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Animals optimize their actions by predicting and verifying their outcomes (e.g., rewards). Reward prediction often requires working memory (WM)-based information. To elucidate the neural basis of WM-based reward prediction, we compared the activity of dopamine (DA) neurons in the ventral tegmental area (VTA) in an alternate reward condition (WM-based) with that in a random (WM-free) reward condition in rats and mice. Positron emission tomography revealed greater VTA activation in the WM-based than the WM-free condition. Lateral and medial VTA neurons displayed differential electrophysiological spike activities reflecting WM-based and WM-free reward prediction error, respectively. Furthermore, phasic DA release in the dorsal and ventral striatum changed as WM-based and WM-free classical conditioning progressed. Consistent with our WM-based model, reward acquisition caused a DA dip only in the dorsal striatum. Thus a dual DA system processes WM-based and WM-free reward prediction in parallel.

Cerebellar climbing fibers (CFs) convey sensorimotor information and their errors, which are used for motor control and learning. Furthermore, they represent reward-related information. Despite such functional diversity of CF signals, it is still unclear whether each CF conveys the information of single or multiple modalities and how the CFs conveying different information are distributed over the cerebellar cortex. We performed two-photon calcium imaging from cerebellar Purkinje cells (PCs) in mice engaged in a voluntary forelimb lever-pull task and demonstrated that CF responses in 93% of PCs could be explained by the combination of multiple behavioral variables, such as lever movement, licking, and reward delivery. Neighboring PCs exhibited similar CF response properties, formed functional clusters, and shared noise fluctuations of responses. Taken together, individual CFs convey behavioral information on multiplex variables and are spatially organized into the functional modules of the cerebellar cortex.

Optogenetic tools such as channelrhodopsin-2 (ChR2) enable the manipulation and mapping of neural circuits. However, ChR2 variants selectively transported down a neuron’s long-range axonal projections for precise presynaptic activation remain lacking. As a result, ChR2 activation is often contaminated by the spurious activation of en passant fibers that compromise the accurate interpretation of functional effects. Here, we explored the engineering of a ChR2 variant specifically localized to presynaptic axon terminals. The metabotropic glutamate receptor 2 (mGluR2) C-terminal domain fused with a proteolytic motif and axon-targeting signal (mGluR2-PA tag) localized ChR2-YFP at axon terminals without disturbing normal transmission. mGluR2-PA-tagged ChR2 evoked transmitter release in distal projection areas enabling lower levels of photostimulation. Circuit connectivity mapping in vivo with the Spike Collision Test revealed that mGluR2-PA-tagged ChR2 is useful for identifying axonal projection with significant reduction in the polysynaptic excess noise. These results suggest that the mGluR2-PA tag helps actuate trafficking to the axon terminal, thereby providing abundant possibilities for optogenetic experiments., Hamada et al. engineer and utilise a channelrhodopsin-2 variant that is localized to presynaptic axon terminals. They demonstrate its use for circuitry mapping in vivo and thus provide a useful tool for future optogenetic experiments

Standard analysis of neuronal functions assesses the temporal correlation between animal behaviors and neuronal activity by aligning spike trains with the timing of a specific behavioral event, e.g., visual cue. However, spike activity is often involved in information processing dependent on a relative phase between two consecutive events rather than a single event. Nevertheless, less attention has so far been paid to such temporal features of spike activity in relation to two behavioral events. Here, we propose "Phase-Scaling analysis" to simultaneously evaluate the phase locking and scaling to the interval between two events in task-related spike activity of individual neurons. This analysis method can discriminate conceptual "scaled"-type neurons from "nonscaled"-type neurons using an activity variation map that combines phase locking with scaling to the interval. Its robustness was validated by spike simulation using different spike properties. Furthermore, we applied it to analyzing actual spike data from task-related neurons in the primary visual cortex (V1), posterior parietal cortex (PPC), primary motor cortex (M1), and secondary motor cortex (M2) of behaving rats. After hierarchical clustering of all neurons using their activity variation maps, we divided them objectively into four clusters corresponding to nonscaled-type sensory and motor neurons and scaled-type neurons including sustained and ramping activities, etc. Cluster/subcluster compositions for V1 differed from those of PPC, M1, and M2. The V1 neurons showed the fastest functional activities among those areas. Our method was also applicable to determine temporal "forms" and the latency of spike activity changes. These findings demonstrate its utility for characterizing neurons.

The entorhinal cortex (EC) is the main interface between the hippocampus and the neocortex. The EC plays a critical role in learning and memory. We investigated the neuronal representation of behavioral events during operant learning in the hippocampal-entorhinal circuit of head-fixed rats. Both CA1 and lateral entorhinal cortex (LEC) neurons develop task-related activities after learning. Among diverse task-related activities, we compared the transient peak activities that represent action and reward and found a distinct difference in the timing of behavioral event representation between CA1 and LEC. CA1 represents action and reward events in close to real-time, whereas both the superficial and deep layers of the LEC showed delayed representation of those events. Our results suggest that subpopulations exist within which CA1 and LEC neurons process the information in a different order from the anatomically defined hippocampal-entorhinal circuit.

