Biomedical subjects

F E Dudek

Publications and source records attributed to F E Dudek.

At least 73 records · Page 4Linked to original sources

PubMed1994

Blockade of NMDA-activated channels by magnesium in the immature rat hippocampus.

1. N-methyl-D-aspartate (NMDA)-receptor single-channel currents were examined in outside-out patches from thin slices of rat hippocampus to examine possible age dependence in the Mg(2+)-block of NMDA-mediated responses. The kinetics of Mg2+ block, among other channel properties, were compared between CA1 pyramidal cells from neonatal and 2-wk-old animals and dentate gyrus granule cells from neonatal and 4-wk-old animals. 2. Two distinct conductance states were seen consistently in nominally Mg(2+)-free solutions. The main conductance state was 50 and 56 pS in pyramidal cells and granule cells, respectively. The difference in NMDA-receptor channel conductance between the two cell types was statistically significant at all ages examined. Both cell types exhibited a less frequent 35- to 40-pS conductance state. 3. Channel closing rates showed no intrinsic voltage dependence in Mg(2+)-free solutions in any group. Open times became strongly voltage dependent when Mg2+ was added. The rate of block by Mg2+ was similar in all groups. 4. Voltage and Mg2+ influenced primarily the shortest time constant of shut-time distributions. Longer components varied relatively little with voltage or [Mg2+]. The effects of voltage and [Mg2+] suggest that brief shuttings represent closed states in the absence of Mg2+ and primarily a Mg(2+)-blocked state in the presence of Mg2+. 5. The rate of unblocking by Mg2+ was similar in all groups. Thus the dissociation constant for Mg2+ binding (i.e., the ratio of the unblocking and blocking rates) also showed little variation. 6. NMDA-receptor channels in two regions of the hippocampus behaved in a qualitatively similar fashion both in neonates and 2-or 4-wk-old rats. These observations are inconsistent with previous studies obtained using other methods, which suggested that block of NMDA receptor channels by Mg2+ increases substantially with age. Nevertheless, subtle developmental and regional differences in other aspects of NMDA-receptor channel behavior were detected, perhaps reflecting variations in molecular structure tailored to specific functional requirements.

Age Factors↗

PubMed1994

Age-related epileptogenic effects of corticotropin-releasing hormone in the isolated CA1 region of rat hippocampal slices.

1. The effects of corticotropin-releasing hormone (CRH) on synaptically evoked population and intracellular responses in the isolated rat CA1 region of hippocampal slices were studied to evaluate possible differences between adult and juvenile rats. 2. The amplitude of orthodromically evoked (stratum radiatum stimulation) population spikes was reversibly enhanced by 0.2-0.6 microM CRH to a greater extent in slices from juvenile rats than from adult rats. In no case, however, did CRH cause seizure-like activity to develop under normal recording conditions. 3. In the presence of 10-30 microM bicuculline, interictal-like bursts of population spikes and corresponding intracellularly recorded action potentials could be evoked starting at postnatal day 8. The number of spikes and the duration of the evoked bursts in the CA1 region were reversibly increased by CRH (0.2-0.6 microM) to a greater extent in slices from juvenile than from adult rats. 4. The amplitude of the afterhyperpolarization following intracellularly evoked bursts of action potentials in CA1 pyramidal cells was reduced by 0.2 microM CRH to a similar degree in both young and adult rats. No consistent changes in input resistance or membrane potential were observed. 5. No correlation was found between the magnitude of the CRH-induced increase in responsiveness and the initial excitability in controls, suggesting that the CRH-induced changes were independent of any age-dependent differences in general slice excitability. 6. Our results indicate that, in the CA1 region, CRH augments bicuculline-induced bursts to a greater extent in slices from young versus adult rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

PubMed1993

Neuronal synchronization without calcium-dependent synaptic transmission in the hypothalamus.

