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K Krnjević

Publications and source records attributed to K Krnjević.

At least 19 recordsLinked to original sources
PubMed1995

Econazole, a blocker of Ca2+ influx, selectively suppresses LTP of EPSPs in hippocampal slices.

Econazole--an agent that suppresses Ca2+ influx triggered by depletion of internal Ca-stores in non-excitable cells--was bath-applied to submerged slices from Sprague-Dawley rats. Econazole (15-20 microM) had no consistent effect on afferent volleys, EPSPs or population spikes; but in seven out of nine slices, it prevented the induction of tetanic long-term potentiation (LTP) of field EPSPS. By contrast, all slices showed a marked LTP of population spikes. Ca2+ influx induced by tetanic depletion of Ca2+ stores may be essential for LTP of excitatory synaptic transmission.

Animals↗
PubMed1995

Internal Ca2+ stores involved in anoxic responses of rat hippocampal neurons.

1. During whole-cell recordings from CA1 neurons of rat brain slices with electrodes containing only KMeSO4 and Hepes, brief anoxia (2-3 min) consistently evoked a hyperpolarization (delta V approximately 14 mV) and reduction in input resistance (delta R approximately -20%). 2. As in previous intracellular recordings, Dantrolene sodium (10 microM) suppressed the anoxic delta V and delta R, confirming the release of internal Ca2+ is a major component of the anoxic response. 3. To identify the relevant intracellular Ca2+ store, other blockers of Ca2+ release were applied either externally (in the bath) or internally, by addition to the contents of the recording electrode. 4. The anoxic hyperpolarization was abolished or much reduced by heparin (10-20 micrograms ml-1, internal), thapsigargin (10 microM, external), Ruthenium Red (50 microM, internal) and external procaine (0.5-2 mM), but not by internal procaine (0.5-1 mM) or ryanodine (10 microM, external). 5. The anoxic fall in resistance was also abolished or reduced by heparin, thapsigargin and external procaine, but not by ryanodine, internal procaine or Ruthenium Red. 6. In addition, external procaine (0.5-2 mM) eliminated the early (transient) depolarization and reduced the post-anoxic hyperpolarization by 60 +/- 22%. 7. None of these agents consistently changed the resting potential, but the input resistance was significantly increased by Dantrolene and external procaine. 8. In view of the marked effects of heparin and thapsigargin, but not ryanodine and internal procaine, we conclude that the anoxic response seen in such whole-cell recordings is initiated predominantly by Ca2+ release from an internal store that is InsP3 sensitive rather than Ca2+ sensitive. 9. Comparable but less pronounced effects of external procaine were seen during intracellular recordings with 3 M KCl-containing electrodes. The dose-dependent suppression of various features of the anoxic response by external procaine (EC50 approximately 0.2 mM) is presumed to be mediated by a superficial membrane trigger or modulating site.

Animals↗
PubMed1995

Adenosine antagonists have differential effects on induction of long-term potentiation in hippocampal slices.

How adenosine leakage and tetanic release might affect long-term potentiation (LTP) was investigated by applying adenosine antagonists 8(p-sulfophenyl)theophylline (8SPT) or 8-cyclopentyl-3,7-dihydro-1,3-dipropyl-1H-purine-2,6-dione (DPCPX) to slices, while recording CA1 field EPSPs and population spikes. In the first series of experiments, we applied weak double tetani (at 100 Hz, for 1 s) that were subliminal for evoking LTP in initial control runs. In the presence of 8SPT--at concentrations (10-50 microM) which block both A1 and A2 receptors--the same tetani consistently evoked LTP of population spikes but not of excitatory postsynaptic potentials (EPSPs), whereas DPCPX (50 nM), which blocks only A1 receptors, facilitated LTP of both EPSPs and population spikes. These results are consistent with previous evidence that tetanic adenosine release on the one hand depresses LTP via A1 receptors but on the other facilitates LTP via A2 receptors. In a second set of experiments, 8SPT (50-100 microM) did not prevent the induction of LTP of both EPSPs and population spikes by stronger tetanic stimulation. Therefore A2 receptor activation is not essential for the induction of LTP when stronger tetani are applied. Overall, the main effect of endogenous adenosine release is to oppose LTP induction.

Adenosine↗
PubMed1995

Actions of diazoxide on CA1 neurons in hippocampal slices from rats.