The spike collision test is a highly reliable technique to identify axonal projection of a neuron recorded electrophysiologically for investigating functional spike information among brain areas. It is potentially applicable to more neuronal projections by combining multi-channel recording with optogenetic stimulation. Yet, it remains inefficient and laborious because an experimenter must visually select spikes in every channel and manually repeat spike collision tests for each neuron serially. Here, we established a novel technique to automatically perform spike collision tests for all channels in parallel (Multi-Linc analysis), employing two distinct protocols implemented in a multi-channel real-time processing system. The rat cortical neurons identified with this technique displayed physiological spike features consistent with excitatory projection neurons. Their antidromic spikes were similar in shape but slightly larger in amplitude compared with spontaneous spikes. In addition, we demonstrated simultaneous identification of reciprocal or bifurcating projections among cortical areas. Thus, our Multi-Linc analysis will be a powerful research approach to elucidate interareal spike communication.

The cerebral cortex usually governs contralateral body parts in sensation and movements. The rule of contralateral dominance of cerebral cortex is well established on the basis of a long history of human and animal experiments. We also confirmed the rule for the primary (M1) and secondary (M2) motor cortices controlling unilateral forelimb movements in behaving rats. However, we found that their posterior parietal cortex (PPC) neurons preferentially represent ipsilateral forelimb movements, in contrast to the contralateral preference of M1 and M2 neurons. Moreover, our optogenetic activation of PPC neurons evoked ipsilaterally biased forelimb movements. Even weak PPC activation affected their task performance of ipsilateral forelimb movements. I will talk about our interpretation on these paradoxical observations from the point of view of an evolutional difference between rodents and primates.

It is well known that the posterior parietal cortex (PPC) and frontal motor cortices in primates preferentially control voluntary movements of contralateral limbs. The PPC of rats has been defined based on patterns of thalamic and cortical connectivity. The anatomical characteristics of this area suggest that it may be homologous to the PPC of primates. However, its functional roles in voluntary forelimb movements have not been well understood, particularly in the lateralization of motor limb representation; that is, the limb-specific activity representations for right and left forelimb movements. We examined functional spike activity of the PPC and two motor cortices, the primary motor cortex (M1) and the secondary motor cortex (M2), when head-fixed male rats performed right or left unilateral movements. Unlike primates, PPC neurons in rodents were found to preferentially represent ipsilateral forelimb movements, in contrast to the contralateral preference of M1 and M2 neurons. Consistent with these observations, optogenetic activation of PPC and motor cortices, respectively, evoked ipsilaterally and contralaterally biased forelimb movements. Finally, we examined the effects of optogenetic manipulation on task performance. PPC or M1 inhibition by optogenetic GABA release shifted the behavioral limb preference contralaterally or ipsilaterally, respectively. In addition, weak optogenetic PPC activation, which was insufficient to evoke motor responses by itself, shifted the preference ipsilaterally; although similar M1 activation showed no effects on task performance. These paradoxical observations suggest that the PPC plays evolutionarily different roles in forelimb control between primates and rodents.SIGNIFICANCE STATEMENTIn rodents, the primary and secondary motor cortices (M1 and M2, respectively) are involved in voluntary movements with contralateral preference. However, it remains unclear whether and how the posterior parietal cortex (PPC) participates in controlling multiple limb movements. We recorded functional activity from these areas using a behavioral task to monitor movements of the right and left forelimbs separately. PPC neurons preferentially represented ipsilateral forelimb movements and optogenetic PPC activation evoked ipsilaterally biased forelimb movements. Optogenetic PPC inhibition via GABA release shifted the behavioral limb preference contralaterally during task performance, whereas weak optogenetic PPC activation, which was insufficient to evoke motor responses by itself, shifted the preference ipsilaterally. Our findings suggest rodent PPC contributes to ipsilaterally biased motor response and/or planning.

