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Katsuei Shibuki

Publications and source records attributed to Katsuei Shibuki.

At least 19 recordsLinked to original sources

Requirement of the auditory association cortex for discrimination of vowel-like sounds in rats.

We investigated the roles of the auditory cortex in discrimination learning of vowel-like sounds consisting of multiple formants. Rats were trained to discriminate between synthetic sounds with four formants. Bilateral electrolytic lesions including the primary auditory cortex and the dorsal auditory association cortex impaired multiformant discrimination, whereas they did not significantly affect discrimination between sounds with a single formant or between pure tones. Local lesions restricted to the dorsal/rostral auditory association cortex were sufficient to attenuate multiformant discrimination learning, and lesions restricted to the primary auditory cortex had no significant effects. These findings indicate that the dorsal/rostral auditory association cortex but not the primary auditory cortex is required for discrimination learning of vowel-like sounds with multiple formants in rats.

Acoustic Stimulation↗

Functional local connections with differential activity-dependence and critical periods surrounding the primary auditory cortex in rat cerebral slices.

Sensory information is processed in neural networks connecting the primary sensory cortices with surrounding higher areas. Here, we investigated the properties of local connections between the primary auditory cortex (area 41) and surrounding areas (areas 20, 36, 18a and 39) in rat cerebral slices. Neural activities elicited by repetitive electrical stimulation were visualized using the activity-dependent changes in endogenous fluorescence derived from mitochondrial flavoproteins, which mostly reflect activities produced by polysynaptic glutamatergic transmission. Polysynaptic feedforward propagation was dominant compared with the corresponding polysynaptic feedback propagation between the primary (area 41) and secondary (areas 20 and 36) auditory cortices, while such a tendency was less clear in other pathways. Long inter-areal (>1 mm) propagation with the same dominancy was observed after layer V stimulation between areas 41 and 20, and was not affected by cutting the underlying white matter. Activity-dependent changes in neural activities induced by low-frequency stimulation in the presence of 1 microM bicuculline were investigated using Ca2+ imaging. Significant potentiation of the polysynaptic Ca2+ activities was only observed in polysynaptic feedforward pathways from the primary to secondary auditory cortices. Experience-dependence of the connections between areas 41 and 20 was investigated using flavoprotein fluorescence imaging. The activities from areas 41 to 20 were reduced by cochlear lesions produced at P12 but not at P28, while the activities from areas 20 to 41 were reduced by the lesions at P28, suggesting the critical period for the polysynaptic feedforward connection was before P28, while for the polysynaptic feedback connection was after P28.

Animals↗

Enduring critical period plasticity visualized by transcranial flavoprotein imaging in mouse primary visual cortex.

Experience-dependent plasticity in the visual cortex was investigated using transcranial flavoprotein fluorescence imaging in mice anesthetized with urethane. On- and off-responses in the primary visual cortex were elicited by visual stimuli. Fluorescence responses and field potentials elicited by grating patterns decreased similarly as contrasts of visual stimuli were reduced. Fluorescence responses also decreased as spatial frequency of grating stimuli increased. Compared with intrinsic signal imaging in the same mice, fluorescence imaging showed faster responses with approximately 10 times larger signal changes. Retinotopic maps in the primary visual cortex and area LM were constructed using fluorescence imaging. After monocular deprivation (MD) of 4 d starting from postnatal day 28 (P28), deprived eye responses were suppressed compared with nondeprived eye responses in the binocular zone but not in the monocular zone. Imaging faithfully recapitulated a critical period for plasticity with maximal effects of MD observed around P28 and not in adulthood even under urethane anesthesia. Visual responses were compared before and after MD in the same mice, in which the skull was covered with clear acrylic dental resin. Deprived eye responses decreased after MD, whereas nondeprived eye responses increased. Effects of MD during a critical period were tested 2 weeks after reopening of the deprived eye. Significant ocular dominance plasticity was observed in responses elicited by moving grating patterns, but no long-lasting effect was found in visual responses elicited by light-emitting diode light stimuli. The present results indicate that transcranial flavoprotein fluorescence imaging is a powerful tool for investigating experience-dependent plasticity in the mouse visual cortex.

