Disproportionate tonotopic representation for processing CF-FM sonar signals in the mustache bat auditory cortex.

The extent of cortical representation of the peripheral sensory field depends on its importance for species behavior. The orientation sound of the mustache bat (Pteronotus parnellii rubiginosus) invariably consists of long constant-frequency and short frequency-modulated components and is indispensable for its survival. A disproportionately large part of the auditory cortex of this bat is occupied by neurons processing the predominant components in the orientation signal and Doppler-shifted echoes. This disproportionate cortical representation related to features of biologically significant signals is comparable to that in the somatosensory and visual systems in many mammals, but it has not previously been observed in the auditory system.

Coordinated activities of middle-ear and laryngeal muscles in echolocating bats.

The middle-ear muscles and laryngeal muscles of the little brown bat (Myotis lucifugus) are highly developed. When the bat emits orientation sounds, action potentials of middle-ear muscles appear approximately 3 milliseconds after those of the laryngeal muscles; this activity of middle-ear muscles attenuates the vocal self-stimulation and improves the performance of the echolocation system. When an acoustic stimulus is delivered, both types of muscles contract; action potentials of the laryngeal muscles appear approximately 3 milliseconds after those of the middle-ear muscles. These two groups of muscles are apparently activated in a coordinated manner not only by the nerve impulses from the vocalization center, but also by those from the auditory system.

Echo frequency selectivity of duration-tuned inferior collicular neurons of the big brown bat, Eptesicus fuscus, determined with pulse-echo pairs.

During hunting, insectivorous bats such as Eptesicus fuscus progressively vary the repetition rate, duration, frequency and amplitude of emitted pulses such that analysis of an echo parameter by bats would be inevitably affected by other co-varying echo parameters. The present study is to determine the variation of echo frequency selectivity of duration-tuned inferior collicular neurons during different phases of hunting using pulse-echo (P-E) pairs as stimuli. All collicular neurons discharge maximally to a tone at a particular frequency which is defined as the best frequency (BF). Most collicular neurons also discharge maximally to a BF pulse at a particular duration which is defined as the best duration (BD). A family of echo iso-level frequency tuning curves (iso-level FTC) of these duration-tuned collicular neurons is measured with the number of impulses in response to the echo pulse at selected frequencies when the P-E pairs are presented at varied P-E duration and gap. Our data show that these duration-tuned collicular neurons have narrower echo iso-level FTC when measured with BD than with non-BD echo pulses. Also, IC neurons with low BF and short BD have narrower echo iso-level FTC than IC neurons with high BF and long BD have. The bandwidth of echo iso-level FTC significantly decreases with shortening of P-E duration and P-E gap. These data suggest that duration-tuned collicular neurons not only can facilitate bat's echo recognition but also can enhance echo frequency selectivity for prey feature analysis throughout a target approaching sequence during hunting. These data also support previous behavior studies showing that bats prepare their auditory system to analyze expected returning echoes within a time window to extract target features after pulse emission.

Electrical stimulation of bat superior colliculus influences responses of inferior collicular neurons to acoustic stimuli.

The influence of electrical stimulation of the superior colliculus (SC) on acoustically evoked responses of inferior collicular (IC) neurons was examined in 24 barbiturate-anesthetized Rufous horseshoe bats, Rhinolophus rouxi. Acoustic stimuli (50 ms, 0.5 ms rise-decay times) were delivered from a loudspeaker placed 68 cm in front of each bat and a total of 354 IC neurons were isolated. The response latencies of these neurons were mainly between 7.5 and 17.5 ms. When the ipsilateral SC was electrically stimulated, responses of 227 (64%) neurons were not affected, but responses of the remaining (127 neurons, 36%) were either inhibited (102 neurons, 29%) or facilitated (25 neurons, 7%). The degree of inhibition and the response latency of the inhibited neurons increased with the amplitude of electrical stimulation. Inhibition of a neuron's activity was also dependent upon the time interval between acoustic and electrical stimuli. The best inhibitory latency measured at maximal inhibition was between 12 and 20 ms. Conversely, facilitation shortened the response latency of IC neurons and the degree of facilitation increased with the amplitude of the acoustic stimulus. Since the SC plays an essential role in orienting an animal's responses toward sensory stimuli, our findings suggest that the SC may affect the processing of acoustic signals in the auditory system during acoustically guided orientation.

