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I Mody

Publications and source records attributed to I Mody.

At least 37 records · Page 2Linked to original sources
PubMed1999

Properties of single NMDA receptor channels in human dentate gyrus granule cells.

1. Cell-attached single-channel recordings of NMDA channels were carried out in human dentate gyrus granule cells acutely dissociated from slices prepared from hippocampi surgically removed for the treatment of temporal lobe epilepsy (TLE). The channels were activated by L-aspartate (250-500 nM) in the presence of saturating glycine (8 microM). 2. The main conductance was 51 +/- 3 pS. In ten of thirty granule cells, clear subconductance states were observed with a mean conductance of 42 +/- 3 pS, representing 8 +/- 2 % of the total openings. 3. The mean open times varied from cell to cell, possibly owing to differences in the epileptogenicity of the tissue of origin. The mean open time was 2.70 +/- 0.95 ms (range, 1.24-4.78 ms). In 87 % of the cells, three exponential components were required to fit the apparent open time distributions. In the remaining neurons, as in control rat granule cells, two exponentials were sufficient. Shut time distributions were fitted by five exponential components. 4. The average numbers of openings in bursts (1.74 +/- 0.09) and clusters (3.06 +/- 0.26) were similar to values obtained in rodents. The mean burst (6.66 +/- 0.9 ms), cluster (20.1 +/- 3.3 ms) and supercluster lengths (116.7 +/- 17.5 ms) were longer than those in control rat granule cells, but approached the values previously reported for TLE (kindled) rats. 5. As in rat NMDA channels, adjacent open and shut intervals appeared to be inversely related to each other, but it was only the relative areas of the three open time constants that changed with adjacent shut time intervals. 6. The long openings of human TLE NMDA channels resembled those produced by calcineurin inhibitors in control rat granule cells. Yet the calcineurin inhibitor FK-506 (500 nM) did not prolong the openings of human channels, consistent with a decreased calcineurin activity in human TLE. 7. Many properties of the human NMDA channels resemble those recorded in rat hippocampal neurons. Both have similar slope conductances, five exponential shut time distributions, complex groupings of openings, and a comparable number of openings per grouping. Other properties of human TLE NMDA channels correspond to those observed in kindling; the openings are considerably long, requiring an additional exponential component to fit their distributions, and inhibition of calcineurin is without effect in prolonging the openings.

Animals↗
PubMed1999

Modulation of synaptic GABAA receptor function by PKA and PKC in adult hippocampal neurons.

Several protein kinases are known to phosphorylate Ser/Thr residues of certain GABAA receptor subunits. Yet, the effect of phosphorylation on GABAA receptor function in neurons remains controversial, and the functional consequences of phosphorylating synaptic GABAA receptors of adult CNS neurons are poorly understood. We used whole-cell patch-clamp recordings of GABAA receptor-mediated miniature IPSCs (mIPSCs) in CA1 pyramidal neurons and dentate gyrus granule cells (GCs) of adult rat hippocampal slices to determine the effects of cAMP-dependent protein kinase (PKA) and Ca2+/phospholipid-dependent protein kinase (PKC) activation on the function of synaptic GABAA receptors. The mIPSCs recorded in CA1 pyramidal cells and in GCs were differentially affected by PKA and PKC. In pyramidal cells, PKA reduced mIPSC amplitudes and enhanced the fraction of events decaying with a double exponential, whereas PKC was without effect. In contrast, in GCs PKA was ineffective, but PKC increased the peak amplitude of mIPSCs and also favored double exponential decays. Intracellular perfusion of the phosphatase inhibitor microcystin revealed that synaptic GABAA receptors of pyramidal cells, but not those of GCs, are continually phosphorylated by PKA and conversely, dephosphorylated, most likely by phosphatase 1 or 2A. This differential, brain region-specific phosphorylation of GABAA receptors may produce a wide dynamic range of inhibitory synaptic strength in these two regions of the hippocampal formation.

