Junsei Horikawa

Optical imaging of spatiotemporal patterns of glutamatergic excitation and GABAergic inhibition in the guinea-pig auditory cortex in vivo.

1. Glutamatergic excitation and gamma‐aminobutyric acid (GABA)‐ergic inhibition in layers II and III of the auditory cortex of anaesthetized guinea‐pigs were recorded optically using a voltage‐sensitive dye RH795 and a 12 x 12 photodiode array. 2. After contralateral ear stimulation with pure tones, transient excitatory responses followed by inhibitory responses were observed in fields A (primary) and DC of the auditory cortex. The area of the excitatory responses was sandwiched or surrounded by the areas of the inhibitory responses. 3. Optically recorded excitatory responses to pure tones had two components: a component sensitive to 6‐cyano‐7‐nitroquinoxaline‐2,3‐dione (CNQX), a non‐N‐methyl‐D‐aspartate (non‐NMDA) receptor antagonist, and a component sensitive to 2‐amino‐5‐phosphono‐valerate (APV), an NMDA receptor antagonist. Application of CNQX (5 microM) to the auditory cortex suppressed an early, but not a late, phase of the excitation; application of APV (100 microM) had the opposite effect. Concomitant application of CNQX and bicuculline methiodide (BMI, 4 microM), a GABAA receptor antagonist, increased the amplitude of the late phase 4‐fold. This enhanced response was suppressed by APV. 4. These results indicate that (i) auditory cortical excitatory responses are mediated by both non‐NMDA and NMDA receptors, (ii) inhibition is mediated by GABAA receptors, (ii) the excitatory bands are sandwiched or surrounded by GABAA receptor‐mediated inhibitory areas and (iv) GABAA receptors effectively inhibit the NMDA, but not the non‐NMDA, receptor‐mediated excitation.

The columnar and layer-specific response properties of neurons in the primary auditory cortex of Mongolian gerbils

The columnar and layer-specific response properties of neurons in the primary auditory cortex (AI) of Mongolian gerbils were studied using single-unit recordings of responses to tone-burst stimuli presented to the ear contralateral to the recording side. During near-radial microelectrode penetrations of the AI in 100-microm steps, the best frequency (BF), best threshold (BT), best amplitude (BA), latency, tuning curve and Q10dB were recorded. Neurons encountered during single penetrations showed similar BFs, indicating a columnar frequency organization, but their latencies and Q10dBs differed. The BAs and BTs recorded within single penetrations often showed a similar value in the middle cortical layers. The latencies and Q10dBs of these neurons exhibited a tendency toward a layer-specific distribution. The latencies of neurons located in layers I-V were longer than those located in layer VI. The Q10dBs of neurons located in layers III and IV were higher than those located in layers I and VI. These results are almost consistent with those of previous studies on frequency representation, and indicated the existence of an integrative mechanism of frequency processing in the AI. This is the first study in which a layer-specific, partially columnar organization for stimulus amplitude is described.

Spatio-temporal pattern of frequency representation in the auditory cortex of guinea pigs

The spatio-temporal pattern of sound-evoked neural activity in the guinea pig auditory cortex was studied by optical recording with the aid of voltage-sensitive dye. Changes in light intensity induced by sounds at various frequencies and pressure levels were recorded with a 12 x 12 array of photodiodes. The amplitudes of the responses were displayed as sequential two-dimensional images. Tonotopical organization was found in two subdivisions of the auditory cortex, the anterior field (field A) and the dorsocaudal field (field DC). The frequency gradients in fields A and DC had a mirror-image relationship. This agrees with results obtained by the microelectrode technique. However, the tonotopic response observed in our study was transient. The focal activity that began in field A propagated in two directions; dorsally along the iso-frequency bands in field A, and caudally toward field DC. This suggests that the sound information processing initiates at field A, and its outputs are transferred to field DC, which is probably a hierarchically higher center.

The acoustic middle ear muscle reflex in albino rats

The acoustic middle ear muscle reflex was studied in albino rats anesthetized with chloralose. The best frequency of the reflex and the threshold at this frequency were on average about 3 kHz and 57 dB SPL, respectively. The threshold increased as frequency increased above, and decreased below, the best frequency at a rate of about 20 dB/octave. Above about 12 kHz, the muscular response showed instability and habituation. Thresholds were similar between stapedius and tensor tympani reflexes and between ipsilateral and contralateral reflexes. The middle ear transmission loss due to the reflex was the greatest and nearly constant below about 1 kHz, where the loss was about 18 dB at the maximal stimulation. Above this frequency the loss decreased as frequency increased up to 20 kHz. Thus the reflex, unlike that in other animals, suppressed transmission over the whole range of reflex-eliciting frequencies. The transfer function of the reflex had a well damped low-pass characteristic with a cut-off frequency of about 20 Hz. From the above characteristics of the reflex, the role of the rats tympanic muscles in improving ultrasonic hearing under ambient noises was suggested.

Optical imaging of dynamic horizontal spread of excitation in rat auditory cortex slices.

Optical recordings using a voltage-sensitive dye (RH482) were conducted in brain slice preparations to investigate spatiotemporal patterns of excitation in the rat auditory cortex. Electrical stimulation of the border between the white matter and layer VI evoked vertical as well as horizontal spreading responses. While velocities of vertical and horizontal propagation of excitation were similar to those reported in non-disinhibited preparations, the horizontal propagation was widespread and strong especially in layers II/III in auditory cortex slices. This horizontal spread was blocked by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) but not by D-2-amino-5-phosphonopentanoic acid (D-AP5). These results suggest that the horizontal responses, especially in layers II/III, are prominent and are mediated primarily by non-N-methyl-D-aspartic acid (NMDA) receptors in the auditory cortex.

