Intracellular injection of EGTA abolished the hyperpolarization seen after spontaneous and evoked... more Intracellular injection of EGTA abolished the hyperpolarization seen after spontaneous and evoked bursts of action potentials in hippocampal CA3 neurons. Simultaneously recorded, orthodromically elicited IPSPs were relatively unaffected by this treatment. It is concluded that CA3 neurons generate at least two types of hyperpolarization: one resulting from a calcium-mediated potassium conductance and the other from increased conductance to chloride.
Quisqualate induces an inward current via mGluR activation in neocortical pyramidal neurons
Brain Research, 2000
Activation of metabotropic glutamate receptors (mGluRs) has multiple effects on the excitability ... more Activation of metabotropic glutamate receptors (mGluRs) has multiple effects on the excitability of pyramidal neurons in rat frontal neocortex. Synaptic transmission and intrinsic excitability are both affected. During studies of the effects of quisqualate on synaptic activity, it was observed that quisqualate also induced a slow inward current. Whole-cell patch clamp recordings were obtained from layer II/III pyramidal neurons of neocortical slices in vitro. The bath solution contained APV, CNQX and bicuculline to block ionotropic glutamate and GABA(A) receptors. At a holding potential of -70 mV, quisqualate (2 microM) induced an inward current of about 60 pA. The response was reversible upon washing. This current was associated with an increase in membrane conductance and was still seen in the presence of TTX (0.5 microM). Bath application of the nonselective mGluR antagonist, (R, S)-alpha-methyl-4-carboxyphenyglycine (MCPG, 200-500 microM) reduced the current by 70%. Other mGluR agonists (ACPD, DHPG, L-CCG-1 and L-AP4) did not induce a significant inward current at the concentrations tested. The current-voltage relation of the quisqualate-induced current was linear with a reversal potential near 0 mV suggesting involvement of nonselective cation channels. The quisqualate-induced inward current was markedly reduced (72%) with 200 microM GDP-beta-S in the pipette solution, indicating that it is a postsynaptic phenomenon mediated by a G-protein dependent mechanism. These results suggest that mGluRs can directly increase the postsynaptic excitability of pyramidal cells.
Cultures of cerebral neurons were prepared from chick embryos, 8.5 days in ovo, and maintained in... more Cultures of cerebral neurons were prepared from chick embryos, 8.5 days in ovo, and maintained in vitro. Following chronic exposure of these cells to GABA, the levels of [ 3 H]flunitrazepam binding in situ and electrophysiological responsiveness to γ-aminobutyric acid (GABA) was ...
Zinc enhances GABAergic transmission in rat neocortical neurons
Journal of Neurophysiology
1. Intracellular recordings were made in layer II-III neurons of rat neocortical slices maintaine... more 1. Intracellular recordings were made in layer II-III neurons of rat neocortical slices maintained in vitro. The effect of bath application of zinc (50-300 microM) on evoked synaptic activity and passive membrane properties was examined. 2. Excitatory postsynaptic potentials (EPSPs) mediated by N-methyl-D-aspartate (NMDA) and non-NMDA receptors were recorded in response to electrical stimulation. Zinc did not affect either type of EPSP. Resting membrane potential, repetitive firing properties, and input resistance were not altered by zinc. 3. Inhibitory postsynaptic potentials (IPSPs) were enhanced after zinc application. Zinc also induced generation of large amplitude spontaneous gamma-aminobutyric acid-A (GABAA)- and GABAB-mediated IPSPs. Postsynaptic responses to iontophoretically applied GABA were unaffected. In the presence of zinc, GABAergic synaptic potentials could result in generation of action potentials. 4. Directly evoked IPSPs recorded in the presence of the excitatory ...
Layer I neurons of the rat neocortex. II. Voltage-dependent outward currents
Journal of Neurophysiology
1. Whole cell patch-clamp techniques, combined with direct visualization of neurons, were used to... more 1. Whole cell patch-clamp techniques, combined with direct visualization of neurons, were used to study voltage-dependent potassium currents in layer 1 neurons and layer II/III pyramidal cells. 2. In the presence of tetrodotoxin, step depolarizations evoked an outward current. This current had a complex waveform and appeared to be a composite of early and late components. The early peak of the composite K+ outward current was larger in layer I neurons. 3. In both layer I and pyramidal cells, the composite outward K+ current could be separated into two components based on kinetic and pharmacological properties. The early component was termed I(A) because it was a transient outward current activating rapidly and then decaying. I(A) was more sensitive to blocking by 4-aminopyridine (4-AP) than tetraethylammonium (TEA). The second component, termed the delayed rectifier or I(DR), activated relatively slowly and did not decay significantly during a 200-ms test pulse. I(DR) was insensitiv...