Animals↗
PubMed1999

Decreased sensitivity to Group III mGluR agonists in the lateral perforant path following kindling.

The ability of the selective Group III mGluR agonist L-serine-O-phosphate (L-SOP) to inhibit lateral perforant path (LPP) evoked responses in the dentate gyrus was tested in hippocampal slices from commissurally-kindled rats 1-2 days after the last seizure, implanted controls, and fully-kindled rats rested for 28 days without stimulated seizures (28 days post-seizure, 28 dps). L-SOP was more potent in controls than kindled or 28 dps animals, decreasing the fEPSP slope with IC50s of 2.4 microM, 18.7 microM and 10.5 microM, respectively. Paired pulse facilitation (PPF, 50 ms) was comparable in control and kindled rats, but was markedly reduced in 28 dps rats, indicating increased release probability. Inhibition of the field excitatory postsynaptic potentials (fEPSP) by L-SOP was correlated with enhanced PPF in all groups, affirming a presynaptic site of action. At moderate levels of L-SOP-induced inhibition (20-60%), PPF showed significantly greater enhancement in 28 dps than in the other two groups. These results are interpreted as showing a functional reduction of the presynaptic inhibitory Group III mGluR (probably mGluR8) response in the LPP after kindling. Furthermore, PPF changes indicate that the kindled state may be associated with a long-lasting increase in the probability of release from LPP terminals, which may be temporarily masked or counterbalanced by recent seizures.

Animals↗
PubMed1999

Glutamatergic synapses onto hippocampal interneurons: precision timing without lasting plasticity.

In the hippocampal formation GABAergic inhibitory interneurons have a major role in the synchronization of neuronal activity and are involved in the generation of large-scale network oscillations. Thus, interneurons function as a 'clock' that dictates when principal cells fire during suprathreshold excitatory drive. Interneurons receive strong excitatory innervation from glutamatergic neurons and it has been much debated whether these synapses show mechanisms of long-term plasticity similar to those found at principal-cell synapses. Recent findings support the lack of conventional forms of LTP and LTD in most interneurons, partly owing to the distinct anatomical and neurochemical features of interneuronal excitatory synapses. The uncommon properties of excitatory synapses on interneurons might be required for their functioning as accurate and reliable neuronal oscillators.

Animals↗
PubMed1999

Casein kinase-II regulates NMDA channel function in hippocampal neurons.

Several second-messenger-regulated protein kinases have been implicated in the regulation of N-methyl-D-aspartate (NMDA) channel function. Yet the role of calcium and cyclic-nucleotide-independent kinases, such as casein kinase II (CKII), has remained unexplored. Here we identify CKII as an endogenous Ser/Thr protein kinase that potently regulates NMDA channel function and mediates intracellular actions of spermine on the channel. The activity of NMDA channels in cell-attached and inside-out recordings was enhanced by CKII or spermine and was decreased by selective inhibition of CKII. In hippocampal slices, inhibitors of CKII reduced synaptic transmission mediated by NMDA but not AMPA receptors. The dependence of NMDA receptor channel activity on tonically active CKII thus permits changes in intracellular spermine levels or phosphatase activities to effectively control channel function.

Animals↗
PubMed1999

Synaptic plasticity in kindling.

Kindling is an experimental model of epilepsy resulting from progressive activity-dependent changes in neuronal structure and function. During kindling, pathologic changes occur at various levels of organization of the nervous system, ranging from altered gene-expression in individual neurons to the loss of specific neuronal populations and the rearrangement of synaptic connections. This chapter summarizes recent findings about the long-lasting (plastic) nature of the alterations at the level of single neurons with emphasis on the role of altered excitatory and inhibitory amino acid receptors. The modified synaptic ligand-gated ion channels (i.e., "epileptic receptors") may ultimately be responsible for the kindling epileptogenesis.

Animals↗
PubMed1998

Increased number of synaptic GABA(A) receptors underlies potentiation at hippocampal inhibitory synapses.