A critical question in understanding the mammalian brain is how populations of neurons become synchronized. This is particularly important for the neurons and neuroendocrine cells of the hypothalamus, which are activated synchronously to control endocrine glands and the autonomic nervous system. It is widely accepted that communication between neurons of the adult mammalian brain is mediated primarily by Ca(2+)-dependent synaptic transmission. Here we report that synchronous neuronal activity can occur in the hypothalamic suprachiasmatic nucleus without active Ca(2+)-dependent synaptic transmission. Simultaneous extracellular recordings of neuronal activity in the suprachiasmatic nucleus, which contains the mammalian biological clock, confirmed a circadian rhythm of synchronized activity in hypothalamic slices. Ca(2+)-free medium, which blocks chemical synaptic transmission and increases membrane excitability, produced periodic and synchronized bursts of action potentials in a large population of suprachiasmatic nucleus neurons with diverse firing patterns. N-Methyl-D-aspartic acid, non-N-methyl-D-aspartic acid, and gamma-aminobutyric acid type A receptor antagonists had no effect on burst synchrony. Whole-cell patch-clamp recordings confirmed that the Ca(2+)-free solution blocked evoked postsynaptic potentials and that the mixture of antagonists blocked the remaining spontaneous postsynaptic potentials. Therefore, mechanisms other than Ca(2+)-dependent synaptic transmission can synchronize neurons in the mammalian hypothalamus and may be important wherever neuronal networks are synchronized.

2-Amino-5-phosphonovalerate↗

PubMed1993

Cellular communication in the circadian clock, the suprachiasmatic nucleus.

The hypothalamic suprachiasmatic nucleus functions as the circadian clock in the mammalian brain. Communication between the cells of the suprachiasmatic nucleus is likely to be responsible for the generation and accuracy of this biological clock. Communication between many cells of the brain is mediated by action potentials that pass down the axon and cause release of neurotransmitters at the neuronal synaptic junction. Additional mechanisms of cellular communication appear to operate in the suprachiasmatic nucleus. Several lines of evidence point to multiple modes of cellular communication: these include the continuing operation of the clock after Na(+)-mediated action potentials have been blocked, the orchestrated metabolic rhythms of suprachiasmatic nucleus cells prior to synaptogenesis, the entrainment of fetal to maternal rhythms, and the rapid recovery of function after suprachiasmatic nucleus transplants into arrhythmic rodents. Possible alternative means of intercellular communication in the suprachiasmatic nucleus are examined, including calcium spikes in presynaptic dendrites, ephaptic interaction, paracrine communication, glial mediation, and gap junctions. This paper identifies and examines some of the unanswered questions related to intercellular communication of suprachiasmatic nucleus cells.

Action Potentials↗

PubMed1993

Membrane properties of identified guinea-pig paraventricular neurons and their response to an opioid mu-receptor agonist: evidence for an increase in K+ conductance.

Intracellular recordings were obtained from neurons in the paraventricular nucleus (PVN) of guinea-pig hypothalamic slices. Passive and active properties of the neurons were determined, and when possible, recorded neurons were injected with biocytin. The effects of a mu-receptor opioid agonist [D-Ala2, Nme-Phe4, Gly5-ol]enkephalin (DAGO) were studied in order to determine which types of cells have mu receptors and to test the hypothesis that an increase in K+ conductance causes mu-receptor-mediated inhibition in the PVN. The recorded cells inside the PVN were divided into two groups, primarily on the basis of the presence or absence of a low threshold Ca2+ spike (LTS). In one group of neurons, LTS potentials could not be evoked (non-LTS cells, n = 42). In another group of neurons (n = 35), LTS potentials with one or two Na+ spikes could be initiated with depolarizing pulses superimposed on steady hyperpolarizing currents; however, these neurons did not fire robust bursts (i.e. non-bursting LTS cells). The mean time constant of non-LTS cells (19.9 +/- 1.6 ms; mean +/- SEM) was significantly shorter than that of non-bursting LTS cells (26.7 +/- 2.1 ms). There were no differences in the mean resting membrane potential, spike amplitude, spike duration, input resistance, spike threshold and pattern of synaptic inputs between the two groups. Intracellular labeling with biocytin combined with cresyl violet counter-staining demonstrated that the two types of cells were located in the PVN.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Membrane properties of rat suprachiasmatic nucleus neurons receiving optic nerve input.