The Journal of neuroscience : the official journal of the Society for Neuroscience, Jan 18, 2015
Circuit dysfunction in complex brain disorders such as schizophrenia and autism is caused by imba... more Circuit dysfunction in complex brain disorders such as schizophrenia and autism is caused by imbalances between inhibitory and excitatory synaptic transmission (I/E). Short-term plasticity differentially alters responses from excitatory and inhibitory synapses, causing the I/E ratio to change as a function of frequency. However, little is known about I/E ratio dynamics in complex brain disorders. Transcriptional dysregulation in interneurons, particularly parvalbumin interneurons, is a consistent pathophysiological feature of schizophrenia. Peroxisome proliferator activated receptor γ coactivator 1α (PGC-1α) is a transcriptional coactivator that in hippocampus is highly concentrated in inhibitory interneurons and regulates parvalbumin transcription. Here, we used PGC-1α(-/-) mice to investigate effects of interneuron transcriptional dysregulation on the dynamics of the I/E ratio at the synaptic and circuit level in hippocampus. We find that loss of PGC-1α increases the I/E ratio ont...
Enhanced receptiveness at all synapses on a neuron that receive glutamatergic input is called cel... more Enhanced receptiveness at all synapses on a neuron that receive glutamatergic input is called cell-wide synaptic upscaling. We hypothesize that this type of synaptic plasticity may be critical for long-term memory storage within cortical circuits, a process that may also depend on epigenetic mechanisms, such as covalent chemical modification of DNA. We found that DNA cytosine demethylation mediates multiplicative synaptic upscaling of glutamatergic synaptic strength in cultured cortical neurons. Inhibiting neuronal activity with tetrodotoxin (TTX) decreased the cytosine methylation of and increased the expression of genes encoding glutamate receptors and trafficking proteins, in turn increasing the amplitude but not frequency of miniature excitatory postsynaptic currents (mEPSCs), indicating synaptic upscaling rather than increased spontaneous activity. Inhibiting DNA methyltransferase (DNMT) activity, either by using the small-molecule inhibitor RG108 or by knocking down Dnmt1 and ...
The cerebral cortex receives an extensive serotonergic (5-hydroxytryptamine, 5-HT) input. Immunoh... more The cerebral cortex receives an extensive serotonergic (5-hydroxytryptamine, 5-HT) input. Immunohistochemical studies suggest that inhibitory neurons are the main target of 5-HT innervation. In vivo extracellular recordings have shown that 5-HT generally inhibited cortical pyramidal neurons, whereas in vitro studies have shown an excitatory action. To determine the cellular mechanisms underlying the diverse actions of 5-HT in the cortex, we examined its effects on cortical inhibitory interneurons and pyramidal neurons. We found that 5-HT, through activation of 5-HT(2A) receptors, induced a massive enhancement of spontaneous inhibitory postsynaptic currents (sIPSCs) in pyramidal neurons, lasting for approximately 6 min. In interneurons, this 5-HT-induced enhancement of sIPSCs was much weaker. Activation of 5-HT(2A) receptors also increased spontaneous excitatory postsynaptic currents (sEPSCs) in pyramidal neurons. This response desensitized less and at a slower rate. In contrast, 5-H...
Early postnatal freeze lesions in rat neocortex produce anatomic abnormalities resembling those o... more Early postnatal freeze lesions in rat neocortex produce anatomic abnormalities resembling those observed in human patients with seizure disorders. Although in vitro brain slices containing the lesion are hyperexcitable, the mechanisms of this alteration have yet to be elucidated. To test the hypothesis that changes in postsynaptic inhibitory receptors may underlie this hyperexcitability, we examined properties of gamma-aminobutyric acid type A receptor (GABAAR)-mediated miniature inhibitory postsynaptic currents (mIPSCs). Recordings were obtained in layer II/III pyramidal cells located 1-2 mm lateral to the lesion. mIPSC peak amplitude and rate of rise were increased relative to nonlesioned animals, whereas decay time constant and interevent interval were unaltered. Bath application of zolpidem at a concentration (20 nM) specific for activation of the type 1 benzodiazepine receptor had no significant effect on decay time constant in six of nine cells. Exposure to higher concentratio...
