Individual differences in sensory responses influence decision making by Drosophila melanogaster larvae on exposure to contradictory cues.

Animals make decisions on behavioral choice by evaluating internal and external signals. Individuals often make decisions in different ways, but the underlying neural mechanisms are not well understood. Here, we describe a system for observing the behavior of individual Drosophila melanogaster larvae simultaneously presented with contradictory signals, in this case attractive (yeast paste) and aversive (NaCl) signals. Olfaction was used to detect the yeast paste, whereas the ENaC/Pickpocket channel was important for NaCl detection. We found that wild-type (Canton-S) larvae fall into two decision making groups: one group decided to approach the yeast paste by overcoming the aversive signal, whereas the other group decided to forgo the yeast paste because of the aversive signal. Our findings indicate that different endogenous sensitivities to NaCl contribute to make differences between two groups and that diverse decision making steps occur in individual animals.

Intracellular calcium elevation during plateau potentials mediated by extrasynaptic NMDA receptor activation in rat hippocampal CA1 pyramidal neurons is primarily due to calcium entry through voltage-gated calcium channels.

We reported previously that plateau potentials mediated by extrasynaptic N-methyl-d-aspartate receptors (NMDARs) can be induced either by synaptic stimulation in the presence of glutamate transporter antagonist or by iontophoresis of NMDA in rat hippocampal CA1 pyramidal neurons. To examine whether the plateau potentials are accompanied by an elevation of intracellular Ca2+ and to determine the source of Ca2+ elevation, we performed Ca2+ imaging during the plateau potential. Neurons were loaded with Ca2+ indicator fluo-4, and the plateau potentials were generated either synaptically in the presence of glutamate transporter antagonist or by iontophoretically applying NMDA. We have found that a transient elevation in intracellular Ca2+ accompanies the plateau potential. The synaptically induced plateau potential and the Ca2+ elevation were blocked by 5,7-dichlorokynurenic acid (5,7-dCK), an antagonist for the glycine-binding sites of NMDAR. A mixture of Cd2+ and tetrodotoxin did not block NMDA-induced plateau potentials, but completely abolished the accompanying Ca2+ elevation in both the presence and absence of Mg2+ ions in the bathing solution. The NMDA-induced plateau potential was blocked by further adding 5,7-dCK. Our results show that the NMDAR-mediated plateau potential is accompanied by elevation of intracellular Ca2+ that is primarily caused by the influx of Ca2+ through voltage-gated Ca2+ channels.

Downregulation of Centaurin gamma1A increases synaptic transmission at Drosophila larval neuromuscular junctions.

Adequate regulation of synaptic transmission is critical for appropriate neural circuit functioning. Although a number of molecules involved in synaptic neurotransmission have been identified, the molecular mechanisms regulating neurotransmission are not fully understood. Here, we focused on Centaurin gamma1A (CenG1A) and examined its role in synaptic transmission regulation using Drosophila larval neuromuscular junctions. CenG1A is a member of the Centaurin family, which contains Pleckstrin homology, ADP ribosylation factor GTPase-activating protein, and ankyrin repeat domains. Due to the existence of these functional domains, CenG1A is proposed to be involved in the process of synaptic release; however, no evidence for this has been found to date. In this study, we investigated the potential role for CenG1A in the process of synaptic release by performing intracellular recordings in larval muscle cells. We found that neurotransmitter release from presynaptic cells was enhanced in cenG1A mutants. This effect was also observed in larvae with reduced CenG1A function in either presynaptic or postsynaptic cells. In addition, we revealed that suppressing CenG1A function in postsynaptic muscle cells led to an increase in the probability of neurotransmitter release, whereas its suppression in presynaptic neurons led to an increase in neurotransmitter release probability and an increase in the number of synaptic vesicles. These results suggested that CenG1A functions at both presynaptic and postsynaptic sites as a negative regulator of neurotransmitter release. Our study provided evidence for a key role of CenG1A in proper synaptic transmission at neuromuscular junctions.