Animals can suppress their behavioral response in advance according to changes in environmental context (proactive inhibition: delaying the start of response), a process in which several cortical areas may participate. However, it remains unclear how this process is adaptively regulated according to contextual changes on different timescales. To address the issue, we used an improved stop-signal task paradigm to behaviorally and electrophysiologically characterize the temporal aspect of proactive inhibition in head-fixed rats. In the task, they must respond to a go cue as quickly as possible (go trial), but did not have to respond if a stop cue followed the go cue (stop trial). The task alternated between a block of only go trials (G-block) and a block of go-and-stop trials (GS-block). We observed block-based and trial-based proactive inhibition (emerging in GS-block and after stop trial, respectively) by behaviorally evaluating the delay in reaction time in correct go trials depending on contextual changes on different timescales. We electrophysiologically analyzed task-related neuronal activity in the primary and secondary motor, posterior parietal, and orbitofrontal cortices (M1, M2, PPC, and OFC, respectively). Under block-based proactive inhibition, spike activity of cue-preferring OFC neurons was attenuated continuously, while M1 and M2 activity was enhanced during motor preparation. Subsequently, M1 activity was attenuated during motor decision/execution. Under trial-based proactive inhibition, the OFC activity was continuously enhanced, and PPC and M1 activity was also enhanced shortly during motor decision/execution. These results suggest that different cortical mechanisms underlie the two types of proactive inhibition in rodents.

Neurons in medial frontal cortex (MFC) receive sensory signals that are crucial for decision-making behavior. While decision-making is easy for familiar sensory signals, it becomes more elaborative when sensory signals are less familiar to animals. It remains unclear how the population of neurons enables the coordinate transformation of such a sensory input into ambiguous choice responses. Furthermore, whether and how cortical oscillations temporally coordinate neuronal firing during this transformation has not been extensively studied. Here, we recorded neuronal population responses to familiar or unfamiliar auditory cues in rat MFC and computed their probabilistic evolution. Population responses to familiar sounds organize into neuronal trajectories containing multiplexed sensory, motor, and choice information. Unfamiliar sounds, in contrast, evoke trajectories that travel under the guidance of familiar paths and eventually diverge to unique decision states. Local field potentials exhibited beta- (15-20 Hz) and gamma-band (50-60 Hz) oscillations to which neuronal firing showed modest phase locking. Interestingly, gamma oscillation, but not beta oscillation, increased its power abruptly at some timepoint by which neural trajectories for different choices were near maximally separated. Our results emphasize the importance of the evolution of neural trajectories in rapid probabilistic decisions that utilize unfamiliar sensory information.

In motor cortex, 2 types of deep layer pyramidal cells send their axons to other areas: intratelencephalic (IT)-type neurons specifically project bilaterally to the cerebral cortex and striatum, whereas neurons of the extratelencephalic (ET)-type, termed conventionally pyramidal tract-type, project ipsilaterally to the thalamus and other areas. Although they have totally different synaptic and membrane potential properties in vitro, little is known about the differences between them in ongoing spiking dynamics in vivo. We identified IT-type and ET-type neurons, as well as fast-spiking-type interneurons, using novel multineuronal analysis based on optogenetically evoked spike collision along their axons in behaving/resting rats expressing channelrhodopsin-2 (Multi-Linc method). We found "postspike suppression" (~100 ms) as a characteristic of ET-type neurons in spike auto-correlograms, and it remained constant independent of behavioral conditions in functionally different ET-type neurons. Postspike suppression followed even solitary spikes, and spike bursts significantly extended its duration. We also observed relatively strong spike synchrony in pairs containing IT-type neurons. Thus, spiking dynamics in IT-type and ET-type neurons may be optimized differently for precise and coordinated motor control.

Key points There have been few systematic population-wide analyses of relationships between spike synchrony within a period of several milliseconds and behavioural functions. In this study, we obtained a large amount of spike data from > 23,000 neuron pairs by multiple single-unit recording from deep layer neurons in motor cortical areas in rats performing a forelimb movement task. The temporal changes of spike synchrony in the whole neuron pairs were statistically independent of behavioural changes during the task performance, although some neuron pairs exhibited correlated changes in spike synchrony. Mutual information analyses revealed that spike synchrony made a smaller contribution than spike rate to behavioural functions. The strength of spike synchrony between two neurons was statistically independent of the spike rate-based preferences of the pair for behavioural functions. Abstract Spike synchrony within a period of several milliseconds in presynaptic neurons enables effective integration of functional information in the postsynaptic neuron. However, few studies have systematically analysed the population-wide relationships between spike synchrony and behavioural functions. Here we obtained a sufficiently large amount of spike data among regular-spiking (putatively excitatory) and fast-spiking (putatively inhibitory) neuron subtypes (> 23,000 pairs) by multiple single-unit recording from deep layers in motor cortical areas (caudal forelimb area, rostral forelimb area) in rats performing a forelimb movement task. After holding a lever, rats pulled the lever either in response to a cue tone (external-trigger trials) or spontaneously without any cue (internal-trigger trials). Many neurons exhibited functional spike activity in association with forelimb movements, and the preference of regular-spiking neurons in the rostral forelimb area was more biased toward externally triggered movement than that in the caudal forelimb area. We found that a population of neuron pairs with spike synchrony does exist, and that some neuron pairs exhibit a dependence on movement phase during task performance. However, the population-wide analysis revealed that spike synchrony was statistically independent of the movement phase and the spike rate-based preferences of the pair for behavioural functions, whereas spike rates were clearly dependent on the movement phase. In fact, mutual information analyses revealed that the contribution of spike synchrony to the behavioural functions was small relative to the contribution of spike rate. Our large-scale analysis revealed that cortical spike rate, rather than spike synchrony, contributes to population coding for movement.