The effect of metabotropic glutamate receptor (mGluR) activation on inhibitory synaptic transmiss... more The effect of metabotropic glutamate receptor (mGluR) activation on inhibitory synaptic transmission was examined by using whole cell patch-clamp recordings. Spontaneous (s) and miniature (m) inhibitory postsynaptic currents (IPSCs) were recorded from visually identified layer II/III pyramidal neurons in rat neocortex in vitro. Excitatory postsynaptic currents (EPSCs) were blocked by using bath application of 20 microM D(-)2-amino-5-phosphonovaleric acid and 10 microM 6-cyano-7-nitroquinoxaline-2,3-dione. In the presence of 1S,3R-1-aminocyclopentane-1,3-dicarboxylic acid (30-100 microM), Lp4-quisqualate (5 microM), and the group I selective mGluR agonist (S)-3,5-dihydroxyphenylglycine (100 microM), the frequency of sIPSCs was increased. Decay kinetics of sIPSCs were unaffected. No enhancement of mIPSCs was observed. Bath application of group II (2S,3S,4S-alpha-carboxycyclopropyl-glycine; 5 microM) and group III selective mGluR agonists (L-2-amino-4-phosphonobutyric acid; 100 microM)...
With the use of the whole cell patch-clamp technique combined with visualization of neurons in br... more With the use of the whole cell patch-clamp technique combined with visualization of neurons in brain slices, we studied the properties of miniature excitatory postsynaptic currents (mEPSCs) in rat neocortical layer I neurons. At holding potentials (-50 to -70 mV) near the resting membrane potential (RMP), mEPSCs had amplitudes of 5-100 pA and were mediated mostly by alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionate (AMPA) receptors. Amplitude histograms were skewed toward large events. An N-methyl-D-aspartate (NMDA) component was revealed by depolarization to -30 mV or by the use of a Mg2+-free bathing solution. At RMP, averaged AMPA mEPSCs had a 10-90% rise time of approximately 0.3 ms (uncorrected for instrument filtering). The decay of averaged mEPSCs was best fit by double-exponential functions in most cases. The fast, dominating component had a decay time constant of approximately 1.2 ms and comprised approximately 80% of the total amplitude. A small slow component had a de...
1. Whole-cell patch clamp techniques were used to record transient outward potassium currents in ... more 1. Whole-cell patch clamp techniques were used to record transient outward potassium currents in embryonic rat neocortical neurons maintained in culture. The effect of lidocaine and its quaternary derivative QX222 on this transient outward current (TOC) was examined. 2. Extracellular application of lidocaine produced a progressive decrease in peak TOC amplitude with no change in the overall current waveform. Peak amplitude was reduced to 74 +/- 6% (mean +/- SE) of control by 0.5 mM and to 48 +/- 2% by 2 mM lidocaine. The effect of lidocaine was fully reversible. Intracellular application of QX222 also resulted in a concentration-dependent reduction of the TOC with 0.5 mM reducing the peak amplitude to 73 +/- 8% of control and 1 mM reducing it to 41 +/- 4%. 3. Lidocaine reduced TOCs when applied intracellularly but the rate of block was considerably slower than with extracellular application. QX222 had no effect on the TOC when applied extracellularly. 4. Lidocaine (1 mM) induced a 4...
Developmental changes in NMDA and non-NMDA receptor-mediated synaptic potentials in rat neocortex
Journal of neurophysiology, 1993
1. In vitro slices of frontal neocortex were prepared from rat pups 3-14 days of age. Whole-cell ... more 1. In vitro slices of frontal neocortex were prepared from rat pups 3-14 days of age. Whole-cell patch-clamp recordings were obtained from layer II-III cortical neurons, and measurements of passive membrane properties were made. The development of evoked synaptic excitation and inhibition was also examined with the use of current- and voltage-clamp techniques. 2. Pharmacological separation of excitatory synaptic activity into both N-methyl-D-aspartate (NMDA) and non-NMDA receptor-mediated components was accomplished by application of D(-)2-amino-5-phosphonovaleric acid (APV), D(-)2-amino-7-phosphonoheptanoic acid (AP7), and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). Inhibitory synaptic events were described according to their reversal potentials and modulation by the GABAA receptor antagonist bicuculline methiodide (BMI). 3. Pups were grouped into three categories on the basis of age: postnatal day (PN) 3-5, PN 6-8, and PN 9-14. In slices from PN 3-5 pups, neurons exhibited high i...