1. The electrophysiological properties of suprachiasmatic nucleus (SCN) neurons (n = 33) receiving optic nerve input were studied with intracellular recordings in rat hypothalamic slices maintained in vitro. Our major goal was to provide baseline data concerning the intrinsic membrane properties of these neurons and to test the hypothesis that the neurons are homogeneous electrophysiologically. 2. Action potentials were short in duration and followed by a pronounced hyperpolarizing after-potential. Spike amplitude (58.2 +/- 1.1 mV, mean +/- S.E.M.; measured from threshold), spike duration (0.83 +/- 0.03 ms; measured at half amplitude) and hyperpolarizing after-potential amplitude (23.9 +/- 1.0 mV; measured from threshold) appeared unimodally distributed and did not co-vary. 3. Intracellular injection of depolarizing current pulses evoked spike trains, and spike inactivation, spike broadening and frequency accommodation were always present. An after-hyperpolarization followed the spike train in all but one neuron. 4. Membrane time constant ranged from 7.5 to 21.1 ms (11.4 +/- 0.7 ms, n = 27), and its distribution appeared to be unimodal with the peak at approximately 10 ms. Input resistance ranged from 105 to 626 M omega (301 +/- 23 M omega, n = 33); the distribution also appeared unimodal with its peak at approximately 250 M omega. 5. A subpopulation (16 of 33, 48%) of the neurons exhibited slight (6-29%) time-dependent inward rectification in their voltage responses to hyperpolarizing current injection. Of the neurons lacking the time-dependent rectification, some (n = 5) exhibited time-independent inward rectification of 6-20% and others showed no (or < 3%) such rectification. The degree of inward rectification was correlated with neuronal excitability (r = 0.60, P < 0.002; assessed by measuring the steepness of the primary slope of the frequency-current plot) and with the spontaneous firing rate (r = 0.49, P < 0.007). Furthermore, the neurons with > 6% inward rectification (neurons with time-dependent and -independent rectification were combined) were more excitable (362 +/- 43 Hz/nA (n = 15) vs. 221 +/- 37 Hz/nA (n = 9), P < 0.05) and had a higher spontaneous firing rate (11.1 +/- 1.9 Hz (n = 19) vs. 3.9 +/- 1.5 Hz (n = 11), P < 0.02) than the neurons with no or negligible (i.e. < 3%) inward rectification. The two groups, however, were not significantly different in membrane time constant and input resistance. 6. When adequately hyperpolarized, twelve of seventeen (71%) neurons generated small low-threshold spike (LTS) potentials in response to depolarizing current pulses.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

PubMed1993

Local inhibitory synaptic inputs to neurones of the paraventricular nucleus in slices of rat hypothalamus.

1. Intracellular recordings were obtained from neurones in the region of the paraventricular nucleus in slices of rat hypothalamus. Glutamate microdrops were applied to the surface of the slices at sites dorsal, lateral and ventral to the paraventricular nucleus to selectively activate local presynaptic neurones. The gamma-aminobutyric acidA (GABAA)-receptor antagonists picrotoxin or bicuculline were bath-applied to block synaptic inhibition. 2. Glutamate microapplication caused a tonic depolarization and often repetitive action potentials in twenty of forty-seven recorded cells. This was probably caused by the direct exposure of the dendrites of the recorded cells to the glutamate microdrops. 3. Glutamate microstimulation elicited inhibitory synaptic responses in nine of forty-seven neurones tested. Glutamate microdrops caused discrete, hyperpolarizing postsynaptic potentials (PSPs) in four cells recorded with microelectrodes containing potassium acetate and evoked depolarizing PSPs in four cells recorded with KCl-filled microelectrodes. Glutamate microapplication inhibited spontaneous spike firing in another cell recorded with a potassium acetate microelectrode. 4. Bath application of GABAA-receptor antagonists completely blocked the hyperpolarizing PSPs elicited by glutamate microstimulation in three of three cells recorded with potassium acetate electrodes and the depolarizing PSPs in two of two cells recorded with KCl electrodes, indicating they were inhibitory PSPs caused by the release of GABA. Suppression of GABAA-mediated synaptic inhibition did not reveal any glutamate-evoked excitatory PSPs. 5. Recorded cells were identified as magnocellular, parvocellular or non-paraventricular bursting neurones on the basis of their electrophysiological properties. Direct depolarization and local inhibitory synaptic responses were observed in all three cell types. 6. Several conclusions can be drawn from these data: (1) functional glutamate receptors are distributed throughout neuronal populations in the paraventricular region of the hypothalamus, confirming and extending previous observations; (2) local synaptic inputs to neurones in the paraventricular nucleus are primarily inhibitory, supplied by perinuclear GABAergic neurones; (3) both magnocellular and parvocellular subpopulations receive local inhibitory synaptic inputs. The possibility that these local GABAergic circuits mediate inhibitory inputs to paraventricular neurones from limbic structures is discussed.

Animals↗

PubMed1993

Increased propensity for nonsynaptic epileptiform activity in immature rat hippocampus and dentate gyrus.