Animals↗

Transcranial imaging of audiogenic epileptic foci in the cortex of DBA/2J mice.

Exposure to intense sound stimuli induces audiogenic seizures in DBA/2J mice. We investigated cortical activities during sound stimulation using flavoprotein fluorescence imaging. Most DBA/2J mice had seizures during intense sound stimulation, with more than half surviving after seizures. Surviving mice were anesthetized with urethane (1.6 g/kg, intraperitoneal), and the skull was exposed and then covered with clear resin. More than 3 days after surgery, the mice were lightly anesthetized with urethane (0.8 g/kg) and cortical activities during intense sound stimulation were visualized. Focal responses appeared near the somatosensory cortex together with spike activities localized in the response area. These findings indicate that epileptic foci of audiogenic seizure are formed in the cortex of DBA/2J mice.

Acoustic Stimulation↗

Transcranial fluorescence imaging of auditory cortical plasticity regulated by acoustic environments in mice.

Functional brain imaging using endogenous fluorescence of mitochondrial flavoprotein is useful for investigating mouse cortical activities via the intact skull, which is thin and sufficiently transparent in mice. We applied this method to investigate auditory cortical plasticity regulated by acoustic environments. Normal mice of the C57BL/6 strain, reared in various acoustic environments for at least 4 weeks after birth, were anaesthetized with urethane (1.7 g/kg, i.p.). Auditory cortical images of endogenous green fluorescence in blue light were recorded by a cooled CCD camera via the intact skull. Cortical responses elicited by tonal stimuli (5, 10 and 20 kHz) exhibited mirror-symmetrical tonotopic maps in the primary auditory cortex (AI) and anterior auditory field (AAF). Depression of auditory cortical responses regarding response duration was observed in sound-deprived mice compared with naïve mice reared in a normal acoustic environment. When mice were exposed to an environmental tonal stimulus at 10 kHz for more than 4 weeks after birth, the cortical responses were potentiated in a frequency-specific manner in respect to peak amplitude of the responses in AI, but not for the size of the responsive areas. Changes in AAF were less clear than those in AI. To determine the modified synapses by acoustic environments, neural responses in cortical slices were investigated with endogenous fluorescence imaging. The vertical thickness of responsive areas after supragranular electrical stimulation was significantly reduced in the slices obtained from sound-deprived mice. These results suggest that acoustic environments regulate the development of vertical intracortical circuits in the mouse auditory cortex.

Acoustic Stimulation↗

Roles of the auditory cortex in discrimination learning by rats.

We investigated the roles of the auditory cortex in sound discrimination learning in Wistar rats. Absolute pitch or relative pitch can be used as discrimination cues in sound frequency discrimination. To clarify this, rats were trained to discriminate between rewarded (S+) and unrewarded (S-) test stimuli (S+ frequency>S- frequency). After learning was acquired by rats, performance was tested in a new test in which S+ frequency was constant but S+ frequency S- frequency but both frequencies were increased. If the discrimination cue of the first test was preserved in the new test, performance following change of testing procedures was expected to remain high. The measured performance suggested that rats used relative pitch in half octave discrimination (difference between S+ and S- frequencies, 0.5 octave), and absolute pitch in octave discrimination (difference between S+ and S- frequencies, 1.0 octave). Bilateral lesions in the auditory cortex had almost no effect on performance before procedure change. Furthermore, performance following procedure change was not affected by lesions in the auditory cortex when the discrimination cue was preserved. However, performance was impaired by lesions in the auditory cortex when a new discrimination cue was used following procedure change. Lesions in the auditory cortex also impaired multimodal discrimination between sound and sound plus light. The present findings suggest that the auditory cortex plays a role as a sensory interface of the higher cortices required for flexible learning and multimodal discrimination.

Acoustic Stimulation↗

Endogenous fluorescence imaging of somatosensory cortical activities after discrimination learning in rats.

