror image. Water bodies represent the only naturally occurring horizontal and acoustically smooth surfaces. Echolocating bats flying over smooth water bodies encounter echo-acoustic mirror images of objects above the surface. Here, we combined an electrophysiological approach with a behavioral experimental paradigm to investigate whether bats can exploit echo-acoustic mirror images for navigation and how these mirrorlike echo-acoustic cues are encoded in their auditory cortex. In an obstacle-avoidance task where the obstacles could only be detected via their echo-acoustic mirror images, most bats spontaneously exploited these cues for navigation. Sonar ensonifications along the bats' flight path revealed conspicuous changes of the reflection patterns with slightly increased target strengths at relatively long echo delays corresponding to the longer acoustic paths from the mirrored obstacles. Recordings of cortical spatiotemporal response maps (STRMs) describe the tuning of a unit across the dimensions of elevation and time. The majority of cortical single and multiunits showed a special spatiotemporal pattern of excitatory areas in their STRM indicating a preference for echoes with (relative to the setup dimensions) long delays and, interestingly, from low elevations. This neural preference could effectively encode a reflection pattern as it would be perceived by an echolocating bat detecting an object mirrored from below. The current study provides both behavioral and neurophysiological evidence that echo-acoustic mirror images can be exploited by bats for obstacle avoidance. This capability effectively supports echo-acoustic navigation in highly cluttered natural habitats.

Brief exposure of juvenile rats to noise impairs the development of the response properties of inferior colliculus neurons

Temporary impairment of the auditory periphery during the sensitive period of postnatal development of rats may result in a deterioration of neuronal responsiveness in the central auditory nuclei of adult animals. In this study, juvenile rats (postnatal day 14) were exposed for 8 min to intense broad-band noise; at the age of 3-6 months, the excitatory and inhibitory response areas of neurons in the central nucleus of the inferior colliculus were recorded under ketamine-xylazine anaesthesia in these animals and compared with those of age-matched controls. The response thresholds were similar in the exposed and control animals. The frequency selectivity of low-frequency neurons was comparable in both groups; however, high-frequency neurons had significantly wider excitatory response areas in the exposed rats, indicating disrupted development of high-frequency hearing. Forty-one per cent and 25% of neurons in exposed animals and in controls, respectively, lacked a distinct inhibitory area; these neurons had similar frequency selectivity in the exposed and control rats. As the presence of an inhibitory sideband was associated with sharper frequency tuning in both groups, it appears that lateral inhibition substantially influences neuronal frequency selectivity. If present, the inhibitory areas had comparable bandwidths in both groups; however, they were shifted to the side in the exposed animals, allowing the expansion of the excitatory areas. The results indicate that a brief exposure of juvenile rats to noise leads to a significant worsening of the frequency selectivity of inferior colliculus neurons in adult animals; the poorer frequency selectivity may be due to missing or displaced inhibitory sidebands.

Extraction of visual motion and optic flow

In an earlier paper [Fukushima, K., & Tohyama, K. (2005). Analysis of optic flow: a neural network model. In ICONIP 2005, International Conference on Neural Information Processing (pp. 312-317); Tohyama, K., & Fukushima, K. (2005). Neural network model for extracting 98 optic flow. Neural Networks, 18(5-6), 549-556], we proposed a neural network extracting optic flow based on vector field hypothesis. We started discussion there, however, from the stage where local velocities of visual objects have already been extracted. This paper proposes a new mechanism of extracting local velocity from retinal images, and adds it to the previous network. The network has a hierarchical multilayered architecture. X- and Y-cells of the retina extract spatial and temporal contrast of brightness. The network contains several types of V1 cells, namely, S- C- and V-cells. S- and C-cells extract orientated edges. V-cells extract local velocities, based on the signals from Y- and S-cells. MT cells extract relative velocities between adjoining small visual fields. MST cells add the responses of MT cells to extract a specific optic flow, such as rotation and expansion/contraction. The difference in type of optic flow extracted by MST cells can be created simply by the difference in relative locations of inhibitory to excitatory areas in the receptive fields of MT cells.