The role of GABAergic inhibition on direction-dependent sharpening of frequency tuning in bat inferior collicular neurons.

This study examined the role of GABAergic inhibition on direction-dependent sharpening of frequency tuning curves (FTCs) in bat inferior collicular (IC) neurons under free field stimulation conditions. The minimum threshold (MT) at the neurons best frequency (BF) and the sharpness (Q(10), Q(20), Q(30)) of FTCs of most IC neurons increased as the sound direction changed from contralateral azimuths to ipsilateral azimuths. The application of GABA(A) antagonist, bicuculline, lowered all MTs but the application did not abolish direction-dependent variation in MT. MTs determined during bicuculline application at 40 ipsilateral were still significantly higher than those determined at 40 degrees contralateral (two-tailed paired t-test, P<0.0001). In contrast, although application of bicuculline essentially had no effect on the BFs of IC neurons, it differentially broadened neurons FTCs at different azimuths abolishing the direction-dependent sharpening of frequency tuning (i. e. Q(n) values, two-tailed paired t-test, P<0.01). These data indicate that GABAergic inhibition makes an important contribution to the direction-dependent frequency tuning of most IC neurons.

Corticofugal inhibition compresses all types of rate-intensity functions of inferior collicular neurons in the big brown bat.

Recent studies have shown that the auditory corticofugal system modulates and improves signal processing in the frequency, time and spatial domains. In this study, we examine corticofugal modulation of rate-intensity functions of inferior collicular (IC) neurons of the big brown bat, Eptesicus fuscus, by electrical stimulation in the primary auditory cortex (AC). Cortical electrical stimulation compressed all types of rate-intensity functions so as to increase the slope but decrease the dynamic range of IC neurons. Cortical electrical stimulation also shifts the responsive intensity of IC neurons to higher levels. These data indicate that corticofugal modulation also improves subcortical signal processing in intensity domain. The implication of these findings to bat echolocation is discussed.

Neurons in the cerebellum of echolocating bats respond to acoustic signals.

Single neurons responding to auditory stimuli (40 msec duration, 0.5 msec rise-decay time) could be isolated from rather large areas of the cerebellar vermis and hemispheres of an echolocating bat, Eptesicus fuscus. These neurons had latencies between 4 and 13 msec and best frequencies between 22 and 77 kHz. The Q10-dB values of their tuning curves were between 1.4 and 16.6. When acoustic stimuli were delivered though the earphones, tuning curves measured from each ear alone were nearly identical in shape and best frequency. The minimum thresholds of these neurons were between 12 and 65 dB SPL. Apparently, these are suitable for reception of the bat's echolocating signals.

Pinna orientation determines the maximal directional sensitivity of bat auditory neurons.

The auditory response areas of 192 inferior collicular neurons (IC) of Eptesicus fuscus were studied under free field acoustic stimulation. The boundary of the auditory response area of a neuron expands with stimulus intensity (Fig. 1). However, there is a response center within each neuron's response area at which the neuron has the maximal sensitivity. All response centers of the 192 neurons are located within a limited space of the bat's contralateral auditory space. The position of the response center of a neuron changes with different pinna orientations (Figs. 2 and 3) providing a bat with versatility in maximizing the sensitivity of its echolocation system.

corticofugal regulation of excitatory and inhibitory frequency tuning curves of bat inferior collicular neurons.

Corticofugal regulation of excitatory and inhibitory frequency tuning curves (FTCs) of neurons in the central nucleus of bat inferior colliculus (ICc) was studied by electrical stimulation of the primary auditory cortex (AC stimulation) under free field stimulation conditions using a two-tone inhibition paradigm. AC stimulation narrowed the excitatory FTCs and asymmetrically expanded the lateral inhibitory FTCs of corticofugally inhibited ICc neurons. The opposite results were observed for excitatory and inhibitory FTCs of corticofugally facilitated ICc neurons. These data support previous reports that corticofugal systems work together with widespread lateral inhibition to regulate subcortical frequency processing.

Fos-like immunoreactivity elicited by sound stimulation in the auditory neurons of the big brown bat Eptesicus fuscus.