Aerobic energy metabolism in the brain is reflected as changes in the green fluorescence of mitochondrial flavoproteins, and the activity-dependent changes in endogenous fluorescence are applicable for functional brain imaging. To understand the roles of cortical plasticity in discrimination learning, we used flavoprotein fluorescence imaging to visualize changes of neural activities in the rat primary somatosensory cortex (SI) after learning. Rats were trained to discriminate floor vibration at rewarded and unrewarded frequencies. After this discrimination learning was accomplished in 3-5 days, the rats were anesthetized with urethane (1.5 g/kg, i.p.), and neural responses were recorded in SI during flutter stimuli applied to the contralateral hindpaw. The fluorescence responses to the stimuli at unrewarded frequencies were selectively depressed in the trained rats, which had behaviorally neglected unrewarded stimuli. The depression of cortical responses was not observed in the rats trained with rewarded stimuli only. Therefore, the stimulus-specific depression in SI might explain a part of neural mechanisms underlying discrimination behavior. To reproduce the stimulus-specific depression of cortical responses in anesthetized rats, tetanic cortical stimulation was paired with flutter stimulation applied to the hindpaw. Selective depression of fluorescence responses or field potentials in SI was induced by the paired stimulation. Our findings suggest that some intracortical circuits in SI are specifically tuned to and modulated by unrewarded stimuli of a particular frequency while SI neurons are responsive to both of rewarded and unrewarded stimuli. The present results indicate the usefulness of flavoprotein fluorescence imaging for investigating somatosensory cortical plasticity after learning.

Animals↗

Sound sequence discrimination learning is dependent on cholinergic inputs to the rat auditory cortex.

In rat auditory cortex (AC) slices, synaptic potentiation following heterosynaptic stimulation is affected by the stimulus sequence used for induction. It was hypothesized that this sequence-dependent plasticity might be partly involved in the cellular mechanisms underlying sound sequence discrimination. Sequence dependence is abolished by muscarinic receptor antagonists. Therefore, dependence of sound sequence discrimination learning on cholinergic inputs to the rat AC was investigated. Rats were trained to discriminate the sequences of two sound components and a licking behavior in response to one of two possible sequences was rewarded with water. Atropine, a muscarinic receptor antagonist, attenuated sound sequence discrimination learning. The acquired sound sequence discrimination was not affected by atropine. Injections of the cholinergic immunotoxin 192IgG-saporin into the AC suppressed sound sequence discrimination learning, while discrimination between the two sound components was not affected. An inhibitor of M-current, linopirdine, restores the sequence dependence of synaptic potentiation in the AC slices suppressed by atropine. In this study, sound sequence discrimination learning attenuated by 192IgG-saporin was also restored by linopirdine. These similarities between sequence dependent plasticity in the AC slices and sound sequence discrimination learning support the hypothesis that the former is involved in the cellular mechanisms underlying the latter.

Acetylcholine↗

Activity-dependent persisting modification of polysynaptic neural circuits involving layer V pyramidal neurons in rat auditory cortex in vitro.

Synaptic plasticity in polysynaptic neural circuits permits modulation of the dynamic properties of these circuits. We investigated the properties of polysynaptic potentiation in pyramidal neurons in layer V of rat auditory cortex (AC) slices using the perforated patch clamp technique. The GABAA receptor inhibitor bicuculline was used to facilitate polysynaptic activity. The amplitude and duration of the polysynaptic activity were both gradually potentiated with repetitive stimulation (RS) at 12 s intervals. Potentiation was saturated within 10 min of the onset of RS. After the cessation of RS, the polysynaptic responses returned to control levels within 30 min. RS-induced potentiation was confirmed by fluorescence imaging of slices loaded with the Ca2+ indicator rhod-2. Such potentiation was not induced by stimulation at 60 s intervals. The magnitude of the RS-induced potentiation in layer V pyramidal neurons in the AC was greater than that in either layer II/III pyramidal neurons in the AC or layer V pyramidal neurons in the visual cortex. The NMDA receptor antagonist APV (100 microm), inhibited RS-induced potentiation. When stimulated at 1 Hz, the potentiated response appeared rapidly. In the absence of bicuculline, RS consisting of five pulses at 30 ms intervals, repeated at 12 s intervals for 10 min, elicited potentiation of firing activity, suggesting that the potentiation is independent of bicuculline. The present study demonstrates the dynamic properties of polysynaptic circuits involving layer V pyramidal neurons in the AC are strongly affected by activity-dependent synaptic potentiation.

Animals↗

Short-term plasticity visualized with flavoprotein autofluorescence in the somatosensory cortex of anaesthetized rats.