1. Low-[Ca2+] bursting was studied in hippocampal slices from immature and adult rats to test the hypothesis that the increased seizure susceptibility of the immature brain involves nonsynaptic mechanisms. Extracellular recordings were obtained from area CA1 of the hippocampus and from the dentate gyrus in slices from rats 6-9 days old (1 wk), 11-15 days old (2 wk), 19-23 days old (3 wk), and > 60 days old (adult). These slices were exposed to a low-[Ca2+] solution that included the calcium chelator, ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), and the excitatory amino acid antagonists, 6,7-dinitroquinoxaline-2,3-dione (DNQX) and DL-2-amino-5-phosphonopentanoic acid (AP-5). They were also exposed to a hyposmolar low-[Ca2+] solution (diluted with 20% H2O by volume), which induced or intensified the bursting. The propensity for nonsynaptic bursting and the characteristics of the bursts were compared between age groups. 2. The 1-wk group showed no bursting activity under any treatment condition in either CA1 or the dentate gyrus. Bursting occurred more frequently in the 2- and 3-wk groups than in the adult group in both CA1 and the dentate gyrus. 3. In CA1 the duration of the bursts was longer in the 2- and 3-wk groups as compared with the adult group. The number of population spikes per burst was also higher in slices from immature rats in dilute low-[Ca2+] solution. These findings demonstrate that nonsynaptic bursting in area CA1 is more robust in tissue from immature rats than adults.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Properties and synaptic mechanisms of bicuculline-induced epileptiform bursts in neocortical slices from children with intractable epilepsy.

1. Bicuculline-induced epileptiform bursts in slices of neocortical tissue resected from children (3 mo to 14 yr) undergoing neurosurgical treatment for intractable epilepsy were studied with conventional intracellular recording techniques. The purposes of this study were to characterize the bursts evoked in immature human neocortical slices, to gain further insight to how N-methyl-D-aspartate (NMDA) and non-NMDA receptors contribute to the genesis of the bursts, and to determine whether the characteristics of the bursts were related to patient age or clinically defined abnormality of the tissue. 2. Epileptiform bursts evoked by focal stimulation of the underlying white or gray matter in bicuculline (10 microM) were all-or-none events. Once evoked, the bursts in a given neuron appeared very similar to one another, regardless of stimulus intensity. Stronger stimuli only decreased the onset latency of the bursts. The bursts evoked with relatively weak stimuli (< 2-3 times the threshold), particularly those from stimulation of a distant site (4-5 mm), were variable in onset latency. The bursts from stimulation of a close site (0.5-2 mm) with stronger stimuli (> 3 times the threshold) were invariable in onset latency. 3. Across different cells, particularly across the cells in different slices, the bursts were quite variable in terms of their morphology and duration. When measured at one-half of the amplitude of the underlying depolarization (approximately 20-50 mV), the duration of the bursts ranged from 20 to 775 ms (n = 80 cells). In 23% of the cases (18 of 80 cells), afterdischarges lasting for tens of milliseconds to a few seconds followed the bursts.(ABSTRACT TRUNCATED AT 250 WORDS)

2-Amino-5-phosphonovalerate↗

PubMed1993

Patch-clamp analysis of spontaneous synaptic currents in supraoptic neuroendocrine cells of the rat hypothalamus.

Spontaneous synaptic currents were recorded in supraoptic magnocellular neurosecretory cells using whole-cell patch-clamp techniques in the rat hypothalamic slice preparation. Numerous spontaneous excitatory and inhibitory postsynaptic currents (EPSCs and IPSCs) were observed in the 27 cells recorded. The rate of occurrence of the spontaneous currents and the relative proportion of EPSCs versus IPSCs varied significantly from cell to cell. Miniature EPSCs and IPSCs were clearly distinguished from background noise in TTX (n = 3 cells at 0.5 micrograms/ml). The frequency of EPSCs and IPSCs decreased by approximately 70% and the largest events were blocked in TTX, but the peaks of the amplitude histograms were shifted by only a few picoamperes. Bicuculline (n = 10 cells at 10 microM and 2 cells at 20 microM) blocked completely all the IPSCs without any detectable effect on the frequency or amplitude of the EPSCs. No slow spontaneous outward currents, indicative of a K+ current from activation of GABAB receptors, were observed. The alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA)/kainate-type glutamate receptor antagonist 6-cyano-2,3-dihydroxy-7-nitroquinoxaline (CNQX; n = 7 cells at 10 microM) consistently blocked all EPSCs without any apparent effect on the frequency or amplitude of the IPSCs. No synaptic events could be detected when CNQX was applied in combination with bicuculline (n = 4). The decay phase of averaged spontaneous IPSCs and EPSCs recorded at resting membrane potential could be well fitted by single exponential functions in most cells. The time constants ranged from 0.92 to 3.0 msec for EPSCs (five cells) and from 5.3 to 6.6 msec for IPSCs (four cells). A second, slower time constant of 4-15 msec was found in the largest averaged EPSCs (> or = 40 pA). The amplitude of this slow component was -2 to -4 pA. These results suggest that, in the in vitro slice preparation, glutamate mediates all the spontaneous EPSCs in magnocellular neurosecretory cells by acting primarily on AMPA/kainate-type receptors at resting membrane potential and that activation of GABAA receptors mediates most if not all IPSCs.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed1993