Membrane effects of diazoxide (DZX) were examined in CA1 pyramidal neurons, mainly by whole-cell recording in slices kept at 33 degrees C (from Sprague-Dawley rats). Bath applications of DZX (0.65 mM) did not significantly change the resting input conductance; but instantaneous inward rectification was reduced by 47 +/- 14% (near -110 mV). There was a similar depression of a large, sustained voltage-dependent outward current (by 44 +/- 11% near 0 mV). A nearly identical reduction of the outward current recorded in a Ca current suppressing medium (but not in 30 mM tetraethylammonium) indicated that the DZX-sensitive current includes the delayed rectifier. In Mn, low-Ca medium containing tetraethylammonium and carbachol, DZX potentiated (by 43 +/- 12%) the D-type slowly decaying outward current seen after hyperpolarizing pulses at a holding potential of approximately -50 mV. DZX abolished or depressed slow inward currents, such as the tetrodotoxin-sensitive persistent Na current, high voltage activated Ca currents (IC50 = 0.47 mM), and the Q current. In 6 of 13 cells recorded with electrodes containing either guanosine or adenosine diphosphate, DZX potentiated the voltage-dependent outward current, but input conductances were reduced. In conclusion, although there was little indication that it activates classical KATP channels in CA1 neurons, DZX strongly depresses several voltage-dependent, slowly inactivating outward and inward currents, which are important modulators of cell excitability.

Animals↗
PubMed1994

Tolbutamide suppresses slow and medium afterhyperpolarization in hippocampal slices.

Afterhyperpolarizations (AHPs) were recorded (in whole-cell mode, with KMeSO4-containing electrodes) after multiple spikes evoked with 200 ms current pulses. Bath applications of tolbutamide (0.5-1 mM) to 12 CA1 neurones nearly abolished the medium and slow component of AHPs. Thus the AHPs generated by 7-8 spikes (mostly) were reduced by 82.6 +/- 5.2% (s.e.m.) at the initial peak (mAHP) and by 85.0 +/- 5.8% 1 s later (sAHP). Glibenclamide (10 microM) had no comparable blocking effect. The previous finding that tolbutamide (but not glibenclamide) selectively suppresses the anoxic hyperpolarization of CA1 neurones is therefore consistent with the idea that the anoxic hyperpolarization is mediated by Ca-dependent (especially AHP-type) K channels.

Animals↗
PubMed1994

Guanosine diphosphate is required for activation of a glyburide, ATP and cromakalim-sensitive outward current in rat hippocampal neurones.

Voltage-dependent outward currents were examined in CA1 neurones by whole-cell recording in slices. Nine cells were recorded with the 'standard' internal solution (KMeSO4, HEPES, EGTA, CaCl2, and MgCl2) and held a potential (-54 +/- 3 mV) at which there was no significant outward current. Cromakalim (CROM, 100 microM) reduced both input conductance (GN) (by 14 +/- 4%) and outward currents, evoked over a wide range of potentials by brief depolarizing pulses: at -4.0 +/- 3.0 mV, currents diminished by 30 +/- 10%. When 1 mM GDP was added to the standard internal solution, there was a significant outward current at approximately -54 mV; and CROM greatly increased outward currents near -4.0 mV (by 99 +/- 26.4%, n = 10). The enhanced outward currents were reduced by CROM washout (in two cells) and by 10 microM glyburide (GLYB, in four cells). When six other cells were recorded with electrodes containing both ATP (5 mM) and GDP (1 mM), there was no net outward current at approximately -54 mV and CROM reduced outward currents (at approximately 0 mV, by 37.5 +/- 10.9%). We conclude that GDP in hippocampal neurones appears to activate an ATP- and GLYB-sensitive outward current, which is much potentiated by CROM.

Action Potentials↗
PubMed1994

Anoxia selectively depresses excitatory synaptic transmission in hippocampal slices.

EPSPs/IPSPs were recorded with intracellular electrodes from CA1 neurons close to site of stimulation. Brief anoxia (3 min) abolished EPSPs but reduced IPSPs by 64.8 +/- 4.0% (n = 10); the remaining IPSP was presumed to be monosynaptic. The effects of anoxia on purely monosynaptic IPSPs were examined after pharmacological blockade of excitatory synaptic transmission with 2 mM kynurenate or 20 microM CNQX + 20 microM APV. In these tests, after 3 min of anoxia the slopes of IPSPs vs. membrane potential were reduced by only 38.2 +/- 4.3% (n = 12). The present study demonstrates that, contrary to previous reports, inhibitory synaptic transmission is quite resistant to anoxia.

2-Amino-5-phosphonovalerate↗
PubMed1994

Endogenous adenosine deaminase does not modulate synaptic transmission in rat hippocampal slices under normoxic or hypoxic conditions.