Pitfalls in the dipolar model for the neocortical EEG sources.

For about six decades, primary current sources of the electroencephalogram (EEG) have been assumed dipolar in nature. In this study, we used electrophysiological recordings from anesthetized Wistar rats undergoing repeated whisker deflections to revise the biophysical foundations of the EEG dipolar model. In a first experiment, we performed three-dimensional recordings of extracellular potentials from a large portion of the barrel field to estimate intracortical multipolar moments generated either by single spiking neurons (i.e., pyramidal cells, PC; spiny stellate cells, SS) or by populations of them while experiencing synchronized postsynaptic potentials. As expected, backpropagating spikes along PC dendrites caused dipolar field components larger in the direction perpendicular to the cortical surface (49.7 ± 22.0 nA·mm). In agreement with the fact that SS cells have "close-field" configurations, their dipolar moment at any direction was negligible. Surprisingly, monopolar field components were detectable both at the level of single units (i.e., -11.7 ± 3.4 nA for PC) and at the mesoscopic level of mixed neuronal populations receiving extended synaptic inputs within either a cortical column (-0.44 ± 0.20 µA) or a 2.5-m(3)-voxel volume (-3.32 ± 1.20 µA). To evaluate the relationship between the macroscopically defined EEG equivalent dipole and the mesoscopic intracortical multipolar moments, we performed concurrent recordings of high-resolution skull EEG and laminar local field potentials. From this second experiment, we estimated the time-varying EEG equivalent dipole for the entire barrel field using either a multiple dipole fitting or a distributed type of EEG inverse solution. We demonstrated that mesoscopic multipolar components are altogether absorbed by any equivalent dipole in both types of inverse solutions. We conclude that the primary current sources of the EEG in the neocortex of rodents are not precisely represented by a single equivalent dipole and that the existence of monopolar components must be also considered at the mesoscopic level.

Experience-dependent plasticity of the optomotor response in Drosophila melanogaster.

Experience in early life can affect the development of the nervous system. There is now evidence that experience-dependent plasticity exists in adult insects. To uncover the molecular basis of plasticity, an invertebrate model, such as Drosophila melanogaster, is a powerful tool, as many established genetic and molecular methods can be applied. To establish a model system in which behavioral plasticity can be examined, we investigated the optomotor response, a behavior common to most sight-reliant animals, in Drosophila and found that the response could be modified by the level of light during rearing. The angle turned by the head in response to a moving stimulus was used to quantify the response. Deprivation of light increased the response to low-contrast stimuli in wild-type Drosophila at 4 days after eclosion and this plastic change did not appear in rutabaga, a known mutant defective in short-term memory. In addition, the change was transient and was markedly decreased at 6 days after eclosion. Further, we found that Dark-flies, which have been kept in constant darkness for more than 50 years, showed a higher response to low-contrast stimuli even at 6 days after eclosion compared to wild type and this characteristic was not lost in Dark-flies placed in a normal light environment for 2 generations, suggesting that this high response has a hereditary nature. Thus, our model system can be used to examine how the environment affects behaviors.

Imaging data analysis using non-negative matrix factorization.

The rapid progress of imaging devices such as two-photon microscopes has made it possible to measure the activity of thousands to tens of thousands of cells at single-cell resolution in a wide field of view (FOV) data. However, it is not possible to manually identify thousands of cells in such wide FOV data. Several research groups have developed machine learning methods for automatically detecting cells from wide FOV data. Many of the recently proposed methods using dynamic activity information rather than static morphological information are based on non-negative matrix factorization (NMF). In this review, we outline cell-detection methods related to NMF. For the purpose of raising issues on NMF cell detection, we introduce our current development of a non-NMF method that is capable of detecting about 17,000 cells in ultra-wide FOV data.

An analytic solution of the cable equation predicts frequency preference of a passive shunt-end cylindrical cable in response to extracellular oscillating electric fields.