Electrophysiology of the suprachiasmatic nucleus: synaptic transmission, membrane properties, and neuronal synchronization.

Knowledge of the neuronal membrane properties and synaptic physiology of the suprachiasmatic nucleus (SCN) is critical for an understanding of the cellular basis of circadian rhythms in mammals. The hypothalamic slice preparations from rodents and a combination of electrophysiological techniques (i.e., extracellular single- and multiple-unit recording, intracellular recording, and whole-cell patch clamp) were used to study (1) the role of excitatory and inhibitory amino acids (i.e., glutamate and gamma-aminobutyric acid [GABA] in synaptic transmission, (2) the membrane properties of SCN neurons, and (3) the mechanisms of neuronal synchronization. Antagonists for N-methyl-D-aspartate (NMDA) receptors and non-NMDA receptors blocked excitatory postsynaptic potentials (EPSPs) evoked by stimulation of the optic nerve or other sites when SCN cells were depolarized or at rest, respectively. Bicuculline blocked inhibitory postsynaptic potentials (IPSPs) that were evoked by local stimulation or that occurred spontaneously. The IPSP reversal potential was near the Cl- equilibrium potential, and was shifted to depolarized levels by raising intracellular [Cl-]. Thus, glutamate and GABA appear to mediate fast excitatory and inhibitory synaptic transmission in the SCN. Some SCN neurons, but not all of them, had low-threshold Ca2+ spikes and time-dependent inward rectification, thus indicating that the electrical properties of SCN neurons are not homogenous. Neurons with a firing rate of > 6 Hz had a regular pattern, and neurons with a rate of < 4 Hz had an irregular pattern; since both the firing rate and pattern could be modified with injected currents, SCN neurons with different firing patterns are unlikely to represent distinct classes of cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

Electrophysiology of dentate granule cells after kainate-induced synaptic reorganization of the mossy fibers.

Morphological data from humans with temporal lobe epilepsy and from animal models of epilepsy suggest that seizure-induced damage to dentate hilar neurons causes granule cells to sprout new axon collaterals that innervate other granule cells. This aberrant projection has been suggested to be an anatomical substrate for epileptogenesis. This hypothesis was tested in the present study with intra- and extracellular recordings from granule cells in hippocampal slices removed from rats 1-4 months after kainate treatment. In this animal model, hippocampal cell loss leads to sprouting of mossy fiber axons from the granule cells into the inner molecular layer of the dentate gyrus. Unexpectedly, when slices with mossy fiber sprouting were examined in normal medium, extracellular stimulation of the hilus or perforant path evoked relatively normal responses. However, in the presence of the GABAA-receptor antagonist, bicuculline, low-intensity hilar stimulation evoked delayed bursts of action potentials in about one-quarter of the slices. In one-third of the bicuculline-treated slices with mossy fiber sprouting, spontaneous bursts of synchronous spikes were superimposed on slow negative field potentials. Slices from normal rats or kainate-treated rats without mossy fiber sprouting never showed delayed bursts to weak hilar stimulation or spontaneous bursts in bicuculline. These data suggest that new local excitatory circuits may be suppressed normally, and then emerge functionally when synaptic inhibition is blocked. Therefore, after repeated seizures and excitotoxic damage in the hippocampus, synaptic reorganization of the mossy fibers is consistently associated with normal responses; however, in some preparations, the mossy fibers may form functional recurrent excitatory connections, but synaptic inhibition appears to mask these potentially epileptogenic alterations.