Mechanical and neural interactions between binaurally applied sounds in cat cochlear nerve fibers

Most single fibers of the cochlear nerve (CN) in 22 cats exhibited effects of mechanical interaction in one cochlea between two sounds applied binaurally, similarly to results in two cats in which the contralateral CN was transected. In 11 of 189 fibers, the spontaneous and/or the sound-evoked activity was suppressed by a contralateral intense best-frequency sound; this indicates an interaural neural inhibition, probably through the olivocochlear bundle (OCB). The inhibited fiber population was small, and the intensity differences between the binaural sounds were exceptionally large, so that a simple negative feed-back function via the OCB is not likely.

Optical Imaging of Neural Activity in Auditory Cortex Induced by Intracochlear Electrical Stimulation

Little is known about the representation of electrically evoked activity in the auditory cortex. We observed evoked activity in guinea pig auditory cortex evoked by acoustical and electrical stimulation to the cochlea by optical imaging with the aid of a voltage-sensitive dye. Light signals from the cortex were recorded with a 12 x 12 array of photodiodes, and transferred to the spatio-temporal images by every 0.57 ms. The activity by pure tones was shown spatio-temporally through tonotopical organization in the cortex according to the sound frequencies. The tonotopic responses were dynamically changed. When the cochlea was stimulated with single electrical pulses, focal activities were observed in the cortex as spatio-temporal patterns. Activated cortical regions were not sharply localized, but varied with stimulating positions of the cochlea. The curves of response magnitude versus stimulus intensity showed the narrow dynamic range, and that of latency was almost constant. These results were significantly different from those for normal sound stimulation.

Optical study of spatiotemporal inhibition evoked by two-tone sequences in the guinea pig auditory cortex

Abstract Spatiotemporal response patterns in the anterior and dorsocaudal fields of the guinea pig auditory cortex after two-tone sequences were studied in anesthetized animals (Nembutal 30 mg kg−1) using an optical recording method (voltage-sensitive dye RH795, 12 × 12 photodiode array). Each first (masker) and second (probe) tone was 30 ms long with a 10-ms rise-fall time. Masker-probe pair combinations of the same or different frequencies with probe delays of 30–150 ms were presented to the ear contralateral to the recording side. With same-frequency pairs, responses to the probe were inhibited completely after probe delays of less than 50 ms and the inhibition lasted for more than 150 ms, and the inhibition magnitudes in different isofrequency bands of the anterior field were essentially the same. With different-frequency (octave-separated) pairs, responses to the probe were not inhibited completely even after probe delays as short as 30 ms, and the inhibition lasted only for 110–130 ms. Inhibition magnitudes were different from location to location.

REAL-TIME IMAGING OF NEURAL ACTIVITY DURING BINAURAL INTERACTION IN THE GUINEA PIG AUDITORY CORTEX

Abstract Spatio-temporal patterns of binaural interaction in the guinea pig auditory cortex (AC) were observed using optical recording with a 12 × 12 photodiode array and a voltage-sensitive dye. The amplitudes of the sound-induced light signals from the cortex were transformed into sequential two-dimensional images every 0.58 ms. Binaural sound stimuli evoked an excitatory response followed by a strong inhibition, and contralateral stimuli evoked a strong excitatory response followed by a weak inhibition. Ipsilateral sound stimuli evoked a weak response. Binaural stimulation induced two types of ipsilateral inhibition: a fast binaural inhibition which was detected only after the contralateral and ipsilateral responses were subtracted from the binaural responses, and which appeared 12–25 ms after the onset of stimulation, and a slow binaural inhibitory effect which was clearly observed in the binaural responses themselves, appearing 70–95 ms after the onset of stimulation. The fast binaural inhibition was observed in the same area as the contralateral excitatory response. The inhibited area became stronger and more widespread with increasing intensity of ipsilateral stimulation. We did not observe the specialized organization of binaural neurons as electrophysiologically found in the cat AC, in which binaural neurons of the same binaural response type are clustered together and alternate with clusters of other response types.

Spatiotemporal representation of binaural difference in time and intensity of sound in the guinea pig auditory cortex.

Neural activity of the auditory cortex (AC) in response to a change of interaural intensity difference (IID) and interaural time difference (ITD) of sound stimuli was observed by optical recording with a 12 x 12 photodiode array and the voltage-sensitive dye, RH795. Guinea pigs (280-450 g) were anesthetized with sodium pentobarbital (30 mg/kg) and supplemental doses of neuroleptic solutions. When both ears were stimulated dichotically by tone bursts (14 kHz, 75 dB SPL), excitatory optical signals appeared in both anterior (A) and dorsocaudal (DC) fields of AC. An increase of intensity of ipsilateral stimulation from 65 to 95 dB SPL caused a decrease of neural activity of isofrequency bands in both fields. An increase of ipsilateral leads from -2.5 to 10 ms resulted in a gradual decrease of the amplitude of the excitatory responses. A strong inhibition was observed in field DC and the ventral portion of field A. These results show the different spatiotemporal representation of IID and ITD sensitivities in AC. However, the ipsilateral lead inducing a large inhibition was much longer than the time difference (80 micros) calculated from the interaural distance of the guinea pig. This indicates that the longer binaural inhibition observed in AC would have a different functional significance from that of the neural system of ITD detection in the guinea pig.