STUDY OBJECTIVES The sleep spindle has been implicated in thalamic sensory gating, cortical development, and memory consolidation. These multiple functions may depend on specific spatiotemporal emergence and interactions with other spindles and other forms of brain activity. Therefore, we measured sleep spindle cortical distribution, regional heterogeneity, synchronization, and phase relationships with other electroencephalographic components in freely moving primates. METHODS Transcortical field potentials were recorded from Japanese monkeys via telemetry and were analyzed using the Hilbert-Huang transform. RESULTS Spindle (12-20 Hz) current sources were identified over a wide region of the frontoparietal cortex. Most spindles occurred independently in their own frequency, but some appeared concordant between cortical areas with frequency interdependence, particularly in nearby regions and bilaterally symmetrical regions. Spindles in the dorsolateral prefrontal cortex appeared around the surface-positive and depth-negative phase of transcortically recorded slow oscillations (< 1 Hz), whereas centroparietal spindles emerged around the opposite phase. The slow-oscillation phase reversed between the prefrontal and central regions. Gamma activities increased before spindle onset. Several regional heterogeneities in properties of human spindles were replicated in the monkeys, including frequency, density, and inter-cortical time lags, although their topographic patterns were different from those of humans. The phase-amplitude coupling between spindle and gamma activity was also replicated. CONCLUSIONS Spindles in widespread cortical regions are possibly driven by independent rhythm generators, but are temporally associated to spindles in other regions and to slow and gamma oscillations by corticocortical and thalamocortical pathways.

A feature of information processing in the brain is to have the ability of integrating novelty information into past experience. In other words, this integration ability is to manipulate spatiotemporal context information in learning and memory. In this paper, we propose a neural network model, which accounts for this feature, and show the possibility to acquire a spatiotemporal context information by the coding method of spatial clustering and its self-similarity in learning and memory based on the hippocampal experimental data. We also consider the physiological validity of this model.

Each area in the brain sends functional spike outputs to other areas through axonal projections. Until now, however, it has been technically difficult to precisely measure the spike outputs among brain areas in behaving animals. We recently established the Multi-Linc analysis method, which uses optogenetically evoked spike collision in multineuronal recording, to elucidate inter-areal spike communication at a single-cell level at millisecond time resolution. Here, I review the concept and demonstration of this method and discuss its advantages and future directions.

In the parkinsonian state, the motor cortex and basal ganglia (BG) undergo dynamic remodeling of movement representation. One such change is the loss of the normal contralateral lateralized activity pattern. The increase in the number of movement-related neurons responding to ipsilateral or bilateral limb movements may cause motor problems, including impaired balance, reduced bimanual coordination, and abnormal mirror movements. However, it remains unknown how individual types of motor cortical neurons organize this reconstruction. To explore the effect of dopamine depletion on lateralized activity in the parkinsonian state, we used a partial hemiparkinsonian model [6-hydroxydopamine (6-OHDA) lesion] in Long–Evans rats performing unilateral movements in a right–left pedal task, while recording from primary (M1) and secondary motor cortex (M2). The lesion decreased contralateral preferred activity in both M1 and M2. In addition, this change differed among identified intratelencephalic (IT) and pyramidal tract (PT) cortical projection neurons, depending on the cortical area. We detected a decrease in lateralized activity only in PT neurons in M1, whereas in M2, this change was observed in IT neurons, with no change in the PT population. Our results suggest a differential effect of dopamine depletion in the lateralized activity of the motor cortex, and suggest possible compensatory changes in the contralateral hemisphere.

Recent studies have shown that the dendrites of several neurons are not simple translators but are crucial facilitators of excitatory postsynaptic potential (EPSP) propagation and summation of synaptic inputs to compensate for inherent voltage attenuation. Granule cells (GCs)are located at the gateway for valuable information arriving at the hippocampus from the entorhinal cortex. However, the underlying mechanisms of information integration along the dendrites of GCs in the hippocampus are still unclear. In this study, we investigated the input integration around dendritic branches of GCs in the rat hippocampus. We applied differential spatiotemporal stimulations to the dendrites using a high-speed glutamate-uncaging laser. Our results showed that when two sites close to and equidistant from a branching point were simultaneously stimulated, a nonlinear summation of EPSPs was observed at the soma. In addition, nonlinear summation (facilitation) depended on the stimulus location and was significantly blocked by the application of a voltage-dependent Ca(2+) channel antagonist. These findings suggest that the nonlinear summation of EPSPs around the dendritic branches of hippocampal GCs is a result of voltage-dependent Ca(2+) channel activation and may play a crucial role in the integration of input information.