Changes in synaptic efficacy are essential for neuronal development, learning and memory formation and for pathological states of neuronal excitability, including temporal-lobe epilepsy. At synapses, where there is a high probability of opening of postsynaptic receptors, all of which are occupied by the released transmitter, the most effective means of augmenting postsynaptic responses is to increase the number of receptors. Here we combine quantal analysis of evoked inhibitory postsynaptic currents with quantitative immunogold localization of synaptic GABA(A) receptors in hippocampal granule cells in order to clarify the basis of inhibitory synaptic plasticity induced by an experimental model of temporal-lobe epilepsy (a process known as kindling). We find that the larger amplitude (66% increase) of elementary synaptic currents (quantal size) after kindling results directly from a 75% increase in the number of GABA(A) receptors at inhibitory synapses on somata and axon initial segments. Receptor density was up by 34-40% and the synaptic junctional area was expanded by 31%. Presynaptic boutons were enlarged, which may account for the 39% decrease in the average number of released transmitter packets (quantal content). Our findings establish the postsynaptic insertion of new GABA(A) receptors and the corresponding increase in postsynaptic responses augmenting the efficacy of mammalian inhibitory synapses.

Animals↗
PubMed1998

The dynamics of synchronized neurotransmitter release determined from compound spontaneous IPSCs in rat dentate granule neurones in vitro.

1. The properties of GABAA receptor-mediated spontaneous IPSCs generated in hippocampal dentate granule neurones were analysed using whole-cell voltage-clamp techniques in order to explore the functional consequences of the low number (6-12) and close proximity of synaptic contacts made by single GABAergic interneurones. 2. Spontaneous IPSCs (sIPSCs) occurred with a frequency of 14.0 +/- 9.1 Hz (n = 31) and revealed a multi-modal positively skewed amplitude distribution (39.0 +/- 19.8 pA, median values). 3. The variance of 10-90% rise times and decay kinetics between IPSCs decreased with increasing peak amplitude. Larger amplitude events had significantly faster rise times, consistent with their site of generation being proximal to the soma. The decay kinetics of sIPSCs did not significantly change with amplitude. 4. Large amplitude sIPSCs occurred singularly or in discrete bursts, repeated regularly at low frequency. The rising phase of such sIPSCs were multi-phasic, composed of clear step-like inflections that were not a product of noise. The variability between the rising phase of individual sIPSCs was quantified by calculating their standard deviation, which produced fast rising (0.22 +/- 0.05 ms time to peak, n = 16) functions with half-widths of 0.38 +/- 0.10 ms, which declined to plateaux. 5. Computer simulations demonstrated that IPSCs with properties similar to those recorded experimentally could be generated by the linear summation of groups of temporally dispersed component events. Standard deviation functions of the rising phase of simulated IPSCs accurately described distributions of the temporal dispersion of unitary components. 6. The GABA uptake inhibitor (R)-N[4,4-bis(3-methyl-2-thienyl)but-3-enl-yl] nipecotic acid (tiagabine) (10 microM, n = 12) significantly prolonged the decay of mIPSCs (6.5 +/- 0.8 to 8.7 +/- 1.0 ms, median values) and sIPSCs (6.2 +/- 0.4 to 7.3 +/- 1.2 ms, median values), but failed to alter the frequency of occurrence, 10-90% rise times or peak amplitude of events. The application of flurazepam (30 microM, n = 7; 50 microM, n = 4) prolonged the decay of sIPSCs regardless of their amplitude. 7. These data indicate that sIPSCs are formed by the summation of unitary components that occur asynchronously and that GABA released from multiple sites has independent post-synaptic actions.

Animals↗
PubMed1998

Calcium-dependent inactivation of high-threshold calcium currents in human dentate gyrus granule cells.