C-fos immunocytochemistry was used as a rapid and sensitive technique for identification of sound activated neurons in the cerebral cortex, the cerebellum and subcortical nuclei of the big brown bat, Eptesicus fuscus. When bats were stimulated with sounds under the both-ears opened conditions, Fos-like immunoreactive neurons were bilaterally and symmetrically distributed in all subcortical auditory nuclei, the auditory cortex, the superior colliculus, the pontine nuclei and the cerebellar deep nuclei. Interestingly, when bats were stimulated with sounds under the monaurally plugged conditions, a larger (31-74% more) number of Fos-like immunoreactive neurons were observed. They were predominantly distributed in all contralateral auditory nuclei from the level of the nucleus of the lateral lemniscus down and in all ipsilateral auditory nuclei from the level of inferior colliculus up as well as in the contralateral superior colliculus, pontine nuclei and cerebellar deep nuclei. Implications of these observations in relation to known mammalian auditory pathways and electrophysiological studies are discussed.

Pulse repetition rate increases the minimum threshold and latency of auditory neurons.

The effect of pulse repetition rate on auditory sensitivity of the big brown bat, Eptesicus fuscus, was studied by determining the minimum threshold, response latency and recovery cycle of inferior collicular neurons at different repetition rates under free field stimulation conditions. In general, collicular neurons shortened the response latency and increased the number of impulses monotonically or non-monotonically with stimulus intensity. They recovered at least 50% when the interpulse interval was 10-57 ms. In addition, they increased the minimum threshold, lengthened the response latency, and reduced the number of impulses discharged to each pulse with increasing repetition rate. The increase in minimum threshold with repetition rate is partly because the neuron can not recover from previous stimulation when the interpulse interval is shortened. This increase reduces a neuron's response sensitivity and thus diminishes its number of impulses to each presented pulse. This increase also reduces the effectiveness of a given stimulus intensity which contributes to the lengthening of the neuron's response latency. Data obtained from single neuron recordings are used to highlight these observations. Implications of present findings regarding the bat's echolocation are also discussed.

Auditory response properties and directional sensitivity of cerebellar neurons of the echolocating bat, Eptesicus fuscus.

Auditory response properties and directional sensitivity of cerebellar neurons of Eptesicus fuscus were studied under free-field stimulation conditions. The best frequency (BF) and minimum threshold (MT) of a recorded neuron were first determined with a sound delivered in front of the bat. Discharge pattern and MT were studied with both BF stimuli and one-octave downward and upward sweep FM (frequency-modulated) stimuli. The directional sensitivity of cerebellar neurons was then studied by determining the variation of MT and response latency with BF and FM stimuli broadcast from each of 15 loudspeakers attached to a semicircular wooden track in front of the bat. All 85 cerebellar neurons recorded discharged phasically to acoustic stimuli. Only 20 were spontaneously active. Cerebellar neurons were generally more sensitive to FM stimuli than to pure tone pulses. Thus, they discharged more vigorously and had a lower MT to the former than the latter stimulus. Directional sensitivity of 47 neurons (BF = 23.4-81.1 kHz) was studied. All neurons varied their MTs with sound direction. Most neurons (n = 37, 79%) showed a lowest MT to a frontal sound. Directional sensitivity of cerebellar neurons appears to be sharper when determined with BF tone pulses than with FM stimuli. Thus the directional slope and the difference in MT between the best and worst angles of these neurons were larger when determined with the BF stimulus. Directional sensitivity of cerebellar neurons is not dependent upon stimulus frequency, unlike that of the inferior and cortical neurons of the same bat. Cerebellar neurons also varied their response latency with sound direction. Such a variation may provide the bat with another neural code for sound localization.

Auditory space representation in the superior colliculus of the big brown bat, Eptesicus fuscus.

The auditory response areas of 123 superior collicular (SC) units of Eptesicus fuscus were studied under free-field acoustic stimulus conditions. A stimulus was delivered from a loudspeaker placed 14 cm in front of a bat. The best frequency of a unit was determined by changing the stimulus frequency until the minimum threshold was measured. A best frequency stimulus was then delivered as the loud-speaker was moved across the auditory space to determine the response center of the auditory response area of each unit. The response center was defined as the direction at which the unit had its lowest minimum threshold. The stimulus intensity was then raised 2-20 dB above the lowest minimum threshold of the unit and the response area for each stimulus intensity was determined. The response area of a unit expands with stimulus intensity, but the expansion is not even in all directions. The size of the response area of a unit does not correlate with its minimum threshold, best frequency, or recording depth. Response centers of 7 units were located directly in front of the animal, but most response centers were located in a limited portion of the contralateral auditory space. Although each unit has a response center which is the point of maximal sensitivity, the point-to-point representation of the auditory space is not systematically organized. We suggest that an animal with highly mobile external pinnae may not need an orderly auditory space map in its neural tissue for accurate sound localization.