Sequential motor behavior requires a progression of discrete preparation and execution states. However, the organization of state-dependent activity in neuronal ensembles of motor cortex is poorly understood. Here, we recorded neuronal spiking and local field potential activity from rat motor cortex during reward-motivated movement and observed robust behavioral state-dependent coordination between neuronal spiking, γ oscillations, and θ oscillations. Slow and fast γ oscillations appeared during distinct movement states and entrained neuronal firing. γ oscillations, in turn, were coupled to θ oscillations, and neurons encoding different behavioral states fired at distinct phases of θ in a highly layer-dependent manner. These findings indicate that θ and nested dual band γ oscillations serve as the temporal structure for the selection of a conserved set of functional channels in motor cortical layer activity during animal movement. Furthermore, these results also suggest that cross-frequency couplings between oscillatory neuronal ensemble activities are part of the general coding mechanism in cortex.

In order to make a direct link between the morphological and functional study of the nervous system, we established an experimental protocol for labeling individual neurons persistently without microscopic guidance by injecting a plasmid encoding fluorescent protein electroporatively after recording their activity extracellularly. Using a glass pipette filled with electrolyte solution containing a plasmid encoding green fluorescent protein (GFP), single-neuron recording and electroporation were performed on anesthetized rats. When performing the electroporation at the completion of recording, the degree of contact between the target neuron and the electrode tip was adjusted by monitoring the change of the trace of recorded action potentials and the increase of electrode resistance. The expression of GFP and its immunostaining with a polyclonal antibody enabled us to clearly see the basic structural components such as cell bodies, axons, dendrites, and even smaller components such as spines. Identification of the morphological subtypes of neurons was possible with every labeled neuron. The optimum condition for labeling was a 30% increase of the electrode resistance, and the labeling success rate evaluated 3 days after labeling was 40%. The rate evaluated one month after labeling was only slightly lower (33%). We also confirmed experimentally that this recording and labeling procedure can be similarly successful in head-fixed behaving rats. This new experimental protocol will be a breakthrough in systems neuroscience because it makes a direct link between the morphology and behavior-related activity of single neurons.

Theoretical simulations suggest that spike rate is regulated by varying both membrane potential and its fluctuation. We investigated whether membrane potential fluctuation functionally changes in motor cortex and striatum neurons during discrete forelimb movements and pauses, or at rest, using whole-cell recording in task-performing rats. Membrane potential fluctuation was diminished by task performance, but maintained overall in the alpha/beta and gamma bands during forelimb movements and pauses. By contrast, membrane potential itself was correlated with spike rate in task-related neurons. Thus, membrane potential, but not its fluctuation, is a critical determinant of execution and pausing of discrete movements.

Functional clustering of neurons is frequently observed in the motor cortex. However, it is unknown if, when, and how fine-scale (3 s to pull it again. We found two types of task-related cells in the mice: cells whose peak activities occurred during lever pulls (pull cells) and cells whose peak activities occurred after the end of lever pulls. The activity of pull cells was strongly associated with lever-pull duration. In ∼40% of imaged fields, functional clusterings were temporally detected during the lever pulls. Spatially, there were ∼70-μm-scale clusters that consisted of more than four pull cells in ∼50% of the fields. Ensemble and individual activities of pull cells within the cluster more accurately predicted lever movement trajectories than activities of pull cells outside the cluster. This was likely because clustered pull cells were more often active in the individual trials than pull cells outside the cluster. This higher fidelity of activity was related to higher trial-to-trial correlations of activities of pairs within the cluster. We propose that strong recurrent network clusters may represent the execution of voluntary movements.

Summary The basal ganglia play key roles in adaptive behaviors guided by reward and punishment. However, despite accumulating knowledge, few studies have tested how heterogeneous signals in the basal ganglia are organized and coordinated for goal-directed behavior. In this study, we investigated neuronal signals of the direct and indirect pathways of the basal ganglia as rats performed a lever push/pull task for a probabilistic reward. In the dorsomedial striatum, we found that optogenetically and electrophysiologically identified direct pathway neurons encoded reward outcomes, whereas indirect pathway neurons encoded no-reward outcome and next-action selection. Outcome coding occurred in association with the chosen action. In support of pathway-specific neuronal coding, light activation induced a bias on repeat selection of the same action in the direct pathway, but on switch selection in the indirect pathway. Our data reveal the mechanisms underlying monitoring and updating of action selection for goal-directed behavior through basal ganglia circuits.