Field and intracellular potentials were recorded from CA1 pyramidal stratum in submerged slices (at 33 degrees). During "normal" oxygenation (95% O2 + 5% CO2), tonic depression of population spikes and field excitatory postsynaptic potentials by endogenous adenosine was demonstrated by (i) the marked enhancement by the adenosine antagonists 8-(p-sulfophenyl)theophylline (10 microM) and caffeine (0.2 mM), (ii) depression by the transport blocker dipyridamole (5 microM), and (iii) enhancement by exogenous adenosine deaminase (all tested by bath application). Thus, adenosine deaminase (0.5 units/ml) reduced by 10.7 +/- 3.0% (S.E.) the half-maximal stimulus intensity (for population spikes). The effects of adenosine deaminase were prevented by the specific inhibitor, deoxycoformycin (30 microM). In intracellular recordings, excitatory postsynaptic potentials were enhanced in a comparable manner by adenosine deaminase. By contrast, neither deoxycoformycin (5 and 30 microM) nor erythro-9-(2-hydroxy-3-nonyl)adenine (another adenosine deaminase inhibitor; 10 and 50 microM) had significant effects on population spikes. Superfusion with anoxic medium (saturated with 95% N2 + 5% CO2) for 2-3 min suppressed population spikes reversibly, by a mechanism involving adenosine, because 8-(p-sulfophenyl)theophylline (10 microM) and caffeine (0.2 mM) delayed the onset of anoxic block and accelerated the subsequent recovery, and the recovery was much slower or incomplete in the presence of dipyramidole (0.5 microM). However, the anoxic suppression of population spikes was not affected by deoxycoformycin (30 microM) or erythro-9-(2-hydroxy-3-nonyl)adenine (10 microM); the corresponding 50% postanoxic recovery times were also unchanged (e.g. 4.0 +/- 0.2 min for controls and 4.1 +/- 0.3 min in deoxycoformycin).(ABSTRACT TRUNCATED AT 250 WORDS)

Adenine↗
PubMed1994

Actions of cromakalim on outward currents of CA1 neurones in hippocampal slices.

1. Membrane effects of cromakalim (Crom; 50-300 microM) were examined in CA1 neurones recorded mainly by intracellular, single-electrode voltage-clamping in slices (from Sprague-Dawley rats) kept in an interface chamber at 33 degrees C. 2. In 14 cells held at -63 +/- 3.5 mV, in the presence of tetrodotoxin, kynurenic acid and (in most cases) bicuculline, bath applied Crom produced no consistent change in holding current (-59 +/- 66 pA) or input conductance (GN) (-3.9 +/- 5.2%). 3. Overall there were no significant changes in instantaneous inward rectification or in Q-current inward relaxations. 4. In 18 out of 22 cells, outward currents, evoked by 0.5 s pulses to voltages > -50 and < -20 mV, were depressed by Crom (by 42 +/- 11%, for n = 22). Because this effect was consistently seen in Ca current-blocking media, containing either Mn and low Ca, or Cd (and also carbachol), the K channels depressed by Crom were probably of the delayed rectifier (IDR) type. 5. The Crom-control difference current (ICrom), obtained with slow depolarizing ramps, had a biphasic character, inward in the voltage (V) range > -50 < -20 mV (where outward currents are depressed by Crom) and tending outward for V > or = -20 mV. 6. In 10 out of 11 cells, Crom potentiated a D-like, slowly-inactivating outward current (by 88 +/- 31%, for n = 11). 7 The effects of Crom and of 2 min periods of anoxia were compared in 12 cells: unlike anoxia, Cromproduced no consistent increases in GN; the currents evoked in the same cells by anoxia differed significantly from those evoked by Crom (by 150 +/- 60 pA); the directions of current changes induced byCrom and anoxia respectively were not significantly correlated. Crom strongly depressed anoxic outward currents (by 80 +/- 12%, n = 4).8 Some Crom-induced effects (increases in D-like current and the outward current elicited at V>- 20 mV) were always reversed by tolbutamide (1 mM), but much less consistently by glibenclamide(10-30 microM).9 In conclusion, the effects of Crom, recorded with intracellular electrodes in CA1 neurones in slices,show little resemblance to the effects of activation of ATP-sensitive K channels.

Animals↗
PubMed1993

Effects of trifluoperazine on synaptically evoked potentials and membrane properties of CA1 pyramidal neurons of rat hippocampus in situ and in vitro.