Action Potentials↗

PubMed1992

Osmolality and nonsynaptic epileptiform bursts in rat CA1 and dentate gyrus.

In several clinical situations, such as hyposmolar states and hypoxia-ischemia, reductions in the size of the extracellular space are associated with increased seizure susceptibility. Nonsynaptic interactions provide a likely means of mediating the effect of extracellular space on seizure susceptibility. Synchronous bursting of CA1 hippocampal neurons occurs via nonsynaptic mechanisms in solutions containing very low [Ca2+] and excitatory amino acid antagonists. We tested the hypothesis that lowering the osmolality of the extracellular medium could induce nonsynaptic bursting in the dentate gyrus, even though it is normally resistant to this treatment. Extracellular field potentials were recorded in the dentate gyrus and CA1 area of rat hippocampal slices. In the low-[Ca2+] solution with normal osmolality, bursts of population spikes were recorded from the dentate gyrus in only 7% of the slices, but solutions with decreased osmolality induced bursting in 63%. Corresponding values for the CA1 area were 60 and 73%, respectively. Mannitol, which reversed the hyposmolar state, abolished bursting in both regions. This study demonstrates that reducing the size of the extracellular space by lowering extracellular osmolality can transform a seizure-resistant area into one that exhibits robust epileptiform activity.

Action Potentials↗

PubMed1992

Intracellular electrophysiological study of suprachiasmatic nucleus neurons in rodents: inhibitory synaptic mechanisms.

1. The mechanisms responsible for evoked and spontaneous fast inhibitory postsynaptic potentials (IPSPs) in the hypothalamic suprachiasmatic nucleus (SCN) were studied with intracellular recording. SCN neurons, primarily ones identified as receiving excitatory optic nerve input, were recorded in rat and guinea-pig brain slice preparations maintained in vitro. 2. In normal medium, electrical stimulation of a site dorsocaudal to the SCN evoked IPSPs from thirty-three of thirty-six neurons. The evoked IPSPs rose to the peak quickly (8.7 +/- 0.9 ms, mean +/- S.E.M.; n = 15 neurons) and decayed gradually with a time constant of 25 +/- 3 ms. Spontaneous IPSPs were present in each of the thirty-six neurons. These IPSPs had a rise-to-peak time of 7.2 +/- 1.0 ms (n = 6 neurons) and a decay time constant of 14 +/- 5 ms. 3. When recorded with potassium acetate-filled electrodes, the evoked and spontaneous IPSPs were hyperpolarizing at resting membrane potential (less negative than -70 mV) and had a reversal potential of around -75 mV. On the other hand, when recorded with potassium chloride-filled electrodes, the IPSPs were depolarizing at membrane potentials more negative than -50 mV and had an estimated reversal potential less negative than spike threshold. 4. Bath application of bicuculline (10-50 microM), a gamma-aminobutyric acidA (GABAA) receptor antagonist, resulted in a complete blockade of both the evoked (n = 16) and spontaneous (n = 13) IPSPs. The bicuculline effects were reversible, and were not associated with any significant and consistent change in baseline membrane potential or input resistance. The neurons impaled in bicuculline-containing medium (n = 11) exhibited neither spontaneous IPSPs nor evoked IPSPs. 5. In some neurons bicuculline-resistant hyperpolarizing potentials, which were similar to the fast IPSPs in time course, occurred spontaneously or were evoked by electrical stimulation of the optic nerve or the dorsocaudal site. A fast prepotential always preceded the hyperpolarizing potential, and hyperpolarizing currents blocked these events, indicating that they were not synaptic in origin. No slow IPSPs were detected. 6. The results suggest that fast IPSPs from non-retinal afferents exist in virtually all SCN neurons receiving excitatory retinal input, and that GABAA receptors associated with Cl- channels mediate the fast IPSPs.

Action Potentials↗

PubMed1992

Single-electrode voltage-clamp analysis of the N-methyl-D-aspartate component of synaptic responses in neocortical slices from children with intractable epilepsy.