Response Properties of Single Units in the Dorsal Nucleus of the Lateral Lemniscus of Decerebrate Cats

The dorsal nucleus of the lateral lemniscus (DNLL) receives afferent inputs from many brain stem nuclei and, in turn, is a major source of inhibitory inputs to the inferior colliculus (IC). The goal of this study was to characterize the monaural and binaural response properties of neurons in the DNLL of unanesthetized decerebrate cat. Monaural responses were classified according to the patterns of excitation and inhibition observed in contralateral and ipsilateral frequency response maps. Binaural classification was based on unit sensitivity to interaural level differences. The results show that units in the DNLL can be grouped into three distinct types. Type v units produce contralateral response maps that show a wide V-shaped excitatory area and no inhibition. These units receive ipsilateral excitation and exhibit binaural facilitation. The contralateral maps of type i units show a more restricted I-shaped region of excitation that is flanked by inhibition. Type o maps display an O-shaped island of excitation at low stimulus levels that is bounded by inhibition at higher levels. Both type i and type o units receive ipsilateral inhibition and exhibit binaural inhibition. Units that produce type v maps have a low best frequency (BF), whereas type i and type o units have high BFs. Type v and type i units give monotonic rate-level responses for both BF tones and broadband noise. Type o units are inhibited by tones at high levels, but are excited by high-level noise. These results show that the DNLL can exert strong, differential effects in the IC.

Analysis of Spatio-Temporal Neural Activities by Artificial Neural Network

The artificial neural network (ANN) can reconstruct spatio-temporal neural activities into the corresponding test stimuli. ANN with a simple structure and generalization ability has a potential to reflect a prominent feature of the mechanism of neural computation in the brain. In the present work, we test this hypothesis and propose a novel analysis by investigating input-output relationships of hidden layer neurons. We made ANN with neural activities in the primary auditory cortex serving as the inputs and time-series changes of test frequencies of tones serving as the targets. We then investigated the hidden layer neurons that played important roles in the reconstruction. Neurons that tuned the frequency preference by excitatory inputs had positive contribution from all frequency regions. On the other hand, neurons responsible for inhibitory frequency tuning had negative contribution from a low frequency region. These results suggest that neural activities in the primary auditory cortex form a frequency preference with excitatory inputs from all frequency pathways and inhibitory inputs from a low frequency pathway. This suggestion is consistent with physiological facts that pyramidal cells in the auditory cortex have widely tuned excitatory response area and inhibitory input domains that flank the excitatory areas, supporting our hypothesis and proving the feasibility of the proposed analysis.

Respiratory Effects of Stimulation of the Limbic Cortex in Rats and Their Modulation with Serotonin

Acute experiments on rats showed that the anterior cingulate gyrus contains 2 efferent regions of the functional respiratory system (inhibitory supragenual and excitatory infragenual areas). Stimulation of these cortical areas produced a respiratory effect, which depended on activity of serotoninergic mechanisms in the solitary tract nucleus.

Changes in the Latency of Mouse Inferior Colliculus Neuron Responses Depending on the Position and Direction of Movement of Spectral Contrast

Changes in the latency of mouse (Mus musculus) inferior colliculus neuron responses in the presence of wideband sound signals with spectral notches and noise bands with regularly varying central notch/band frequencies were studied. Relationships between the latency and the magnitude of the response on the one hand and the central notch/band frequency on the other were obtained (latency and spike count functions). Crossing of the margins of the excitatory areas of the responses of the frequency receptive fields of neurons by spectral notches/noise bands could lead to displacement of latency functions (and corresponding displacements in spike count functions). Direction-dependent shifts in latency and spike count functions were more characteristic of primary-like and V-shaped neurons. The most interesting feature of the directional sensitivity of inhibition-dependent neurons was the selective decrease in the latency and selective synchronization of the initial spike response (with a corresponding increase in the spike count). The dynamic properties of inhibition-dependent neurons can be explained on the basis of their selective sensitivity to the position of the spectral contrast in the frequency receptive field, which is associated with disinhibition, and by the nature of the distribution of the excitatory and inhibitory inputs. The extents of these effects depended on the spectral shape of the signals and the widths of the spectral notches.