Under physiological and artificial conditions, the dendrites of neurons can be exposed to electric fields. Recent experimental studies suggested that the membrane resistivity of the distal apical dendrites of cortical and hippocampal pyramidal neurons may be significantly lower than that of the proximal dendrites and the soma. To understand the behavior of dendrites in time-varying extracellular electric fields, we analytically solved cable equations for finite cylindrical cables with and without a leak conductance attached to one end by employing the Green's function method. The solution for a cable with a leak at one end for direct-current step electric fields shows a reversal in polarization at the leaky end, as has been previously shown by employing the separation of variables method and Fourier series expansion. The solution for a cable with a leak at one end for alternating-current electric fields reveals that the leaky end shows frequency preference in the response amplitude. Our results predict that a passive dendrite with low resistivity at the distal end would show frequency preference in response to sinusoidal extracellular local field potentials. The Green's function obtained in our study can be used to calculate response for any extracellular electric field.

Selection of behaviors and segmental coordination during larval locomotion is disrupted by nuclear polyglutamine inclusions in a new Drosophila Huntington's disease-like model.

Huntington's disease is an autosomal dominant neurodegenerative disorder that is caused by abnormal expansion of a polyglutamine tract in the huntingtin protein, resulting in intracellular aggregate formation and neurodegeneration. How neuronal cells are affected by such a polyglutamine tract expansion remains obscure. To dissect the ways in which polyglutamine expansion can cause neural dysfunction, the authors generated Drosophila transgenic strains expressing either a nuclear targeted or cytoplasmic form of pathogenic (NHtt-152Q(NLS), NHtt-152Q), or nonpathogenic (NHtt-18Q(NLS), NHtt-18Q) N-terminal human huntingtin. These proteins were expressed in the dendritic arborization neurons of the larval peripheral nervous system and their effects on neuronal survival, morphology, and larval locomotion were examined. The authors found that NHtt-152Q(NLS) larvae had altered dendrite morphology and larval locomotion, whereas NHtt-152Q, NHtt-18Q(NLS), and NHtt-18Q larvae did not. Furthermore, the authors examined the physiological defect underlying this disrupted larval locomotion in detail by recording spontaneous ongoing segmental nerve activity. NHtt-152Q(NLS) larvae displayed uncoordinated activity between anterior and posterior segments. Moreover, anterior segments had shorter bursts and longer interburst intervals in NHtt-152Q(NLS) larvae than in NHtt-18Q(NLS) larvae, whereas posterior segments had longer bursts and shorter interburst intervals. These results suggest that the pathogenic protein disrupts neuron function without inducing cell death, and describe how this dysfunction leads to a locomotor defect. These results also suggest that sensory inputs are necessary for the coordination of anterior and posterior body parts during locomotion. From these analyses the authors show that examination of motor behaviors in the Drosophila larvae is a powerful new model to dissect non-cell-lethal mechanisms of mutant Htt toxicity.

Frequency-dependent entrainment of spontaneous Ca transients in the dendritic tufts of CA1 pyramidal cells in rat hippocampal slice preparations by weak AC electric field.

Neurons in the central nervous systems are exposed to endogenous oscillating electric fields and their activities are likely to be modified by those fields. We had previously investigated the effects of AC electric field by using a newly developed method to monitor local Ca transients in the dendrites of a neuronal population in acute rat hippocampal slices and reported that spontaneously occurring Ca transients in the tufts of the apical dendrites of CA1 pyramidal neurons become entrained to subthreshold AC electric fields. To further our understanding of the impact of AC fields on dendritic activities, in the present study we examined three questions: how does the extent of entrainment depend on the frequency of the applied field, how does the mean phase of the dendritic activities during field application depend on the frequency of the field, and whether the entrainment can be seen in the absence of synaptic transmission. We have found that, the extent of entrainment is significantly greater at a low frequency band (1-4 Hz) compared to a high frequency band (8-16 Hz), 0.688 ± 0.027 at 2 Hz compared to 0.087 ± 0.016 at 16 Hz in case of 7 mV/mm field strength, that the entrainment can be observed when synaptic transmission is pharmacologically blocked, and that the mean phase of the Ca transients during field stimulation at a low frequency band (1-4 Hz) stays constant. These results indicate that the electric fields with physiologically feasible frequencies and intensities can entrain activities of the dendrites in a frequency-dependent manner independent of synaptic transmission. AC electric fields during oscillatory brain activities might play a role in synchronizing neural activities by modulating dendritic activities.