1. Synaptic transmission mediated by the N-methyl-D-aspartate (NMDA)-receptor type was studied in neocortex from children undergoing surgical treatment for intractable epilepsy. Intracellular recordings from pyramidal cells were obtained in slices of neocortical tissue by use of microelectrodes. Synaptic responses were induced by electrical stimulation and studied with current-clamp and single-electrode voltage-clamp techniques. The NMDA-receptor-mediated component of the synaptic responses was isolated by addition of 10 microM bicuculline and 30 microM 6-cyano-2,3-dihydroxy-7-nitroquinoxaline (CNQX) in the perfusion solution. 2. In the presence of bicuculline and CNQX, electrical stimulation evoked an excitatory postsynaptic potential (EPSP) in every recorded cell. The amplitude of this EPSP increased when membrane potential was depolarized with injected current. 3. All cells studied in voltage clamp were recorded with microelectrodes containing Cs+ and QX 314. To avoid contamination of the responses from voltage-dependent Ca2+ conductances, membrane potential was held at depolarized potentials until Ca2+ spiking inactivated completely. The evoked excitatory postsynaptic currents (EPSCs) measured at resting membrane potential ranged from 100 to 400 pA. The NMDA receptor-selective antagonist DL-2-amino-5-phosphonopentanoic acid (AP-5) reversibly decreased the current amplitude by 60% for 10 microM and 80% for 30 microM. 4. The current-voltage (I-V) relation showed a region of negative slope conductance between -100 and -20 mV. The largest currents (-250 to -900 pA) were recorded in the range of -45 to -20 mV and reversed between -10 and +10 mV. Removing Mg2+ from the perfusion solution decreased the negativity of the slope, which is consistent with a reduction in the voltage-dependent Mg2+ block of the NMDA-receptor channel. 5. The I-V plots obtained from cells recorded in the most abnormal tissue were averaged and compared with those from the least abnormal tissue. No significant difference was found between these two groups. The averaged plots from the youngest patients (8 and 10 mo old) and those from the oldest (5-15 yr old) patients were also compared, and the results from these two groups were not significantly different.(ABSTRACT TRUNCATED AT 400 WORDS)

2-Amino-5-phosphonovalerate↗

PubMed1992

Local synaptic circuits and epileptiform activity in slices of neocortex from children with intractable epilepsy.

1. Single and dual intracellular recordings were performed in neocortical slices obtained from tissue samples surgically removed from children (8 mo to 15 yr) for the treatment of intractable epilepsy. Electrical stimulation and glutamate microapplication were used to study local synaptic inputs to pyramidal cells. 2. In recordings with potassium-acetate electrodes, activation of presynaptic neocortical neurons with glutamate microdrops did not elicit a clear increase in postsynaptic potentials (PSPs) but did suppress current-evoked repetitive spike firing in recorded neurons. Bicuculline (10 microM) blocked this effect, suggesting it was caused by the activation of presynaptic gamma-aminobutyric acid (GABA) cells. In recordings with KCl electrodes, glutamate microdrops elicited an increase in the frequency and amplitude of depolarizing PSPs. Bicuculline (5-10 microM) blocked the glutamate-evoked PSPs, suggesting they were reversed GABAA-receptor-mediated inhibitory postsynaptic potentials (IPSPs). In one cell recorded with a KCl electrode (total n = 8), current-evoked spike trains elicited afterdischarges of reversed IPSPs, thus revealing a recurrent inhibitory circuit. Therefore local inhibitory synaptic circuits were robust and could be observed in tissue from patients as young as 11 mo. 3. In addition to short-latency (10-25 ms), monosynaptic excitatory postsynaptic potentials (EPSPs), electrical stimulation at low intensities sometimes elicited delayed EPSPs (20-60 ms). When GABAA-receptor-mediated synaptic inhibition was partially reduced in bicuculline (5-10 microM), electrical stimulation evoked large EPSPs at long and variable latencies (100-300 ms). Glutamate microapplication caused an increase in the frequency and amplitude of EPSPs; preliminary results suggest that glutamate microdrops were less likely to evoke EPSPs in tissue from younger patients (8-12 mo) than in slices from patients greater than 4 yr. Evidence for local excitatory synaptic circuits was thus found when synaptic inhibition was partially reduced. 4. After further reduction of inhibition in bicuculline (5-50 microM), electrical stimulation elicited epileptiform bursts. In pairs of simultaneously recorded neurons, bursts were generated synchronously from long-latency EPSPs (100-300 ms) in slices from patients as young as 8 mo. Reflected EPSPs at very long and variable latencies (500-1,100 ms) and repetitive epileptiform bursts could be evoked synchronously in pairs of cells. Glutamate activation of local presynaptic neurons elicited robust epileptiform events in recorded cells. This was seen in slices from patients as young as 16 mo. 5. These data provide physiological evidence for the presence of local inhibitory and excitatory synaptic circuits in human neocortex at least as early as 11 and 8 mo, respectively.(ABSTRACT TRUNCATED AT 400 WORDS)

Adolescent↗

PubMed1992

Prolonged field bursts in the dentate gyrus: dependence on low calcium, high potassium, and nonsynaptic mechanisms.