Activity of Neurons in the Mouse Inferior Colliculus in Relation to the Position and Direction of Displacement of Spectral Contrast

The variability of the responses of neurons in the inferior colliculus of the mouse (Mus musculus) to sequences of signals of noise bands and wide-band noise with spectral notches with regular changes (by 1/12 octave) in the central band frequency or notch frequency was studied. When the width of spectral changes was 1/3 octave, neurons with strong inhibitory influences in the excitatory zone of the response ("inhibition-dependent" neurons) showed low levels of spike activity if the noise band completely covered the excitatory part of the response. The most effective stimuli for these neurons were spectral contrasts passing through the center of the excitatory part of the response (through or close to the characteristic frequency). The responses of neurons to spectral contrasts created by noise bands and noises with notches were identical. It is suggested that approximation of the inhibitory and excitatory inputs sharpens the frequency tuning of neurons to the position of the spectral contrasts, as occurs in the visual system. The selectivity of neurons to the direction of moving spectral contrasts is manifest as a difference in responses when they move from the excitatory area of the response to the inhibitory and vice versa. The functional significance of the contrast mechanism for the analysis of sound source movement based on direction-dependent spectral features associated with the transfer characteristics of the external ear is discussed.

The selectivity of neurons in the auditory zone of the mouse midbrain to the direction of movement of a spectral notch in wide-band noise.

A series of noise signals was synthesized with spectral notches whose central frequencies moved regularly along the frequency range (from low frequencies to high and from high frequencies to low), imitating movement of the sound source in the vertical plane. The spike responses of neurons in the inferior colliculus of the mouse (Mus musculus) to noise signals changed as the spectral notch moved relative to the excitatory and inhibitory areas of the receptive fields of the neurons and depended on the notch width. Disinhibition reactions in the inhibitory zones were more marked when a frequency notch in the inhibitory zone was followed by a frequency notch in the excitatory part of the response. It is suggested that the selectivity of neurons to the direction of movement of the spectral notch in a noise signal is based on the interaction of the excitatory and inhibitory inputs. The overall set of neuron responses can provide information on the movement of the sound source in acoustic space.

Excitatory and inhibitory intensity tuning in auditory cortex: evidence for multiple inhibitory mechanisms.

The intensity tuning of excitatory and suppressive domain frequency response areas was investigated in 230 cat primary auditory cortical and 92 posterior auditory field neurons. Suppressive domains were explored using simultaneous 2-tone stimulation with one tone at the best excitatory frequency. The intensity tuning of excitatory and suppressive domains was negatively correlated, supporting the hypothesis that inhibitory sidebands are related to excitatory domain intensity tuning. To further test this hypothesis, we compared the slopes of the edges of suppressive bands to the intensity tuning of excitatory domains. Edges of suppressive bands next to excitatory domains had slopes significantly more slanted toward the excitatory area in neurons with intensity-tuned excitatory domains. This relationship was not observed for suppressive band edges not next to the excitatory domain (e.g., the lower edge of lower suppressive bands). This indicates that intensity tuning ultimately observed in the excitatory domain results from overlapping excitatory and inhibitory inputs. In combination with results using forward masking, our results suggest that there are separate early and late sources of inhibition contributing to cortical frequency response areas, and only the early-stage inhibition contributes to excitatory domain intensity tuning.

Interaction of Excitation and Inhibition in Anteroventral Cochlear Nucleus Neurons That Receive Large Endbulb Synaptic Endings

Spherical bushy cells (SBCs) of the anteroventral cochlear nucleus (AVCN) receive their main excitatory input from auditory nerve fibers (ANFs) through large synapses, endbulbs of Held. These cells are also the target of inhibitory inputs whose function is not well understood. The present study examines the role of inhibition in the encoding of low-frequency sounds in the gerbil's AVCN. The presynaptic action potentials of endbulb terminals and postsynaptic action potentials of SBCs were monitored simultaneously in extracellular single-unit recordings in vivo. An input-output analysis of presynaptic and postsynaptic activity was performed for both spontaneous and acoustically driven activity. Two-tone stimulation and neuropharmacological experiments allowed the effects of neuronal inhibition and cochlear suppression on SBC activity to be distinguished. Ninety-one percent of SBCs showed significant neuronal inhibition. Inhibitory sidebands enclosed the high- or low-frequency, or both, sides of the excitatory areas of these units; this was reflected as a presynaptic to postsynaptic increase in frequency selectivity of up to one octave. Inhibition also affected the level-dependent responses at the characteristic frequency. Although in all units the presynaptic recordings showed monotonic rate-level functions, this was the case in only half of the postsynaptic recordings. In the other half of SBCs, postsynaptic inhibitory areas overlapped the excitatory areas, resulting in nonmonotonic rate-level functions. The results demonstrate that the sound-evoked spike activity of SBCs reflects the integration of acoustically driven excitatory and inhibitory input. The inhibition specifically affects the processing of the spectral, temporal, and intensity cues of acoustic signals.