1. Dentate gyrus granule cells acutely dissociated from hippocampal slices obtained from chronic temporal lobe epilepsy (TLE) patients displayed a high-voltage activated (HVA) Ca2+ conductance with a pronounced Ca2+-dependent inactivation. 2. Inactivation time constants and peak HVA Ca2+ current (ICa) amplitudes did not differ between perforated patch and whole-cell recordings without added exogenous Ca2+ buffers, indicating that the Ca2+-dependent characteristics of ICa inactivation were well preserved in whole-cell recordings. 3. Inactivation time constants correlated with whole-cell ICa, and were increased when Ca2+ was replaced with Ba2+ in the external solution or 5 mM BAPTA was added to the pipette solution. 4. In recordings without added exogenous Ca2+ buffers, the time course of ICa inactivation was comparable between human TLE and kindled rat granule cells. Conversely, the time course of ICa in human TLE granule cells loaded with 5 mM intracellular BAPTA resembled that observed in buffer-free recordings from control rat neurones. 5. The loss of a putative intraneuronal Ca2+ buffer, the Ca2+-binding protein calbindin (CB), from human granule cells during TLE may result in the pronounced Ca2+-dependent ICa inactivation. This process could serve a neuroprotective role by significantly decreasing Ca2+ entry during prolonged trains of action potentials known to occur during seizures.

Adult↗
PubMed1998

The absence of a major Ca2+ signaling pathway in GABAergic neurons of the hippocampus.

The Ca2+/calmodulin-dependent protein phosphatase 2B or calcineurin (CN) participates in several Ca2+-dependent signal transduction cascades and, thus, contributes to the short and long term regulation of neuronal excitability. By using a specific antibody to CN, we demonstrate its absence from hippocampal interneurons and illustrate a physiological consequence of such CN deficiency. Consistent with the lack of CN in interneurons as detected by immunocytochemistry, the CN inhibitors FK-506 or okadaic acid significantly prolonged N-methyl-D-aspartate channel openings recorded in the cell-attached mode in hippocampal principal cells but not those recorded in interneurons. Interneurons were also devoid of Ca2+/calmodulin-dependent protein kinase IIalpha, yet many of their nuclei contained the cyclic AMP-responsive element binding protein. On the basis of the CN and Ca2+/calmodulin-dependent protein kinase IIalpha deficiency of interneurons, entirely different biochemical mechanisms are expected to govern Ca2+-dependent neuronal plasticity in interneurons versus principal cells.

Animals↗
PubMed1998

Ion channels in epilepsy.

There are specific alterations in the structure or function of ion channels in the epileptic brain. Some of these alterations may promote hyperexcitability, whereas others may protect neurons from the deleterious effects of epileptic discharges. With the use of human tissue resected from epilepsy patients and the comparison of cellular properties to those found in well-defined experimental models, we will continue to gain insight into the specific ion channel changes associated with epilepsies. Further genetic studies will help to elucidate the altered molecular mechanisms underlying ion channel changes in this devastating neurological disorder (Noebels, 1996). Whether it is a change in structure, function, or both, the study of ion channels in epilepsies will soon reveal specific characteristics of ion channels found only in epileptic tissue. If the altered properties of such ion channels cannot be found in control (nonepileptic) neurons, these channels might be called "epileptic" ion channels. An understanding of the specific structure, function, and pharmacology of these "epileptic" channels will yield important clues for future therapeutical approaches aimed at preventing epileptogenesis, and insight into the processes whereby ion channels become "epileptic" may finally open the way to prophylactic treatments of the epilepsies.

Epilepsy↗
PubMed1998

Calbindin-D28k fails to protect hippocampal neurons against ischemia in spite of its cytoplasmic calcium buffering properties: evidence from calbindin-D28k knockout mice.

Cytoplasmic calcium-binding proteins are thought to shield neurons against damage induced by excessive Ca2+ elevations. Yet, in theory, a mobile cellular Ca2+ buffer could just as well promote neuronal injury by facilitating the rapid dispersion of Ca2+ throughout the cytoplasm. In sharp contrast to controls, in mice lacking the gene for calbindin-D28k, synaptic responses of hippocampal CA1 pyramidal neurons which are normally extremely vulnerable to ischemia, recovered significantly faster and more completely after a transient oxygen-glucose deprivation in vitro, and sustained less cellular damage following a 12 min carotid artery occlusion in vivo. Other cellular and synaptic properties such as the altered adaptation of action potential firing, and altered paired-pulse and frequency potentiation at affected synapses in calbindin-D28k-deficient mice were consistent with a missing intraneuronal Ca2+ buffer. Our findings provide direct experimental evidence against a neuroprotective role for calbindin-D28k.