1. The dentate gyrus has been proposed to be a gate for entry of neuronal activity into the hippocampus. This function would give it a critical role in the propagation of seizure activity in that region. The hallmark of epileptiform activity in the dentate itself, often referred to as "maximal dentate activation" (MDA), has not been reproduced previously in vitro. 2. With the use of rat hippocampal slices, bath [Ca2+] was decreased, and [K+] was increased concurrently to simulate conditions found during intense neuronal activity in vivo. Both evoked and spontaneous field bursts were observed in the dentate granule cell layer under these conditions. These bursts were similar to MDA, consisting of a prolonged negative shift in extracellular potential with large-amplitude population spikes. 3. In 0.5 mM bath [Ca2+], single stimuli applied to the perforant path could evoke prolonged field bursts in the dentate only when bath [K+] was > or = 9 mM. However, repetitive stimulation (10 Hz) of the perforant path could elicit similar dentate responses when bath [K+] was as low as 5 mM. 4. In 0.5 mM bath [Ca2+], interictal-type bursts appeared spontaneously in CA1 and CA3 when bath [K+] was > or = 5 mM but were lost when [K+] was > 9 mM. Spontaneous seizurelike activity in the dentate required a higher minimum bath [K+] (9 mM) and persisted at [K+] of 11 mM. 5. Stimulation-evoked field bursts in the dentate altered epileptiform activity in CA3. At bath [K+] insufficient to cause spontaneous CA3 bursts, CA3 was activated transiently when prolonged field bursts occurred in the dentate. At higher bath [K+] in which spontaneous CA3 bursts did occur, they were depressed during the dentate bursts. 6. Deletion of Ca2+ from the bath; the addition of 30 microM each of bicuculline methiodide, D,L-2-amino-5-phosphonopentanoate (AP-5), and 6,7-dinitroquinoxaline-2,3-dione (DNQX); or the combination of both manipulations did not block antidromically evoked or spontaneous prolonged field bursts in the dentate. Thus the mechanisms maintaining and propagating these events did not require fast amino acid-mediated synaptic transmission. 7. The concurrent alteration of [K+] and [Ca2+] required to produce prolonged field bursts in the dentate underscores the positive feedback relationship between neuronal excitation and extracellular ionic concentrations, whereas the ability of synaptic stimulation to trigger nonsynaptic seizurelike events such as these prolonged field bursts may be relevant to the transition from interictal to ictal activity in vivo.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acids↗

PubMed1992

Persistent hyperexcitability in isolated hippocampal CA1 of kainate-lesioned rats.

1. Subcutaneous kainate injection in rats evoked acute seizures and led to cell loss in the hilus and areas CA1 and CA3, which resembled the pattern of hippocampal sclerosis often associated with temporal lobe epilepsy in humans. 2. Simultaneous intra- and extracellular recordings were performed in the stratum pyramidale of area CA1 while stimulating in the stratum radiatum close to the recording electrodes. Responses from control slices consisted of a brief excitatory postsynaptic potential (EPSP) with only one action potential, corresponding to a single extracellular population spike, followed by a clear biphasic inhibitory postsynaptic potential (IPSP). In slices from kainate-treated animals, however, stimulation evoked a prolonged EPSP, which often triggered multiple action potentials corresponding to multiple extracellular population spikes. 3. In slices from kainate-treated animals, the mean amplitude but not the duration of the stimulation-evoked IPSP was reduced. The extent of the kainate-induced loss of inhibition in area CA1 was highly variable. 4. Low concentrations of bicuculline in control slices led to a moderate hyperexcitability, which consisted of multiple population spikes and mirrored the responses observed in slices from kainate-treated animals in normal ACSF. Prolonged application of 10-30 microM bicuculline for > or = 30 min led to a much higher level of hyperexcitability, which was similar in slices from controls and kainate-treated rats. These findings are consistent with the hypothesis that the hyperexcitability of CA1 pyramidal neurons following kainate treatment is mainly due to decreased GABAA-receptor-mediated inhibition and that the loss of inhibition is only partial.(ABSTRACT TRUNCATED AT 250 WORDS)

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