A plateau potential mediated by the activation of extrasynaptic NMDA receptors in rat hippocampal CA1 pyramidal neurons.

Hippocampal pyramidal neurons express various extrasynaptic glutamate receptors. When glutamate spillover was facilitated by blocking glutamate uptake and fast synaptic transmission was blocked by antagonists of AMPA- and NMDA-type glutamate receptors and an ionotropic GABA receptor blocker, repetitive synaptic stimulation evoked a persistent membrane depolarization that consisted of an early Ca(2+)-independent component and a late Ca(2+)-dependent component. The early component, which we refer to as a plateau potential, had a half-width of 770 +/- 160 ms and a steady peak level of -9.54 +/- 3.50 mV. It was accompanied by an increase in membrane conductance, the I-V relationship of which showed a peak at -19.91 +/- 2.18 mV and reversal of the current at -4.32 +/- 2.13 mV, and was suppressed by high concentration of an NMDA receptor (NMDAR) antagonist d-APV, or an NMDAR glycine-binding site antagonist 5,7-dCK. After blocking synaptically located NMDARs using MK801, the potential was still evoked synaptically when spillover was facilitated. A sustained depolarization was evoked by iontophoretic application of glutamate in the presence or absence of a glutamate uptake blocker. This potential was not affected by Na(+) or Ca(2+) channel blockers, but was suppressed by 5,7-dCK, leaving an unspecified depolarizing potential. Iontophoresis of NMDA evoked a sustained depolarization that was blocked by a high concentration of d-APV or 5,7-dCK. The I-V relationship of the current during this potential was similar to that obtained during the synaptically induced plateau potentials. These results show that CA1 pyramidal neurons generate plateau potentials mediated most likely by activation of extrasynaptic NMDARs.

Involvement of basal protein kinase C and extracellular signal-regulated kinase 1/2 activities in constitutive internalization of AMPA receptors in cerebellar Purkinje cells.

AMPA receptor (AMPAR) internalization provides a mechanism for long-term depression (LTD) in both hippocampal pyramidal neurons and cerebellar Purkinje cells (PCs). Cerebellar LTD at the parallel fiber (PF)-PC synapse is the underlying basis of motor learning and requires AMPAR activation, a large Ca2+ influx, and protein kinase C (PKC) activation. However, whether these requirements affect the constitutive AMPAR internalization in PF-PC synapses remains unclarified. Tetanus toxin (TeTx) infusion into PCs decreased PF-EPSC amplitude to 60% within 20-30 min (TeTx rundown), without change in paired-pulse facilitation ratio or receptor kinetics. Immunocytochemically measured glutamate receptor 2 (GluR2) internalization ratio decreased at the steady state of TeTx rundown. TeTx rundown did not require AMPAR activity nor an increase in intracellular Ca2+ concentration. TeTx rundown was suppressed partially by the inhibition of either conventional PKC or mitogen-activated protein kinase kinase (MEK) and completely by the inhibition of both kinases. The background PKC activity was shown to be sufficient, because a PKC activator did not facilitate TeTx rundown. The inhibition of protein phosphatase 1/2A (PP1/2A) enhanced TeTx rundown slightly, and both inhibition of PP1/2A and activation of PKC maximized it, but one-half of AMPARs at PF-PC synapses remained in the TeTx-resistant pool. The inhibition of actin depolymerization suppressed TeTx rundown and decreased the GluR2 internalization ratio. In contrast, the inhibition of actin polymerization enhanced TeTx rundown and increased the GluR2 internalization ratio. We suggest that the regulation of actin polymerization is involved in the surface expression of AMPARs and the surface expressing AMPARs are constitutively internalized through both basal PKC and MEK-ERK1/2 (extracellular signal-regulated kinase 1/2) activities at PF-PC synapses.