Simultaneous recordings with multi-channel electrodes are widely used for studying how multiple neurons are recruited for information processing. The recorded signals contain the spike events of a number of adjacent or distant neurons and must be sorted correctly into spike trains of individual neurons. Several mathematical methods have been proposed for spike sorting but the process is difficult in practice, as extracellularly recorded signals are corrupted by biological noise. Moreover, spike sorting is often time-consuming, as it usually requires corrections by human operators. Methods are needed to obtain reliable spike clusters without heavy manual operation. Here, we introduce several methods of spike sorting and compare the accuracy and robustness of their performance by using publicized data of simultaneous extracellular and intracellular recordings of neuronal activity. The best and excellent performance was obtained when a newly proposed filter for spike detection was combined with the wavelet transform and variational Bayes for a finite mixture of Student's t-distributions, namely, robust variational Bayes. Wavelet transform extracts features that are characteristic of the detected spike waveforms and the robust variational Bayes categorizes the extracted features into clusters corresponding to spikes of the individual neurons. The use of Student's t-distributions makes this categorization robust against noisy data points. Some other new methods also exhibited reasonably good performance. We implemented all of the proposed methods in a C++ code named 'EToS' (Efficient Technology of Spike sorting), which is freely available on the Internet.

The hippocampus is a remarkable neural structure that displays a variety of synchronous oscillations that may be physiological or pathophysiological, such as theta rhythms and epileptic seizures. Electrically induced seizure-like afterdischarges are an excellent system for elucidating the network mechanisms underlying neuronal synchronization and rhythm generation of epileptic synchronous oscillations in extremely hyperactive hippocampal networks. In this Update Article, we review key findings of studies on these electrically induced seizure-like afterdischarges in vitro. During these afterdischarges, GABAergic responses become transiently depolarizing and even excitatory as chloride rapidly accumulates postsynaptically in pyramidal cells. Glutamate and potassium enhance this transient GABAergic excitation. Neuronal synchronization of afterdischarge is achieved by GABAergic and glutamatergic excitation of pyramidal cells and interneurons localized in the stratum pyramidale and stratum oriens. Rhythm generation in seizure-like synchronous oscillations is not yet understood but is the subject of intensive study.

The architectonic subdivisions of the brain are believed to be functional modules, each processing parts of global functions. Previously, we showed that neurons in different regions operate in different firing regimes in monkeys. It is possible that firing regimes reflect differences in underlying information processing, and consequently the firing regimes in homologous regions across animal species might be similar. We analyzed neuronal spike trains recorded from behaving mice, rats, cats, and monkeys. The firing regularity differed systematically, with differences across regions in one species being greater than the differences in similar areas across species. Neuronal firing was consistently most regular in motor areas, nearly random in visual and prefrontal/medial prefrontal cortical areas, and bursting in the hippocampus in all animals examined. This suggests that firing regularity (or irregularity) plays a key role in neural computation in each functional subdivision, depending on the types of information being carried.SIGNIFICANCE STATEMENTBy analyzing neuronal spike trains recorded from mice, rats, cats, and monkeys, we found that different brain regions have intrinsically different firing regimes that are more similar in homologous areas across species than across areas in one species. Because different regions in the brain are specialized for different functions, the present finding suggests that the different activity regimes of neurons are important for supporting different functions, so that appropriate neuronal codes can be used for different modalities.

Temporal integration of externally or internally driven information is required for a variety of cognitive processes. This computation is generally linked with graded rate changes in cortical neurons, which typically appear during a delay period of cognitive task in the prefrontal and other cortical areas. Here, we present a neural network model to produce graded (climbing or descending) neuronal activity. Model neurons are interconnected randomly by AMPA-receptor–mediated fast excitatory synapses and are subject to noisy background excitatory and inhibitory synaptic inputs. In each neuron, a prolonged afterdepolarizing potential follows every spike generation. Then, driven by an external input, the individual neurons display bimodal rate changes between a baseline state and an elevated firing state, with the latter being sustained by regenerated afterdepolarizing potentials. When the variance of background input and the uniform weight of recurrent synapses are adequately tuned, we show that stochastic noise and reverberating synaptic input organize these bimodal changes into a sequence that exhibits graded population activity with a nearly constant slope. To test the validity of the proposed mechanism, we analyzed the graded activity of anterior cingulate cortex neurons in monkeys performing delayed conditional Go/No-go discrimination tasks. The delay-period activities of cingulate neurons exhibited bimodal activity patterns and trial-to-trial variability that are similar to those predicted by the proposed model.