Animals↗
PubMed1998

Enhanced propagation of epileptiform activity through the kindled dentate gyrus.

Extracellular recordings were performed in combined hippocampal-entorhinal cortex (HC-EC) slices obtained from control and commissural kindled rats to investigate the propagation of epileptiform activity from the entorhinal cortex (EC) to the hippocampus (HC) after chronic epilepsy. Lowering extracellular Mg2+ concentration in control slices induced epileptiform activity consisting of spontaneous epileptiform bursts in area CA3 and of electrographic seizures in the EC. In contrast, the CA3 region of HC-EC slices obtained from kindled rats displayed significantly longer lasting epileptiform bursts and electrographic seizures. The electrographic seizures that were absent in controls propagated from the EC because disconnecting the HC from the EC stopped their occurrence in the CA3, whereas epileptiform bursts persisted with an unaltered pattern and frequency. Thus the area CA3 is affected by kindling and contributes to the spread of epileptiform activity within the EC-HC complex. We developed a method to induce focal epileptiform activity in the EC by locally perfusing the gamma-aminobutyric acid-A (GABA) antagonist bicuculline (50 mM) in 10 mM KCl containing artificial cerebrospinal fluid. This method enabled us to investigate the propagation of epileptiform discharges from the disinhibited EC to the DG without affecting the DG with the epileptogenic medium. We show here that kindling facilitates the propagation of epileptiform activity through the DG. These data are consistent with the normal function of the DG as a filter limiting the spread of epileptiform activity within the HC-EC complex. This gating mechanism breaks down after chronic epilepsy induced by kindling.

Animals↗
PubMed1998

Substance P enhances NMDA channel function in hippocampal dentate gyrus granule cells.

Substance P (SP)-containing afferents and the NK-1 tachykinin receptor to which SP binds are present in the dentate gyrus of the rat; however, direct actions of SP on principal cells have not been demonstrated in this brain region. We have examined the effect of SP on N-methyl--aspartate (NMDA) channels from acutely isolated dentate gyrus granule cells of adult rat hippocampus to assess the ability of SP to regulate glutamatergic input. SP produces a robust enhancement of single NMDA channel function that is mimicked by the NK-1-selective agonist Sar9, Met(O2)11-SP. The SP-induced prolongation of NMDA channel openings is prevented by the selective NK-1 receptor antagonist (+)-(2S, 3S)-3-(2-methoxybenzylamino)-2-phenylpiperidine (CP-99,994). Calcium influx or activation of protein kinase C were not required for the SP-induced increase in NMDA channel open durations. The dramatic enhancement of excitatory amino acid-mediated excitability by SP places this neuropeptide in a key position to gate activation of hippocampal network activity.

Animals↗
PubMed1997

Synaptic communication among hippocampal interneurons: properties of spontaneous IPSCs in morphologically identified cells.

The properties of spontaneous IPSCs (sIPSCs) recorded with whole-cell patch-clamp techniques were investigated in various anatomically identified hippocampal CA1 interneurons and were compared with those recorded in pyramidal cells. Neurons labeled with biocytin or neurobiotin were classified on the basis of their dendritic and axonal arborizations, leading to the identification of previously unknown interneuron types projecting to the dendritic region of pyramidal cells. In most interneurons, the average sIPSCs decayed slower than did those observed in pyramidal cells. The properties of sIPSCs were homogeneous within a given morphologically identified neuron type. Many interneurons had comparable somatic size, location, and dendritic arbor but displayed extremely different axonal projections paralleled by distinct sIPSC properties. Thus, physiological comparisons are only meaningful after the complete morphological identification of the recorded cells. The decay of sIPSCs matched for amplitudes and rise times could vary over 10-fold in a given interneuron, consistent with electrotonic filtering and possibly with different GABAA receptor subunit assemblies present at distinct synapses. Our findings demonstrate an extensive connectivity among hippocampal interneurons through GABAA synapses of various properties that may underlie complex network oscillations at different frequencies.