Paired-pulse ratio of synaptically induced transporter currents at hippocampal CA1 synapses is not related to release probability.

When a synapse is stimulated in rapid succession, the second post-synaptic response can be larger than the first and termed paired-pulse facilitation. It has been reported that the paired-pulse ratio (PPR), which is the ratio of the amplitude of the second response to that of the first, depends on the probability of vesicular release at the synapse, and PPR has been used as an easy measure of the release probability. To re-examine the relation of PPR with transmitter release probability, we made whole-cell recordings from astrocytes and pyramidal neurons in the CA1 area of rat hippocampal slices, and studied responses evoked by paired-pulse stimulus of the Schaffer collaterals. In a control condition in which blockers for ionotropic glutamate receptors were added to the artificial cerebrospinal fluid, synaptically induced transporter currents (STCs) recorded from astrocytes showed PPF with similar dependency on stimulus interval as the AMPA-receptor-mediated excitatory post-synaptic currents (AMPA-EPSCs) recorded from pyramidal neurons. When the transmitter release was enhanced by raising Ca2+ concentration in the bathing medium or by applying 8-CPT, an adenosine A1 receptor antagonist, the PPR of the neuronal AMPA-EPSCs decreased significantly. In the same condition, although the amplitude of STCs was significantly increased, the PPR of STCs did not show significant change. The PPR of AMPA-EPSCs, however, recovered by lowering the stimulus intensity or by applying low concentration of NBQX, a competitive antagonist for AMPA-receptor. These results imply that the PPR of transmitter release at Schaffer collateral synapses stays constant as the release probability was altered.

Dendritic Spikes in Sensory Perception.

What is the function of dendritic spikes? One might argue that they provide conditions for neuronal plasticity or that they are essential for neural computation. However, despite a long history of dendritic research, the physiological relevance of dendritic spikes in brain function remains unknown. This could stem from the fact that most studies on dendrites have been performed in vitro. Fortunately, the emergence of novel techniques such as improved two-photon microscopy, genetically encoded calcium indicators (GECIs), and optogenetic tools has provided the means for vital breakthroughs in in vivo dendritic research. These technologies enable the investigation of the functions of dendritic spikes in behaving animals, and thus, help uncover the causal relationship between dendritic spikes, and sensory information processing and synaptic plasticity. Understanding the roles of dendritic spikes in brain function would provide mechanistic insight into the relationship between the brain and the mind. In this review article, we summarize the results of studies on dendritic spikes from a historical perspective and discuss the recent advances in our understanding of the role of dendritic spikes in sensory perception.

Estimated distribution of specific membrane resistance in hippocampal CA1 pyramidal neuron.

It has been suggested that dendritic membrane properties play an important role in a synaptic integration. In particular, the specific membrane resistance, one of membrane properties, has been reported to be non-uniformly distributed in a single neuron, although the spatial distribution of the specific membrane resistance is still unclear. To reveal its non-uniformity in dendrite, we estimated the spatial distribution of specific membrane resistance in a single neuron, based on voltage imaging data, observed optically in hippocampal CA1 slices. As the optically recorded data, we used bi-directional propagations of subthreshold excitatory postsynaptic potentials in dendrite, which were not be reproduced numerically with uniform-specific membrane resistance. By numerical simulations for multi-compartment models with non-uniformity of specific membrane resistance, we estimated that the distribution obeys a step function; the optically recorded data were consistently reproduced for the distribution with a steep decrease in the specific membrane resistance at the distal apical dendrite, which occurs 300-500 microm away from the soma. In the estimated distribution, the specific membrane resistance at the distal side is less than about 10(3) Omegacm(2), whereas the resistance at the proximal side is greater than about 10(4) Omegacm(2). This result implies that the specific membrane resistance decreases drastically at the distal apical dendrite in hippocampal CA1 pyramidal neuron.