In the present study, short-term plasticity of somatosensory neural responses was investigated using flavoprotein autofluorescence imaging in rats anaesthetized with urethane (1.5 g/kg, i.p.) Somatosensory neural activity was elicited by vibratory skin stimulation (50 Hz for 1 s) applied on the surface of the left plantar hindpaw. Changes in green autofluorescence (lambda = 500-550 nm) in blue light (lambda = 450-490 nm) were elicited in the right somatosensory cortex. The normalised maximal fluorescence responses (deltaF/F) was 2.0 +/- 0.1% (n = 40). After tetanic cortical stimulation (TS), applied at a depth of 1.5-2.0 mm from the cortical surface, the responses elicited by peripheral stimulation were significantly potentiated in both peak amplitude and size of the responsive area (both P < 0.02; Wilcoxon signed rank test). This potentiation was clearly observed in the recording session started 5 min after the cessation of TS, and returned to the control level within 30 min. However, depression of the responses was observed after TS applied at a depth of 0.5 mm. TS-induced changes in supragranular field potentials in cortical slices showed a similar dependence on the depth of the stimulated sites. When TS was applied on the ipsilateral somatosensory cortex, marked potentiation of the ipsilateral responses and slight potentiation of the contralateral responses to peripheral stimulation were observed after TS, suggesting the involvement of commissural fibers in the changes in the somatosensory brain maps. The present study clearly demonstrates that functional brain imaging using flavoprotein autofluorescence is a useful technique for investigating neural plasticity in vivo.

Anesthesia↗

Polysynaptic slow depolarization and spiking activity elicited after induction of long-term potentiation in rat auditory cortex.

Polysynaptic activity was recorded in supragranular pyramidal neurons before and after the induction of long-term potentiation (LTP) in slices obtained from rat auditory cortex. LTP was induced by tetanic stimulation of layer IV. In the pyramidal neurons exhibiting LTP, repetitive stimulation at 50 Hz with 15 pulses triggered a slow 15-35 mV depolarization lasting 0.5-2 s with two to five spike discharges. There was no such response before the induction of LTP or in the neurons that did not exhibit LTP. Slow depolarization with spike discharges was blocked by an NMDA receptor antagonist but not by a metabotropic glutamate receptor antagonist. The reversal potential of the slow depolarization was approximately -7 mV and the membrane resistance decreased during slow depolarization, suggesting that the slow depolarization was produced by polysynaptic excitatory post-synaptic potentials. LTP was also induced by low frequency stimulation paired with a depolarizing current injection. In the pyramidal neurons exhibiting LTP after the paired stimulation, the slow depolarization amplitude was small and repetitive stimulation did not trigger spike discharges. Tetanic stimulation is expected to induce LTP in the polysynaptic neural circuits connecting many pyramidal neurons. The present findings suggest that polysynaptic activity can be generated in the potentiated neural circuits. Such activity might serve to read out the memory stored in polysynaptic neural circuits in the cerebral cortex.

Action Potentials↗

Relational discrimination learning between amplitude-modulated sounds in the rat.

We studied the cue used by rats to discriminate between amplitude-modulated (AM) sounds. Water-deprived rats were exposed to two sounds, and licking a spout during the presentation of one of the two (S+) was rewarded with water, while licking during the other (S-) was not. The AM depth was 100, 60 or 20%. Rats were trained to discriminate between 100 and 60% AM sounds over 2 consecutive days. On the next day, the performance in discriminating between 60 and 20% AM sounds was evaluated. Of the four possible combinations of stimuli, a good performance was observed only when the relationship between S+ and S- regarding the AM depth was kept constant throughout the experiments, indicating that the relationship regarding the AM depth was the cue used by the rats for the discrimination.

Acoustic Stimulation↗

Dynamic imaging of somatosensory cortical activity in the rat visualized by flavoprotein autofluorescence.