Action Potentials↗
PubMed1997

Silent GABAA synapses during flurazepam withdrawal are region-specific in the hippocampal formation.

Whole-cell patch-clamp recordings were made from CA1 pyramidal and dentate gyrus granule cells (GCs) in hippocampal slices to assess the effects of withdrawal from chronic flurazepam (FRZ) treatment on the function of synaptic GABAA receptors. In slices from control rats, acute perfusion of FRZ (30 microM) increased the monoexponential decay time constant of miniature IPSCs (mIPSCs) in CA1 and GCs (from 3.4 +/- 0.6 to 7.6 +/- 2.1 msec and from 4.2 +/- 0. 6 to 7.1 +/- 1.8 msec, respectively) but did not change their mean conductance, 10-90% rise time, or frequency of occurrence. Withdrawal (2-5 d) from chronic in vivo FRZ treatment (40-110 mg/kg per day, per os) resulted in a dramatic loss of mIPSCs in CA1 neurons. On day 5 of withdrawal, no mIPSCs could be recorded in 40% of CA1 pyramidal cells. In the remaining 60% of the neurons, mIPSCs had a reduced mean conductance (from 0.78 +/- 0.12 nS in vehicle-treated controls to 0.31 +/- 0.05 nS) and a diminished frequency of occurrence (from 20.7 +/- 7.9 to 4.1 +/- 0.6 Hz). We have estimated that >80% of GABAA synapses on CA1 pyramidal cells had become silent, whereas at still-active synapses the number of functional GABAA receptor channels decreased by 60%. This reduction rapidly reverted to control levels on day 6 of withdrawal. FRZ withdrawal did not alter mIPSC properties in GCs. Our results are consistent with the hypothesis that chronic benzodiazepine treatment leads to a reduced number of functional synaptic GABAA receptors in a region-specific manner that may stem from differences in the subunit composition of synaptic GABAA receptors.

Animals↗
PubMed1997

Endogenous GABA activates small-conductance K+ channels underlying slow IPSCs in rat hippocampal neurons.

The objective of this study was to determine the properties of K+ channels activated by endogenously released trasmitter under synaptic conditions. First, the levels of gamma-aminobutyric acid (GABA) were depleted in hippocampal nerve endings to establish the relative contribution of endogenously released GABA to the activation of GABA(B) receptors mediating slow inhibitory postsynaptic currents (IPSCs). Inhibition of glutamic acid decarboxylase and GABA reuptake effectively depleted >85% of the releasable GABA pool, producing parallel reductions of GABA(A) and GABA(B) receptor-mediated IPSCs, indicating that both classes of receptors are activated synaptically by endogenously released GABA. Whole cell patch-clamp recordings of stimulus-evoked slow IPSCs at potentials hyperpolarized from the potassium reversal potential were consistent with the activation of a nonrectifying (n = 3) or slightly outwardly rectifying (n = 4) K+ conductance by the endogenously released GABA. Spectral analysis of the decay phase of GABA(B) IPSCs revealed several time constants indicating complex underlying channel kinetics. Nonstationary variance analysis yielded a small unitary conductance in the range of 5-13 pS, consistent with a large number of channels activated during evoked currents. These results indicate that in granule cells of the dentate gyrus, GABA released synaptically from interneuron terminals activates an unusually small K+ conductance, with no or slight outward rectification. This conductance is therefore unlike those typically reported for neuronal G protein-coupled K+ channels or those activated by exogenously applied baclofen with larger, inwardly rectifying conductances.

Analysis of Variance↗