NMDA receptor-mediated components of miniature excitatory synaptic currents in developing rat neocortex
Journal of neurophysiology, 1993
1. In vitro slices of frontal neocortex were prepared from rat pups at various ages after birth: ... more 1. In vitro slices of frontal neocortex were prepared from rat pups at various ages after birth: postnatal days (PN) 3-5, 6-8, and 9-14. Using whole-cell patch-clamp techniques, both spontaneous and evoked excitatory postsynaptic currents (EPSCs) were recorded from voltage-clamped layer II-III pyramidal neurons. Developmental changes in EPSCs were examined. 2. Four properties of miniature EPSCs (mEPSCs) were studied: rise time, amplitude, decay time constant (tau), and frequency. These parameters were not tetrodotoxin sensitive and did not exhibit significant developmental changes during the first two postnatal weeks. 3. mEPSCs occurred approximately every 2-3 s and had peak amplitudes of 25-30 pA. Within each age group, certain parameters of mEPSCs were voltage dependent. mEPSC rise time and decay tau were significantly increased at depolarized potentials (-30 to -45 mV) relative to hyperpolarized (-75 to -90 mV) or resting membrane potential (RMP) (-60 to -70 mV). 4. At threshold ...
Picrotoxin-(PTX) induced epileptiform activity was studied in guinea pig hippocampal slices maint... more Picrotoxin-(PTX) induced epileptiform activity was studied in guinea pig hippocampal slices maintained in vitro, using intra- and extracellular recording techniques. The observed pattern of spontaneous and evoked epileptiform activity was quite complex. Spontaneous epileptiform events originated in the CA3 region and subsequently spread or propagated to CA1. Activation of CA1 could then reactivate CA3. This reverberation of activity was seen also following stimulation of the mossy fiber afferents from the dentate gyrus to CA3. Stimulation of fibers in the stratum radiatum of the CA1 region could trigger, at short latency, epileptiform activity that either was localized in CA1 or also occurred in CA3, with a late secondary discharge in CA1. This is attributed to a backfiring of the Schaffer collaterals and illustrates the ability of a variety of CA3 inputs to trigger epileptiform activity. Bath-applied PTX, at concentrations of 50-200 microM, had no apparent effect on the resting mem...
1. gamma-Aminobutyric acid (GABA) (10-500 microM) was applied to cultured chick cerebral neurons ... more 1. gamma-Aminobutyric acid (GABA) (10-500 microM) was applied to cultured chick cerebral neurons by pressure ejection, and the resulting currents (IGABA) were recorded using standard whole-cell voltage-clamp techniques. Plots of the peak IGABA as a function of membrane potential were nonlinear with an outwardly rectifying appearance. 2. IGABA decayed during a prolonged application of GABA. This decay was associated with a decline in the conductance of the cell, suggesting that the decline in IGABA was principally due to receptor desensitization. 3. After 5-7 days in culture, whole-cell recordings revealed the presence of spontaneous synaptic currents. These currents were presumed to be GABA-gated inhibitory postsynaptic currents (IPSCs) because they reversed at the Cl- equilibrium potential (ECl-), were blocked by picrotoxin (25 microM), and were prolonged by pentobarbital (50 microM). 4. Synaptic currents were analyzed by fitting exponential functions to their decay. In normal reco...
Conductance changes underlying a late synaptic hyperpolarization in hippocampal CA3 neurons
Journal of neurophysiology, 1987
1. Single-electrode current- and voltage-clamp techniques were employed to study properties of th... more 1. Single-electrode current- and voltage-clamp techniques were employed to study properties of the conductance underlying an orthodromically evoked late synaptic hyperpolarization or late inhibitory postsynaptic potential (IPSP) in CA3 pyramidal neurons in the rat hippocampal slice preparation. 2. Late IPSPs could occur without preceding excitatory postsynaptic potentials at the resting membrane potential and were graded according to the strength of the orthodromic stimulus. The membrane hyperpolarization associated with the late IPSP peaked within 140-200 ms after orthodromic stimulation of mossy fiber afferents. The late IPSP returned to base line with a half-decay time of approximately 200 ms. 3. As determined from constant-amplitude hyperpolarizing-current pulses, the membrane conductance increase during the late IPSP, and the time course of its decay, were similar whether measurements were made near the resting membrane potential or when the cell was hyperpolarized by approxima...