The effects of trifluoperazine (TFP), a phenothiazine antipsychotic, on hippocampal activity were studied in the CA1 subfield, both in situ and in slices. In the extracellular studies in situ and in vitro, both somatic population spikes and dendritic excitatory postsynaptic potentials (EPSP) fields were depressed reversibly by TFP, applied by microiontophoresis or in the bath (50-100 microM). Similar effects were also seen during iontophoretic applications of sphingosine in situ. Like TFP (at micromolar concentrations) sphingosine is a dual Ca2+/calmodulin-dependent kinase and protein kinase C (PKC) inhibitor. In intracellular recordings from slices, 50-100 microM TFP induced a slow depolarization and a decrease in input resistance (RN), probably through a gamma-aminobutyric acid (GABA)-mediated increase in Cl- conductance (GCl). TFP also reduced the slow afterhyperpolarization (AHP) as well as electrically evoked inhibitory postsynaptic potentials (IPSPs), but EPSPs were augmented in both amplitude and duration. When CA1 neurons were voltage clamped, TFP elicited a corresponding inward current (consistent with depolarization), increased the leak conductance, and enhanced excitatory synaptic currents; whereas inhibitory synaptic currents and high-threshold Ca2+ currents were reduced. In conclusion, these effects of TFP--which cannot be readily explained by its potent antidopamine action--are in keeping with other evidence that both Ca2+/calmodulin-dependent kinase and PKC can modulate GCl-conductance and high-threshold Ca(2+)-conductance, as well as inhibitory and excitatory postsynaptic currents.

Animals↗
PubMed1992

Cellular and synaptic actions of general anaesthetics.

1. This paper briefly reviews mechanisms by which such widely-used volatile anaesthetics as halothane and isoflurane suppress neural function in the brain. 2. In general, anaesthetics tend to depress neuronal firing and excitatory synaptic transmission, and potentiate synaptic inhibition. 3. According to recent evidence, a particular important action of anaesthetics is to inactivate a variety of both voltage-dependent and agonist-triggered Ca-currents. 4. Activation of K outward currents and Na inward currents probably occurs only with higher doses of anaesthetics. 5. How anaesthetics interfere with Ca-channels remains largely a matter of speculation--though some evidence favours a Ca-mediated action, following Ca2+ release from internal stores, that may account also for potentiation of IPSPs by prolonging the opening of GABA-activated Cl- channels. 6. Whatever its precise underlying mechanism, a suppression of Ca-influx into pre-synaptic terminals could well account for the depression of excitatory synaptic transmission.

Anesthesia, General↗
PubMed1992

Developmental and regional differences in the vulnerability of rat hippocampal slices to lack of glucose.

Field excitatory postsynaptic potentials were recorded in stratum radiatum of CA1 and CA3 in submerged hippocampal slices from adult or newborn (postnatal days 5-25) Wistar rats. In adult slices, excitatory postsynaptic potentials were depressed by glucose removal ("aglycemia") more rapidly and to a greater extent in CA1 than in CA3 [respective mean times to 50% reduction in peak amplitude were 7.5 +/- 0.83 (standard error) min and 12.5 +/- 0.27 (standard error) min]. Subsequent recovery of excitatory postsynaptic potentials in normoglycemic medium was correspondingly quicker in CA3 than in CA1. Transmission failure at the synapses was indicated by the preservation of the afferent volley, and sharp depression of synaptic input-output plots. In the early postnatal period, CA1 excitatory postsynaptic potentials were much more resistant to aglycemia, substantially persisting for as long as 75 min, with full subsequent recovery in normoglycemic medium. The higher resistance of slices from newborn rats progressively disappeared over the first two postnatal weeks.

Aging↗
PubMed1992

Glibenclamide depresses the slowly inactivating outward current (ID) in hippocampal neurons.

Sulphonylurea drugs, such as glibenclamide and tolbutamide, are widely used as selective blockers of adenosine triphosphate-sensitive K channels. In experiments on hippocampal slices (from Wistar rats) glibenclamide (and possibly gliquidone and tolbutamide) significantly reduced the highly voltage-dependent, 4-aminopyridine-sensitive D-type outward current of CA3 neurons. Judging by these observations, the sulphonylureas may not be as selective as generally believed.

Animals↗
PubMed1991

Anoxic block of GABAergic IPSPs.

In rat hippocampal slices GABAergic IPSPs are very rapidly suppressed by anoxia (in less than 2 min). Both early (GABAA) and late (GABAB) components are affected. After reoxygenation, the IPSPs recover, but only slowly and not always completely. Iontophoretic applications of GABA or baclofen indicated no major depression of responses during anoxia. It is therefore unlikely that the anoxic suppression of IPSPs is caused by desensitizations of GABA receptors. A more probable explanation is a failure of GABAergic neurons to release GABA from inhibitory nerve terminals.