Organization of projection neurons and local neurons of the primary auditory center in the fruit fly Drosophila melanogaster.

Acoustic communication between insects serves as an excellent model system for analyzing the neuronal mechanisms underlying auditory information processing. The detailed organization of auditory neural circuits in the brain has not yet been described. To understand the central auditory pathways, we used the brain of the fruit fly Drosophila melanogaster as a model and performed a large-scale analysis of the interneurons associated with the primary auditory center. By screening expression driver strains and performing single-cell labeling of these strains, we identified 44 types of interneurons innervating the primary auditory center. Five types were local interneurons whereas the other 39 types were projection interneurons connecting the primary auditory center with other brain regions. The projection neurons comprised three frequency-selective pathways and two frequency-embracive pathways. Mapping of their connection targets revealed that five neuropils in the brain-the wedge (WED), anterior ventrolateral protocerebrum, posterior ventrolateral protocerebrum (PVLP), saddle (SAD), and gnathal ganglia (GNG)-were intensively connected with the primary auditory center. In addition, several other neuropils, including visual and olfactory centers in the brain, were directly connected to the primary auditory center. The distribution patterns of the spines and boutons of the identified neurons suggest that auditory information is sent mainly from the primary auditory center to the PVLP, WED, SAD, GNG, and thoracico-abdominal ganglia. Based on these findings, we established the first comprehensive map of secondary auditory interneurons, which indicates the downstream information flow to parallel ascending pathways, multimodal pathways, and descending pathways.

Weak sinusoidal electric fields entrain spontaneous Ca transients in the dendritic tufts of CA1 pyramidal cells in rat hippocampal slice preparations.

Neurons might interact via electric fields and this notion has been referred to as ephaptic interaction. It has been shown that various types of ion channels are distributed along the dendrites and are capable of supporting generation of dendritic spikes. We hypothesized that generation of dendritic spikes play important roles in the ephaptic interactions either by amplifying the impact of electric fields or by providing current source to generate electric fields. To test if dendritic activities can be modulated by electric fields, we developed a method to monitor local Ca-transients in the dendrites of a neuronal population in acute rat hippocampal slices by applying spinning-disk confocal microscopy and multi-cell dye loading technique. In a condition in which the dendrites of CA1 pyramidal neurons show spontaneous Ca-transients due to added 50 µM 4-aminopyridine to the bathing medium and adjusted extracellular potassium concentration, we examined the impact of sinusoidal electric fields on the Ca-transients. We have found that spontaneously occurring fast-Ca-transients in the tufts of the apical dendrites of CA1 pyramidal neurons can be blocked by applying 1 µM tetrodotoxin, and that the timing of the transients become entrained to sub-threshold 1-4 Hz electric fields with an intensity as weak as 0.84 mV/mm applied parallel to the somato-dendritic axis of the neurons. The extent of entrainment increases with intensity below 5 mV/mm, but does not increase further over the range of 5-20 mV/mm. These results suggest that population of pyramidal cells might be able to detect electric fields with biologically relevant intensity by modulating the timing of dendritic spikes.

Noise-robust recognition of wide-field motion direction and the underlying neural mechanisms in Drosophila melanogaster.