We used autofluorescence of mitochondrial flavoproteins to image cortical neural activity in the rat. Green autofluorescence in blue light was examined in slices obtained from rat cerebral cortex. About half of the basal autofluorescence was modulated by the presence or absence of O2 or glucose in the medium. Repetitive electrical stimulation at 20 Hz for 1 s produced a localized fluorescence increase in the slices. The amplitude of the increase was 27 +/- 2 % (mean +/- S.D., n = 35). Tetrodotoxin or diphenyleneiodonium, an inhibitor of flavoproteins, blocked the autofluorescence responses. The autofluorescence responses were not observed in slices perfused with calcium-, glucose- or O2-free medium. In the primary somatosensory cortex of rats anaesthetized with urethane (1.5 g kg-1, I.P.), an activity-dependent increase in autofluorescence of 20 +/- 4 % (n = 6) was observed after electrical cortical stimulation at 100 Hz for 1 s, and an increase of 2.6 +/- 0.5 % (n = 33) after vibratory skin stimulation at 50 Hz for 1 s applied to the plantar hindpaw. These responses were large enough to allow visualization of the neural activity without having to average a number of trials. The distribution of the fluorescence responses after electrical or vibratory skin stimulation was comparable to that of the cortical field potentials in the same rats. The fluorescence responses were followed by an increase in arterial blood flow. The former were resistant to an inhibitor of nitric oxide synthase, while the latter was inhibited. Thus, activity-dependent changes in the autofluorescence of flavoproteins are useful for functional brain imaging in vivo.

Anesthesia↗

Anisotropic functional connections between the auditory cortex and area 18a in rat cerebral slices.

We developed a new method to visualize the myeloarchitecture in fresh slices, and investigated the properties of the functional neural connections around the boundary between the primary auditory cortex (area 41) and area 18a in rat cerebral slices. A fresh slice illuminated by near-vertical light was observed with a CCD camera. The translucent images of the slice showed contrast patterns very similar to myeloarchitecture. The boundary between these areas was identified by the well-developed layer IV/V in area 41 but not in area 18a. Antidromic/presynaptic components of the field potentials stimulated and recorded across the areal boundary showed symmetric distribution, while the postsynaptic field potentials in the direction from area 41 to 18a were more prominent than those in the opposite direction in layer II/III. In contrast, the dominant direction of propagation of postsynaptic potentials was from area 18a to 41 in layer V. In the presence of 1 microM bicuculline, an inhibitor of GABA(A) receptors, the polysynaptic activities propagating from area 18a into 41 via layer V were elicited by stimulation of area 18a. The propagation measured by Ca(2+) imaging or field potential recordings was potentiated after both areas 18a and 41 were alternately stimulated several times.

Animals↗

NO enhances presynaptic currents during cerebellar mossy fiber-granule cell LTP.

Nitric oxide (NO) is a candidate retrograde messenger in long-term potentiation (LTP). The NO metabolic pathway is expressed in the cerebellar granule cell layer but its physiological role remained unknown. In this paper we have investigated the role of NO in cerebellar mossy fiber-granule cell LTP, which has postsynaptic N-methyl-d-aspartate (NMDA) receptor-dependent induction. Pre- and postsynaptic current changes were simultaneously measured by using extracellular focal recordings, and NO release was monitored with an electrochemical probe in P21 rat cerebellar slices. High-frequency mossy fiber stimulation induced LTP and caused a significant NO release (6.2 +/- 2.8 nM; n = 5) in the granular layer that was dependent on NMDA receptor as well as on nitric oxide synthase (NOS) activation. Preventing NO production by perfusing the NOS inhibitor 100 microM NG-nitro-l-arginine (L-NNA), blocking extracellular NO diffusion by 10 microM MbO2, or inhibiting the NO target guanylyl cyclase (sGC) with 10 microM 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-dione (ODQ) prevented LTP. Moreover, the NO donor 10 microM 2-(N,N-diethylamino)-diazenolate-2-oxide.Na (DEA-NO) induced LTP, which was mutually occlusive with LTP generated by high-frequency stimulation, prevented by ODQ, and insensitive to NMDA channel blockade (50 microM APV + 25 microM 7-Cl-kyn) or interruption of mossy fiber stimulation. Thus NO is critical for LTP induction at the cerebellar mossy fiber-granule cell relay. Interestingly, LTP manipulations were accompanied by consensual changes in the presynaptic current, suggesting that NO acts as a retrograde signal-enhancing presynaptic terminal excitability.

Animals↗