Frequency response areas of neurons in the mouse inferior colliculus. I. Threshold and tuning characteristics.

Excitatory and inhibitory frequency response areas of 130 neurons of the central nucleus of the mouse inferior colliculus (ICC) were mapped by extracellular single-unit recordings and quantitatively evaluated with regard to thresholds, steepness of slopes of excitatory tuning, characteristic frequencies of excitation (CF(E)), inhibition (CFI), and bandwidths of response areas (sharpness of tuning). Two-tone stimuli were used to determine the shapes of inhibitory response areas. Class I neurons (n=54) had asymmetrical (with regard to the CF(E)) excitatory and inhibitory response areas, with inhibition above CF(E) having lower thresholds and covering larger areas than inhibition below CF(E). Quantitative relationships between CF(E) and CF(I) thresholds, and sharpness of tuning showed that the receptive fields of about two-thirds of these neurons had properties similar to auditory nerve fibers. Class II neurons (n=36) had small symmetrical or tilted excitatory areas of rather constant bandwidths and broad inhibitory areas reaching far into and often through the excitatory area, leading to closed excitatory areas in ten neurons. Class III neurons (n=32) had higher excitatory thresholds and the highest proportions of unilateral inhibitory areas compared with neurons of the other classes. Their excitatory area often widened symmetrically with increasing sound level. Their inhibitory areas did not overlap with the excitatory area. Class IV neurons (n=8) had two branches of excitatory areas (two-CFs(E)) and six of the neurons had a central inhibitory area in addition to the low- and high-frequency inhibitory areas. In most neurons, the shapes of excitatory response areas predicted the shapes of inhibitory areas. Altogether, 15 neurons from all 4 classes had areas of facilitation in addition to inhibitory areas. Facilitation in six class IV neurons occurred between the two branches of the excitatory area. All 130 neurons had large inhibitory areas, 106 of them on both sides of the excitatory area. That is, sound processing in the ICC shows strong inhibitory components. The close relationships between excitatory and inhibitory CFs found here indicate that inhibitory projections to and interactions within the ICC are tonotopically organized comparable to the excitatory ones.

Rate representation of tones in noise in the inferior colliculus of decerebrate cats.

Neurons in the central nucleus of the inferior colliculus (ICC) of decerebrate cats show three major response patterns when tones of different frequencies and sound-pressure levels (SPLs) are presented to the contralateral ear. The frequency response maps of type I units are uniquely defined by a narrow excitatory area at best frequency (BF: a unit's most sensitive frequency) and surrounding inhibition at higher and lower frequencies. As a result of this receptive field organization, type I units exhibit strong excitatory responses to BF tones but respond only weakly to broadband noise (BBN). These response characteristics predict that type I units are well suited to encode narrowband signals in the presence of background noise. To test this hypothesis, the dynamic range properties of ICC unit types were measured under quiet conditions and in multiple levels of continuous noise. As observed in previous studies of the auditory nerve and cochlear nucleus, type I units showed upward threshold shifts and discharge rate compression in background noise that partially degraded the dynamic range properties of neural representations at high noise levels. Although the other two unit types in the ICC showed similar trends in threshold shift and noise compression, their ability to encode auditory signals was compromised more severely in increasing noise levels. When binaural masking effects were simulated, only type I units showed an enhanced representation of spatially separated signals and maskers that was consistent with human perceptual performance in independent psychoacoustic observations. These results support the interpretation that type I units play an important role in the auditory processing of narrowband signals in background noise and suggest a physiological basis for spatial factors that govern signal detection under free-field listening conditions.

Single-unit responses in the inferior colliculus of decerebrate cats. I. Classification based on frequency response maps.