Intracellular injection of EGTA abolished the hyperpolarization seen after spontaneous and evoked... more Intracellular injection of EGTA abolished the hyperpolarization seen after spontaneous and evoked bursts of action potentials in hippocampal CA3 neurons. Simultaneously recorded, orthodromically elicited IPSPs were relatively unaffected by this treatment. It is concluded that CA3 neurons generate at least two types of hyperpolarization: one resulting from a calcium-mediated potassium conductance and the other from increased conductance to chloride.
Quisqualate induces an inward current via mGluR activation in neocortical pyramidal neurons
Brain Research, 2000
Activation of metabotropic glutamate receptors (mGluRs) has multiple effects on the excitability ... more Activation of metabotropic glutamate receptors (mGluRs) has multiple effects on the excitability of pyramidal neurons in rat frontal neocortex. Synaptic transmission and intrinsic excitability are both affected. During studies of the effects of quisqualate on synaptic activity, it was observed that quisqualate also induced a slow inward current. Whole-cell patch clamp recordings were obtained from layer II/III pyramidal neurons of neocortical slices in vitro. The bath solution contained APV, CNQX and bicuculline to block ionotropic glutamate and GABA(A) receptors. At a holding potential of -70 mV, quisqualate (2 microM) induced an inward current of about 60 pA. The response was reversible upon washing. This current was associated with an increase in membrane conductance and was still seen in the presence of TTX (0.5 microM). Bath application of the nonselective mGluR antagonist, (R, S)-alpha-methyl-4-carboxyphenyglycine (MCPG, 200-500 microM) reduced the current by 70%. Other mGluR agonists (ACPD, DHPG, L-CCG-1 and L-AP4) did not induce a significant inward current at the concentrations tested. The current-voltage relation of the quisqualate-induced current was linear with a reversal potential near 0 mV suggesting involvement of nonselective cation channels. The quisqualate-induced inward current was markedly reduced (72%) with 200 microM GDP-beta-S in the pipette solution, indicating that it is a postsynaptic phenomenon mediated by a G-protein dependent mechanism. These results suggest that mGluRs can directly increase the postsynaptic excitability of pyramidal cells.
Cultures of cerebral neurons were prepared from chick embryos, 8.5 days in ovo, and maintained in... more Cultures of cerebral neurons were prepared from chick embryos, 8.5 days in ovo, and maintained in vitro. Following chronic exposure of these cells to GABA, the levels of [ 3 H]flunitrazepam binding in situ and electrophysiological responsiveness to γ-aminobutyric acid (GABA) was ...
Zinc enhances GABAergic transmission in rat neocortical neurons
Journal of Neurophysiology
1. Intracellular recordings were made in layer II-III neurons of rat neocortical slices maintaine... more 1. Intracellular recordings were made in layer II-III neurons of rat neocortical slices maintained in vitro. The effect of bath application of zinc (50-300 microM) on evoked synaptic activity and passive membrane properties was examined. 2. Excitatory postsynaptic potentials (EPSPs) mediated by N-methyl-D-aspartate (NMDA) and non-NMDA receptors were recorded in response to electrical stimulation. Zinc did not affect either type of EPSP. Resting membrane potential, repetitive firing properties, and input resistance were not altered by zinc. 3. Inhibitory postsynaptic potentials (IPSPs) were enhanced after zinc application. Zinc also induced generation of large amplitude spontaneous gamma-aminobutyric acid-A (GABAA)- and GABAB-mediated IPSPs. Postsynaptic responses to iontophoretically applied GABA were unaffected. In the presence of zinc, GABAergic synaptic potentials could result in generation of action potentials. 4. Directly evoked IPSPs recorded in the presence of the excitatory ...
Layer I neurons of the rat neocortex. II. Voltage-dependent outward currents
Journal of Neurophysiology
1. Whole cell patch-clamp techniques, combined with direct visualization of neurons, were used to... more 1. Whole cell patch-clamp techniques, combined with direct visualization of neurons, were used to study voltage-dependent potassium currents in layer 1 neurons and layer II/III pyramidal cells. 2. In the presence of tetrodotoxin, step depolarizations evoked an outward current. This current had a complex waveform and appeared to be a composite of early and late components. The early peak of the composite K+ outward current was larger in layer I neurons. 3. In both layer I and pyramidal cells, the composite outward K+ current could be separated into two components based on kinetic and pharmacological properties. The early component was termed I(A) because it was a transient outward current activating rapidly and then decaying. I(A) was more sensitive to blocking by 4-aminopyridine (4-AP) than tetraethylammonium (TEA). The second component, termed the delayed rectifier or I(DR), activated relatively slowly and did not decay significantly during a 200-ms test pulse. I(DR) was insensitiv...