Auditory response properties and spatial response areas of single neurons in the pontine nuclei of the big brown bat, Eptesicus fuscus.

Using free-field acoustic stimulation conditions, we studied the response properties and spatial sensitivity of 146 pontine neurons of the big brown bat, Eptesicus fuscus. The best frequency (BF) and minimum threshold (MT) of a pontine neuron were first determined with a sound broadcast from a loudspeaker placed ahead of the bat. A BF sound was delivered from the loudspeaker as it moved across the frontal auditory space in order to locate the response center at which the neuron had its lowest MT. Then the basic response properties of the neuron to a sound delivered from the response center were studied. As in inferior collicular and auditory cortical neurons, pontine neurons can be characterized as phasic responders, phasic bursters and tonic responders. They have both monotonic and non-monotonic intensity-rate functions. However, most of them are broadly tuned as are cerebellar neurons. Auditory spatial sensitivity was studied for 144 pontine neurons. In 9 neurons, variation of MT with a BF sound delivered from several azimuthal and elevational angles along the horizontal and vertical planes crossing the neuron's response center was measured. In addition, variation in the number of impulses with several stimulus intensities at 10 dB increments above a neuron's MT delivered from each angle was also studied. The auditory spatial sensitivity of other pontine neurons was studied by measuring the response area of each neuron with stimulus intensities at 3, 5, 10, 15 or 40 dB above its lowest MT. The response areas of pontine neurons expanded asymmetrically with stimulus intensity, but the size of the response area was not correlated with either MT or BF. In half of the pontine neurons studied, the response area expanded greatly and eventually covered almost the entire frontal auditory space. The response areas of the other half of the pontine neurons only expanded to a restricted area of frontal auditory space. Two possible neural mechanisms underlying these two types of response areas are hypothesized. The response centers of all 144 neurons were located within a small area of the frontal auditory space. The locations of response centers of these neurons are not correlated with their BFs. The distribution pattern of these response centers is comparable to that of superior collicular and cerebellar neurons but is different from that of inferior collicular and auditory cortical neurons. The results of our study suggest that auditory information is integrated in the pontine nuclei before being further sent into the cerebellum.

Responses of cerebellar neurons of the CF-FM bat, Pteronotus parnellii to acoustic stimuli.

Single units (125) which faithfully discharged action potentials to acoustic stimuli (35 ms in duration with 0.5 ms rise and decay times) were recorded in the cerebellar vermis and hemispheres of the CF-FM bat, Pteronotus parnellii. These units had response latencies between 1.5 and 27 ms and minimum thresholds between 2 and 83.5 dB SPL. Best frequencies (BFs) of these units ranged from 30.32 to 79.28 kHz, but more than half (64 units, 51.2%) were between 59.73 and 63.32 kHz. While most tuning curves of these units were either broad or irregular, those curves with BFs tuned at around 61 kHz which is the frequency of the predominant CF component of the bat's echolocation signals were extremely narrow with Q10-dB values as high as 153. Those units (29) with BFs tuned near the 61 kHz also showed off-responses. These data indicate that auditory specialization for processing of species-specific orientation signals also exists in the cerebellum of this bat.

Mapping of the auditory area in the cerebellar vermis and hemispheres of the mustache bat, Pteronotus parnellii parnellii.

Microelectrode mapping of the auditory areas in the cerebellar vermis and hemispheres of mustache bats, Pteronotus parnellii parnellii, reveals that a large area of the bat's cerebellum contains units responding to acoustic signals. A study of frequency tuning of isolated units shows that there are two large groups of auditory units. The units of one group are sharply tuned to a very narrow band of frequency with BFs between 60 and 64 kHz. The units of the other group are broadly tuned, with BFs between 47 and 59 kHz. These two groups of units are probably involved in processing the predominant CF and FM portions of the bat's orientation sounds during echolocation.

Auditory spatial sensitivity of inferior collicular neurons of echolocating bats.