It has recently been shown that electrical stimulation in normal extracellular fluid induces seizure-like afterdischarge activity that is always preceded by GABA-dependent slow depolarization. These afterdischarge responses are synchronous among mature hippocampal neurons and driven by excitatory GABAergic input. However, the differences in the mechanisms whereby the GABAergic signals in pyramidal cells and interneurons are transiently converted from hyperpolarizing to depolarizing (and even excitatory) have remained unclear. To clarify the network mechanisms underlying this rapid GABA conversion that induces afterdischarges, we examined the temporal changes in GABAergic responses in pyramidal cells and/or interneurons of the rat hippocampal CA1 area in vitro. The extents of slow depolarization and GABA conversion were much larger in the pyramidal cell group than in any group of interneurons. Besides GABA(A) receptor activation, neuronal excitation by ionotropic glutamate receptors enhanced GABA conversion in the pyramidal cells and consequent induction of afterdischarge. The slow depolarization was confirmed to consist of two distinct phases; an early phase that depended primarily on GABA(A)-mediated postsynaptic Cl- accumulation, and a late phase that depended on extracellular K+ accumulation, both of which were enhanced by glutamatergic neuron excitation. Moreover, extracellular K+ accumulation augmented each oscillatory response of the afterdischarge, probably by further Cl- accumulation through K+-coupled Cl- transporters. Our findings suggest that the GABA reversal potential may be elevated above their spike threshold predominantly in the pyramidal cells by biphasic Cl- intrusion during the slow depolarization in GABA- and glutamate-dependent fashion, leading to the initiation of seizure-like epileptiform activity.

Recent evidence has suggested that theta-frequency (4–7 Hz) oscillations around the human anterior cingulate cortex (ACC) and frontal cortex—that is, frontal midline theta (Fm theta) oscillations—may be involved in attentional processes in the brain. However, little is known about the physiological basis of Fm theta oscillations because invasive study in the human is allowed in only limited cases. In the present study, we developed a monkey model for Fm theta oscillations and located the generators of theta waves using electrodes implanted in various cortical areas. Monkeys were engaged in a self-initiated hand-movement task with a waiting period. The theta power in area 9 (the medial prefrontal cortex) and area 32 (the rostral ACC) was gradually increased from a few seconds before the movement and reached a peak immediately after the movement. When the movement was rewarded, the theta power attained a second peak, whereas it swiftly decreased in the unrewarded trials. Theta oscillations in areas 9 and 32 were coherent and phase locked together. This theta activity may be associated with “executive attention” including self-control, internal timing, and assessment of reward. It is probably a homologue of human Fm theta oscillations, as judged from the similar localization, corresponding frequency, and dependency on attentional processes. The monkey model would be useful for studying executive functions in the frontal cortex.

It is known that GABA is a major inhibitory neurotransmitter in mature mammalian brains, but the effect of this substance is sometimes converted into depolarizing or even excitatory when the postsynaptic Cl– concentration becomes high. Recently we have shown that seizurelike afterdischarge induced by tetanic stimulation in normal extracellular fluid (posttetanic afterdischarge) is mediated through GABAergic excitation in mature hippocampal CA1 pyramidal cells. In this study, we examined the possible contribution of similar depolarizing/excitatory GABAergic input to the CA1 pyramidal cells to the seizurelike afterdischarge induced in a low extracellular Mg2+ condition, another experimental model of epileptic seizure activity (low-Mg2+ afterdischarge). Perfusion of the GABAA antagonist bicuculline abolished the low-Mg2+ afterdischarge, but not the interictal-like activity, in most cases. Each oscillatory response during the low-Mg2+ afterdischarge was dependent on Cl– conductance and contained an F–-insensitive depolarizing component in the pyramidal cells, thus indicating that the afterdischarge response may be mediated through both GABAergic and nonGABAergic transmissions. In addition, local GABA application to the recorded cells revealed that GABA responses were indeed depolarizing during the low-Mg2+ afterdischarge. Furthermore, the GABAergic interneurons located in the strata pyramidale and oriens fired in oscillatory cycles more actively than those in other layers of the CA1 region. These results suggest that the depolarizing GABAergic input may facilitate oscillatory synchronization among the hippocampal CA1 pyramidal cells during the low-Mg2+ afterdischarge in a manner similar to the expression of the posttetanic afterdischarge.

Strong tetanization induces synchronous membrane potential oscillations (seizure-like afterdischarge) in mature pyramidal cells of the hippocampal CA1 region. To investigate whether local networks in other brain regions can generate such an afterdischarge independently, we studied the inducibility of afterdischarge in individual ‘isolated slices’ of the rat hippocampal CA1 and CA3 regions, dentate gyrus (DG), entorhinal cortex (EC), and temporal cortex (TC) using intracellular and extracellular recordings. The strong tetanization constantly induced afterdischarges in the CA1 and CA3 pyramidal cells as well as in the EC and TC superficial principal cells. However, parameters of the afterdischarges, such as the frequency and duration of afterdischarges, varied among the regions. A mixture of N -methyl- d -aspartate (NMDA) and non-NMDA receptor antagonists or a GABA A receptor antagonist completely blocked the afterdischarges. Local GABA application during the afterdischarge elicited depolarization, rather than hyperpolarization. Moreover, reversal potentials of the afterdischarge were around −40 mV. In contrast, the tetanization resulted in occasional afterdischarge-like activities in DG slices, which were blocked by the non-NMDA or GABA A receptor antagonist. These findings suggest that the afterdischarges mediated through the excitatory GABAergic and glutamatergic transmissions might be common to, but be modulated differently by individual local networks in the hippocampus and cortex.