The Journal of neuroscience : the official journal of the Society for Neuroscience, Jan 18, 2015
Circuit dysfunction in complex brain disorders such as schizophrenia and autism is caused by imba... more Circuit dysfunction in complex brain disorders such as schizophrenia and autism is caused by imbalances between inhibitory and excitatory synaptic transmission (I/E). Short-term plasticity differentially alters responses from excitatory and inhibitory synapses, causing the I/E ratio to change as a function of frequency. However, little is known about I/E ratio dynamics in complex brain disorders. Transcriptional dysregulation in interneurons, particularly parvalbumin interneurons, is a consistent pathophysiological feature of schizophrenia. Peroxisome proliferator activated receptor γ coactivator 1α (PGC-1α) is a transcriptional coactivator that in hippocampus is highly concentrated in inhibitory interneurons and regulates parvalbumin transcription. Here, we used PGC-1α(-/-) mice to investigate effects of interneuron transcriptional dysregulation on the dynamics of the I/E ratio at the synaptic and circuit level in hippocampus. We find that loss of PGC-1α increases the I/E ratio ont...
Enhanced receptiveness at all synapses on a neuron that receive glutamatergic input is called cel... more Enhanced receptiveness at all synapses on a neuron that receive glutamatergic input is called cell-wide synaptic upscaling. We hypothesize that this type of synaptic plasticity may be critical for long-term memory storage within cortical circuits, a process that may also depend on epigenetic mechanisms, such as covalent chemical modification of DNA. We found that DNA cytosine demethylation mediates multiplicative synaptic upscaling of glutamatergic synaptic strength in cultured cortical neurons. Inhibiting neuronal activity with tetrodotoxin (TTX) decreased the cytosine methylation of and increased the expression of genes encoding glutamate receptors and trafficking proteins, in turn increasing the amplitude but not frequency of miniature excitatory postsynaptic currents (mEPSCs), indicating synaptic upscaling rather than increased spontaneous activity. Inhibiting DNA methyltransferase (DNMT) activity, either by using the small-molecule inhibitor RG108 or by knocking down Dnmt1 and ...
The cerebral cortex receives an extensive serotonergic (5-hydroxytryptamine, 5-HT) input. Immunoh... more The cerebral cortex receives an extensive serotonergic (5-hydroxytryptamine, 5-HT) input. Immunohistochemical studies suggest that inhibitory neurons are the main target of 5-HT innervation. In vivo extracellular recordings have shown that 5-HT generally inhibited cortical pyramidal neurons, whereas in vitro studies have shown an excitatory action. To determine the cellular mechanisms underlying the diverse actions of 5-HT in the cortex, we examined its effects on cortical inhibitory interneurons and pyramidal neurons. We found that 5-HT, through activation of 5-HT(2A) receptors, induced a massive enhancement of spontaneous inhibitory postsynaptic currents (sIPSCs) in pyramidal neurons, lasting for approximately 6 min. In interneurons, this 5-HT-induced enhancement of sIPSCs was much weaker. Activation of 5-HT(2A) receptors also increased spontaneous excitatory postsynaptic currents (sEPSCs) in pyramidal neurons. This response desensitized less and at a slower rate. In contrast, 5-H...
Early postnatal freeze lesions in rat neocortex produce anatomic abnormalities resembling those o... more Early postnatal freeze lesions in rat neocortex produce anatomic abnormalities resembling those observed in human patients with seizure disorders. Although in vitro brain slices containing the lesion are hyperexcitable, the mechanisms of this alteration have yet to be elucidated. To test the hypothesis that changes in postsynaptic inhibitory receptors may underlie this hyperexcitability, we examined properties of gamma-aminobutyric acid type A receptor (GABAAR)-mediated miniature inhibitory postsynaptic currents (mIPSCs). Recordings were obtained in layer II/III pyramidal cells located 1-2 mm lateral to the lesion. mIPSC peak amplitude and rate of rise were increased relative to nonlesioned animals, whereas decay time constant and interevent interval were unaltered. Bath application of zolpidem at a concentration (20 nM) specific for activation of the type 1 benzodiazepine receptor had no significant effect on decay time constant in six of nine cells. Exposure to higher concentratio...