The sensitivity of 94 inferior collicular (IC) neurons of Eptesicus fuscus and Myotis lucifugus to spatial location of the acoustic stimulus were studied under free-field stimulus conditions. The best frequency (BF) and minimum threshold (MT) of each neuron were determined with sound delivered in front of the bat. Then the variation in discharge rate of the neuron was measured with a BF sound broadcast from a moving loudspeaker at different angular positions along the horizontal, vertical or diagonal plane of the frontal auditory space. A wide range of stimulus intensities above the MT of the neuron was used. Neurons were classified into 3 classes on the basis of their spatial sensitivity: (1) omnisensitive neurons (15%) were broadly tuned to sound delivered in the frontal auditory space and their responses did not show any correlation with sound location; (2) stimulus intensity-dependent neurons (28%) varied their discharge rates with sound location and intensity so that the peak of their spatial response profiles also varied with stimulus intensity; and (3) stimulus intensity-independent neurons (57%) varied their discharge rates only with sound location over a wide range of stimulus intensities so that their peak discharge always appeared at the same or a small range of angle. In most cases, the medial limbs of the spatial sensitivity curve for these neurons were extremely sharp and congruent. By moving the loudspeaker along the horizontal, vertical and diagonal planes, it was possible to approximate the boundary of the spatial response area of a neuron. Most IC neurons responded to sound delivered within 20 degrees ipsilateral, 60 degrees contralateral, 45 degrees up and 40 degrees down of the frontal auditory space, confirming previous similar studies. In general, an increasing stimulus repetition rate appeared to sharpen the spatial sensitivity curve of a neuron. Conversely, an increasing moving velocity of the stimulus decreased its response. The possible role of these 3 classes of neurons in echolocation and neural mechanisms underlying the spatial sensitivity of these neurons is discussed.

Neurons in the superior colliculus of echo-locating bats respond to ultrasonic signals.

Using conventional electrophysiological techniques, we demonstrate that neurons in the superior colliculus of the big brown bat (Eptesicus fuscus) respond to ultrasonic signals. Most response properties of these neurons are very similar to neurons of the inferior colliculus in the same bat.

Neurons in the inferior colliculus, auditory cortex and pontine nuclei of the FM bat, Eptesicus fucus respond to pulse repetition rate differently.

Single-neuron responses to pulse repetition rate in the inferior colliculus, auditory cortex and pontine nuclei of the FM bat, Eptesicus fuscus were studied under free-field stimulation conditions. The best frequency (BF) and minimum threshold (MT) of each neuron were first determined with a 4 ms pulse broadcast from a specific point (response center) of the bat's frontal auditory space at which the neuron had maximal spatial sensitivity. The neuron's intensity-rate function was then studied with a 4 ms BF pulse delivered at 10 dB increments above its MT in order to determine the best intensity to which the neuron discharged maximally. The neuron's discharge pattern and number of impulses to 32 trials of 300 ms train stimuli, which consisted of different number of 4 ms BF and best intensity pulses (1, 2, 3, 8, 12, 19, 24, 29 pulses/train) and delivered at an interpulse interval of 1000, 250, 170, 100, 40, 25, 15, 12 and 10 ms (i.e. at a pulse repetition rate of 1, 4, 6, 10, 25, 40, 67, 83, 100 pulses/s), were sequentially recorded. All neurons recorded from the inferior colliculus, auditory cortex and pontine nuclei discharged phasically (1-3 impulses) but they responded to the pulse repetition rate in different manners. More than 63% of 38 inferior collicular and 65 pontine neurons studied discharged impulses to each pulse within a train stimulus when the pulse repetition rate was up to 40 pulses/s.(ABSTRACT TRUNCATED AT 250 WORDS)

Tracing the auditory pathways to electrophysiologically characterized neurons with HRP and Fos double-labeling technique.

By combining HRP histochemistry with Fos immunocytochemistry, we demonstrate in this study that electrophysiologically characterized auditory neurons can be double-labeled with HRP and Fos after iontophoretic injection of HRP into the recording site. Neurons which projected fibers to the recording site were labeled with HRP and were Fos-like immunoreactive. This double-labeling technique in combination with electrophysiological recording offers the possibility to determine the fiber projections between sound-activated neurons which are identified either electrophysiologically and/or immunocytochemically.