We have recently reported that excitatory GABAergic and glutamatergic mechanisms may be involved in the generation of seizure-like (ictal) rhythmic synchronization (afterdischarge), induced by a strong synaptic stimulation of the CA1 pyramidal cells in the mature rat hippocampus in vitro. To clarify the network mechanism of this neuronal synchronization, dual whole-cell patch-clamp recordings of the afterdischarge responses were performed simultaneously from a variety of interneurones and their neighbouring pyramidal cells in hippocampal CA1-isolated slice preparations. According to morphological and electrophysiological criteria, the recorded interneurones were then classified into distinct subtypes. The non-fast-spiking interneurones located in the strata lacunosum-moleculare and radiatum hardly discharged during the afterdischarge, whereas most of the fast-spiking and non-fast-spiking interneurones in the strata oriens and pyramidale, including the basket, chandelier and bistratified cells, exhibited prominent firings that were precisely synchronous with oscillatory responses in the pyramidal cells. Field potential recordings showed that excitatory synaptic transmissions might take place primarily in the strata oriens and pyramidale during the afterdischarge. Restricted lesions in the strata oriens and pyramidale, but not in the other layers, resulted in the complete desynchronization of afterdischarge activity, and also, local application of glutamate receptor antagonists to these layers blocked the expression of afterdischarge. The present findings indicate that the neuronal synchronization of epileptic afterdischarge may be accomplished in a ‘positive feedback circuit’ formed by the excitatory GABAergic interneurones and the glutamatergic pyramidal cells within the strata oriens and/or pyramidale of the hippocampal CA1 region.

In order to investigate the functional significance of theta oscillations in the brain, we recorded the cortical field potential in monkeys engaged in a visually-initiated hand movement task. In each trial a warning signal (S1) was followed 3 s later by an imperative signal (S2) to which the monkey had to respond to get a reward. The theta power in the prefrontal area 9 and the prelimbic area 32 was higher in the S1-S2 interval than in the pre-S1 period. This theta activity may be related to attentional processes and is probably a homologue of the human frontal midline theta (Fm theta) rhythms.

It is known that GABA, a major inhibitory transmitter in the CNS, acts as an excitatory (or depolarizing) transmitter transiently after intense GABAA receptor activation in adult brains. The depolarizing effect is considered to be dependent on two GABAA receptor-permeable anions, chloride (Cl–) and bicarbonate (HCO3–). However, little is known about their spatial and temporal profiles during the GABAergic depolarization in postsynaptic neurons. In the present study, we show that the amplitude of synaptically induced depolarizing response was correlated with intracellular Cl– accumulation in the soma of mature hippocampal CA1 pyramidal cells, by using whole cell patch-clamp recording and Cl– imaging technique with a Cl– indicator 6-methoxy- N-ethylquinolinium iodide (MEQ). The synaptically activated Cl– accumulation was mediated dominantly through GABAA receptors. Basket cells, a subclass of fast-spiking interneurons innervating the somatic portion of the pyramidal cells, actually fired at high frequency during the Cl– accumulation accompanying the depolarizing responses. These results suggest synaptically activated GABAA-mediated Cl– accumulation may play a critical role in generation of an excitatory GABAergic response in the mature pyramidal cells receiving intense synaptic inputs. This may be the first demonstration of microscopic visualization of intracellular Cl– accumulation during synaptic activation.

Noninvasive imaging techniques showed that the anterior cingulate cortex is related to higher-order cognitive and motor-related functions in humans. To elucidate the cellular mechanism of such cingulate functions, single-unit activity was recorded from three cingulate motor areas of macaque monkeys performing delayed conditional Go/No-go discrimination tasks using spatial (location) and nonspatial (color) visual cues. Unlike prefrontal neurons, only a few neurons coded the visual information on individual features (e.g., “left” or “red”) in all of the rostral (CMAr), dorsal (CMAd), and ventral (CMAv) cingulate motor areas. Instead, many neurons in the CMAr exhibited the attention-like activity anticipating the second (conditioned) visual cues, with the specificity to visual category (“location” or “color”). In addition, there were a number of CMAr neurons specific to motor response (GoorNo-go) in relation to the second visual cues. Some of the visual category-specific neurons in the CMAr further displayed the motor response-specific activity. On the other hand, many of the task-related CMAd and CMAv neurons seemed to be implicated directly in motor functions, such as preparation and execution of movements in Go trials. The present results suggest that the CMAr neurons may participate in cognitive and motor functions of “attention for action” and “response selection” for an appropriate action according to an intention, whereas the CMAd and CMAv neurons may be involved in “motor preparation and execution”.