The effect of metabotropic glutamate receptor (mGluR) activation on inhibitory synaptic transmiss... more The effect of metabotropic glutamate receptor (mGluR) activation on inhibitory synaptic transmission was examined by using whole cell patch-clamp recordings. Spontaneous (s) and miniature (m) inhibitory postsynaptic currents (IPSCs) were recorded from visually identified layer II/III pyramidal neurons in rat neocortex in vitro. Excitatory postsynaptic currents (EPSCs) were blocked by using bath application of 20 microM D(-)2-amino-5-phosphonovaleric acid and 10 microM 6-cyano-7-nitroquinoxaline-2,3-dione. In the presence of 1S,3R-1-aminocyclopentane-1,3-dicarboxylic acid (30-100 microM), Lp4-quisqualate (5 microM), and the group I selective mGluR agonist (S)-3,5-dihydroxyphenylglycine (100 microM), the frequency of sIPSCs was increased. Decay kinetics of sIPSCs were unaffected. No enhancement of mIPSCs was observed. Bath application of group II (2S,3S,4S-alpha-carboxycyclopropyl-glycine; 5 microM) and group III selective mGluR agonists (L-2-amino-4-phosphonobutyric acid; 100 microM)...
With the use of the whole cell patch-clamp technique combined with visualization of neurons in br... more With the use of the whole cell patch-clamp technique combined with visualization of neurons in brain slices, we studied the properties of miniature excitatory postsynaptic currents (mEPSCs) in rat neocortical layer I neurons. At holding potentials (-50 to -70 mV) near the resting membrane potential (RMP), mEPSCs had amplitudes of 5-100 pA and were mediated mostly by alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionate (AMPA) receptors. Amplitude histograms were skewed toward large events. An N-methyl-D-aspartate (NMDA) component was revealed by depolarization to -30 mV or by the use of a Mg2+-free bathing solution. At RMP, averaged AMPA mEPSCs had a 10-90% rise time of approximately 0.3 ms (uncorrected for instrument filtering). The decay of averaged mEPSCs was best fit by double-exponential functions in most cases. The fast, dominating component had a decay time constant of approximately 1.2 ms and comprised approximately 80% of the total amplitude. A small slow component had a de...
1. Whole-cell patch clamp techniques were used to record transient outward potassium currents in ... more 1. Whole-cell patch clamp techniques were used to record transient outward potassium currents in embryonic rat neocortical neurons maintained in culture. The effect of lidocaine and its quaternary derivative QX222 on this transient outward current (TOC) was examined. 2. Extracellular application of lidocaine produced a progressive decrease in peak TOC amplitude with no change in the overall current waveform. Peak amplitude was reduced to 74 +/- 6% (mean +/- SE) of control by 0.5 mM and to 48 +/- 2% by 2 mM lidocaine. The effect of lidocaine was fully reversible. Intracellular application of QX222 also resulted in a concentration-dependent reduction of the TOC with 0.5 mM reducing the peak amplitude to 73 +/- 8% of control and 1 mM reducing it to 41 +/- 4%. 3. Lidocaine reduced TOCs when applied intracellularly but the rate of block was considerably slower than with extracellular application. QX222 had no effect on the TOC when applied extracellularly. 4. Lidocaine (1 mM) induced a 4...
Developmental changes in NMDA and non-NMDA receptor-mediated synaptic potentials in rat neocortex
Journal of neurophysiology, 1993
1. In vitro slices of frontal neocortex were prepared from rat pups 3-14 days of age. Whole-cell ... more 1. In vitro slices of frontal neocortex were prepared from rat pups 3-14 days of age. Whole-cell patch-clamp recordings were obtained from layer II-III cortical neurons, and measurements of passive membrane properties were made. The development of evoked synaptic excitation and inhibition was also examined with the use of current- and voltage-clamp techniques. 2. Pharmacological separation of excitatory synaptic activity into both N-methyl-D-aspartate (NMDA) and non-NMDA receptor-mediated components was accomplished by application of D(-)2-amino-5-phosphonovaleric acid (APV), D(-)2-amino-7-phosphonoheptanoic acid (AP7), and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). Inhibitory synaptic events were described according to their reversal potentials and modulation by the GABAA receptor antagonist bicuculline methiodide (BMI). 3. Pups were grouped into three categories on the basis of age: postnatal day (PN) 3-5, PN 6-8, and PN 9-14. In slices from PN 3-5 pups, neurons exhibited high i...