Appropriate and robust behavioral control in a noisy environment is important for the survival of most organisms. Understanding such robust behavioral control has been an attractive subject in neuroscience research. Here, we investigated the processing of wide-field motion with random dot noise at both the behavioral and neuronal level in Drosophila melanogaster. We measured the head yaw optomotor response (OMR) and the activity of motion-sensitive neurons, horizontal system (HS) cells, with in vivo whole-cell patch clamp recordings at various levels of noise intensity. We found that flies had a robust sensation of motion direction under noisy conditions, while membrane potential changes of HS cells were not correlated with behavioral responses. By applying signal classification theory to the distributions of HS cell responses, however, we found that motion direction under noise can be clearly discriminated by HS cells, and that this discrimination performance was quantitatively similar to that of OMR. Furthermore, we successfully reproduced HS cell activity in response to noisy motion stimuli with a local motion detector model including a spatial filter and threshold function. This study provides evidence for the physiological basis of noise-robust behavior in a tiny insect brain.

Arthritis induced in cat by sodium urate: a possible animal model for tonic pain.

An attempt has been made to determine whether cats rendered arthritic by the injection of monosodium urate (MSU) crystals (rod-shaped 40-130 micrometers length) into one knee joint capsule can be used as animal model of tonic (chronic) pain. A limp and a decrease in body weight supported by the injected hind leg's paw occurred approximately 1 h after the MSU (20 mg) injection, reached a maximum at 2-3 h, and lasted for more than 6 h before a gradual return to pre-injection levels. They were diminished by systemic administration and local (the dorsal part of the nucleus raphe dorsalis) application of morphine, this effect being blocked by naloxone. This suggests that the limping and the paw pressure decrease are the reflexion of pain. It is suggested that the animal model of the MSU-induced arthritis is useful for the study of tonic pain.

NMDA receptor-mediated depolarizing after-potentials in the basal dendrites of CA1 pyramidal neurons.

It was shown recently that the basal dendrites of cortical pyramidal neurons generate NMDA-spikes. In the present study, we made whole-cell recordings from hippocampal CA1 pyramidal neurons and examined whether NMDA receptor activation was involved in synaptic responses. At low input stimulus intensity, EPSPs with a fast decay time were induced. As the intensity of stimulation was increased in the presence of GABA receptor antagonists, a depolarizing after-potential (DAP) was generated in addition to a fast decaying potential. A DAP was never observed when the input was applied to the apical dendrites. The DAP was suppressed by hyperpolarization or by NMDA receptor antagonists, but not by Na+, K+, or Ca2+ channel blockers. One possible mechanism is that the morphology of the basal dendrites favors DAP generation. A compartmental model simulation showed that synaptic inputs to thinner shorter dendrites generated a potential that resembled a DAP. Our study shows that a synaptic input to the basal dendrites of a hippocampal pyramidal neuron can generate a NMDA receptor-mediated potential in the presence of GABA receptor blockade.

Adenosine A(1)-receptor-mediated tonic inhibition of glutamate release at rat hippocampal CA3-CA1 synapses is primarily due to inhibition of N-type Ca(2+) channels.

The voltage-gated Ca(2+) channels responsible for synaptic transmission at CA3-CA1 synapses are mainly P/Q- and N-types. It has been shown that tonic inhibition of transmission due to activation of adenosine A(1) receptors occurs at this synapse. We have recently developed a technique to monitor synaptically released glutamate which is based on synaptically induced glial depolarisation. Using this technique, we have examined the effects of different voltage-gated Ca(2+) channel blockers on glutamate release. Under conditions in which the adenosine A(1) receptor was not blocked, omega-AgaIVA (a P/Q-type voltage-gated Ca(2+) channel blocker) suppressed synaptically induced glial depolarisation to a greater extent than omega-CgTxGVIA (an N-type voltage-gated Ca(2+) channel blocker) did. In contrast, in the presence of an adenosine A(1) receptor antagonist, omega-AgaIVA was less effective at suppressing synaptically induced glial depolarisation than omega-CgTxGVIA. These results indicate that, in the absence of adenosine A(1) receptor-mediated tonic inhibition, the contribution of N-type is much greater than that of P-type, and that N-types are the primary target of tonic inhibition in normal conditions in which adenosine A(1) receptor-mediated tonic inhibition is present.