This study proposes a classification system for neurons in the central nucleus of the inferior colliculus (ICC) that is based on excitation and inhibition patterns of single-unit responses in decerebrate cats. The decerebrate preparation allowed extensive characterization of physiological response types without the confounding effects of anesthesia. The tone-driven discharge rates of individual units were measured across a range of frequencies and levels to map excitatory and inhibitory response areas for contralateral monaural stimulation. The resulting frequency response maps can be grouped into the following three populations: type V maps exhibit a wide V-shaped excitatory area and no inhibition; type I maps show a more restricted I-shaped region of excitation that is flanked by inhibition at lower and higher frequencies; and type O maps display an O-shaped island of excitation at low stimulus levels that is bounded by inhibition at higher levels. Units that produce a type V map typically have a low best frequency (BF: the most sensitive frequency), a low rate of spontaneous activity, and monotonic rate-level functions for both BF tones and broadband noise. Type I and type O units have BFs that span the cat's range of audible frequencies and high rates of spontaneous activity. Like type V units, type I units are excited by BF tones and noise at all levels, but their rate-level functions may become nonmonotonic at high levels. Type O units are inhibited by BF tones and noise at high levels. The existence of distinct response types is consistent with a conceptual model in which the unit types receive dominant inputs from different sources and shows that these functionally segregated pathways are specialized to play complementary roles in the processing of auditory information.

Lateral inhibition in frequency tuning of central auditory neurons

Previous studies have shown that frequency tuning curves (FTCs) of auditory neurons in the inferior colliculus (IC) are composed of an excitatory area that is either neighbored by an inhibitory area on one flank or is sandwiched by two inhibitory areas. Neurons that are responsible for excitatory and inhibitory areas are likely neighboring neurons. Whereas this neural lateral inhibition sharpens frequency tuning and provides a means to reduce ambiguity in encoding frequency at high stimulus intensities, neurons with two-flank lateral inhibitory areas have larger Qn values and smaller excitatory areas than neurons with one-flank inhibitory areas. The closer the inhibitory area is to the excitatory area, the sharper the frequency tuning becomes. The Qn values of IC neurons tend to increase with the ratio of inhibitory area to excitatory area. Application of bicuculline and/or strychnine broadens FTCs of many IC neurons and completely or partially abolishes the inhibitory areas. Corticofugal pathways sharpen the FTC by narrowing the excitotory area and broadening the lateral inhibitory areas. [Work supported by NSF.]

Sharpening of Frequency Tuning by Inhibition in the Thalamic Auditory Nucleus of the Mustached Bat

Unlike the quasitriangular frequency-tuning curves of peripheral neurons, pencil- or spindle-shaped frequency-tuning curves (excitatory areas) have been found in the central auditory systems of many species of animals belonging to different classes. Inhibitory tuning curves (areas) are commonly found on both sides of such "level-tolerant" sharp frequency-tuning curves. However, it has not yet been examined whether sharpening of frequency tuning takes place in the medial geniculate body (MGB). We injected an inhibitory transmitter antagonist, bicuculline methiodide (BMI), into the MGB of the mustached bat to examine whether frequency tuning is sharpened by inhibition in the MGB and whether this sharpening, if any, occurs in addition to that performed in prethalamic auditory nuclei. Thirty-seven percent of thalamic Doppler-shifted constant frequency (DSCF) neurons mostly showing a level-tolerant frequency-tuning curve had an inhibitory area or areas. BMI changed the inhibitory areas of these neurons into excitatory areas, so that their excitatory frequency-tuning curves became broader. However, the BMI-broadened excitatory frequency-tuning curves were still much narrower than those of peripheral neurons. Our results indicate that level-tolerant frequency tuning of thalamic DSCF neurons is mostly created by prethalamic auditory nuclei and that it is further sharpened in 37% of thalamic DSCF neurons by lateral inhibition occurring in the MGB. The comparisons in sharpness (quality factors) of frequency-tuning curves between peripheral, thalamic, and cortical DSCF neurons indicate that the skirt portion of tuning curves is sharper in the above order, and that their tip portion is not significantly different between the peripheral and thalamic DSCF neurons, but significantly sharper in the cortical DSCF neurons than in the thalamic DSCF neurons. Therefore the central auditory system has inhibitory mechanisms for the progressive sharpening of frequency tuning. DSCF neurons in the primary auditory cortex were recently found to show facilitative responses to paired sounds. That is, they are combination sensitive. In the present studies, we found that thalamic DSCF neurons also showed facilitative responses to paired sounds. The responses of thalamic DSCF neurons to acoustic stimuli consisted of a slow and a fast component. BMI mainly increased the slow component and an excitatory transmitter antagonist, D-2-amino-5-phosphonovalerate mainly suppressed the slow component. Therefore the response pattern of these thalamic neurons is shaped by both gamma-aminobutyric acid-mediated inhibition and N-methyl-D-aspartate-mediated facilitation.