NMDA receptor-mediated components of miniature excitatory synaptic currents in developing rat neocortex
Journal of neurophysiology, 1993
1. In vitro slices of frontal neocortex were prepared from rat pups at various ages after birth: ... more 1. In vitro slices of frontal neocortex were prepared from rat pups at various ages after birth: postnatal days (PN) 3-5, 6-8, and 9-14. Using whole-cell patch-clamp techniques, both spontaneous and evoked excitatory postsynaptic currents (EPSCs) were recorded from voltage-clamped layer II-III pyramidal neurons. Developmental changes in EPSCs were examined. 2. Four properties of miniature EPSCs (mEPSCs) were studied: rise time, amplitude, decay time constant (tau), and frequency. These parameters were not tetrodotoxin sensitive and did not exhibit significant developmental changes during the first two postnatal weeks. 3. mEPSCs occurred approximately every 2-3 s and had peak amplitudes of 25-30 pA. Within each age group, certain parameters of mEPSCs were voltage dependent. mEPSC rise time and decay tau were significantly increased at depolarized potentials (-30 to -45 mV) relative to hyperpolarized (-75 to -90 mV) or resting membrane potential (RMP) (-60 to -70 mV). 4. At threshold ...
Picrotoxin-(PTX) induced epileptiform activity was studied in guinea pig hippocampal slices maint... more Picrotoxin-(PTX) induced epileptiform activity was studied in guinea pig hippocampal slices maintained in vitro, using intra- and extracellular recording techniques. The observed pattern of spontaneous and evoked epileptiform activity was quite complex. Spontaneous epileptiform events originated in the CA3 region and subsequently spread or propagated to CA1. Activation of CA1 could then reactivate CA3. This reverberation of activity was seen also following stimulation of the mossy fiber afferents from the dentate gyrus to CA3. Stimulation of fibers in the stratum radiatum of the CA1 region could trigger, at short latency, epileptiform activity that either was localized in CA1 or also occurred in CA3, with a late secondary discharge in CA1. This is attributed to a backfiring of the Schaffer collaterals and illustrates the ability of a variety of CA3 inputs to trigger epileptiform activity. Bath-applied PTX, at concentrations of 50-200 microM, had no apparent effect on the resting mem...
1. gamma-Aminobutyric acid (GABA) (10-500 microM) was applied to cultured chick cerebral neurons ... more 1. gamma-Aminobutyric acid (GABA) (10-500 microM) was applied to cultured chick cerebral neurons by pressure ejection, and the resulting currents (IGABA) were recorded using standard whole-cell voltage-clamp techniques. Plots of the peak IGABA as a function of membrane potential were nonlinear with an outwardly rectifying appearance. 2. IGABA decayed during a prolonged application of GABA. This decay was associated with a decline in the conductance of the cell, suggesting that the decline in IGABA was principally due to receptor desensitization. 3. After 5-7 days in culture, whole-cell recordings revealed the presence of spontaneous synaptic currents. These currents were presumed to be GABA-gated inhibitory postsynaptic currents (IPSCs) because they reversed at the Cl- equilibrium potential (ECl-), were blocked by picrotoxin (25 microM), and were prolonged by pentobarbital (50 microM). 4. Synaptic currents were analyzed by fitting exponential functions to their decay. In normal reco...
Conductance changes underlying a late synaptic hyperpolarization in hippocampal CA3 neurons
Journal of neurophysiology, 1987
1. Single-electrode current- and voltage-clamp techniques were employed to study properties of th... more 1. Single-electrode current- and voltage-clamp techniques were employed to study properties of the conductance underlying an orthodromically evoked late synaptic hyperpolarization or late inhibitory postsynaptic potential (IPSP) in CA3 pyramidal neurons in the rat hippocampal slice preparation. 2. Late IPSPs could occur without preceding excitatory postsynaptic potentials at the resting membrane potential and were graded according to the strength of the orthodromic stimulus. The membrane hyperpolarization associated with the late IPSP peaked within 140-200 ms after orthodromic stimulation of mossy fiber afferents. The late IPSP returned to base line with a half-decay time of approximately 200 ms. 3. As determined from constant-amplitude hyperpolarizing-current pulses, the membrane conductance increase during the late IPSP, and the time course of its decay, were similar whether measurements were made near the resting membrane potential or when the cell was hyperpolarized by approxima...
Uploads
Papers by J. Hablitz