Action Potentials↗
PubMed1991

Persistent pulsatile release of glutamate induced by N-methyl-D-aspartate in neonatal rat hippocampal neurones.

1. Intracellular recordings were made from CA3 hippocampal neurones in vitro, during the first ten days of postnatal life and in adulthood. 2. Repeated (three to six) applications of N-methyl-D-aspartate (NMDA), in the presence of tetrodotoxin (TTX, 1-3 microM) and K+ channel blockers (tetraethylammonium chloride or bromide (TEA), 10 mM, and Cs+, 2 mM; or 4-aminopyridine (4-AP), 30-50 microM, and Cs+, 2 mM) induced in neonatal but not in adult neurones, periodic inward currents (PICs) which persisted for several hours after the last application of NMDA. 3. PICs which were due to non-specific cation currents had a frequency of 0.10 +/- 0.04 Hz, and an amplitude of 1.1 +/- 0.28 nA at holding potentials between -40 and -50 mV. The amplitude was a linear function of the membrane potential over the range -70 to +20 mV. They reversed polarity at 4.1 +/- 9.8 mV. 4. K+ channel blockers alone failed to induce PICs. Repeated (three to six) brief applications of high (12 mM) K+ medium also induced PICs. The frequency and amplitude of K(+)-induced PICs were however considerably reduced by concomitant applications of the NMDA receptor antagonist D,L-3-[( +/- )-2-carboxypiperazin-4-yl-]propyl-1-phosphonic acid (CPP, 20 microM). PICs could be induced also by caffeine (1 mM) in the presence of the phosphodiesterase inhibitor 3-isobutyl-1-methyl-xanthine (IBMX, 200 microM), TTX, TEA and Cs+. 5. Intracellular injection of the calcium chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) did not prevent the induction of PICs by NMDA. However PICs were blocked by removal of the external calcium and by the calcium antagonists cobalt (2 mM) and cadmium (50 microM). 6. In spite of blockade of propagated synaptic activity by TTX, PICs were synchronous in a pair of intracellularly recorded cells. They were also synchronous with extracellular spikes recorded by electrodes located into stratum pyramidal or stratum radiatum. 7. Once established, PICs were unaffected by NMDA receptor antagonists D(-)2-amino-5-phosphonovaleric acid (AP-5, 50 microM), CPP (20 microM) and the NMDA channel blocker ketamine (10 microM). They were reversibly blocked by the broad spectrum excitatory amino acid antagonist kynurenic acid (1 mM) and by the selective non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM). 8. It is concluded that PICs are generated in neonatal neurones by a synchronous, pulsatile release of glutamate from presynaptic nerve terminals, secondary to oscillations in intracellular calcium.

Action Potentials↗
PubMed1991

Temperature dependence of extracellular ionic changes evoked by anoxia in hippocampal slices.

1. Extracellular [K] and [Ca] were measured with ion-selective microelectrodes in CA1 pyramidal cell layer of rat hippocampal slices in an interface chamber. 2. Near room temperature (21-22 degrees C), brief periods of anoxia (3- to 4-min substitution of 95% N2-5% CO2 for 95% O2-5% CO2) produced very small changes in [K]o [-0.022 +/- 0.10 (SE) mM] or [Ca]o (-0.030 +/- 0.0029 mM) and were associated with only minor depression of population spikes (-22.5 +/- 11%). 3. Stratum radiatum (SR) stimulation (0.2-5 Hz) could evoke substantial increases in [K]o (by 0.2-2 mM); although variable, they were consistent in any one slice. The same stimulation regularly caused only small depressions of [Ca]o (by less than 0.1 mM, typically). 4. Also at 21-22 degrees neither stimulation nor anoxia generated more than minimal reductions in extracellular space [by 2.3 +/- 0.94%, as measured by the tetramethylammonium (TMA) method], and spreading depression (SD) occurred in only 1 out of 20 slices. 5. At 33-34 degrees C, anoxia (also for 3-4 min) consistently produced more substantial increases in [K]o (0.83 +/- 0.18 mM); but the apparent changes in [Ca]o at 33 degrees C (0.058 +/- 0.12 mM) could not with certainty be distinguished from thermoelectric artifacts. There was a severe depression of population spikes (-76 +/- 10%). 6. Although electrical stimulation evoked greater reductions in [Ca]o, increases in [K]o were 50% smaller. 7. During anoxia at 33-34 degrees C, the extracellular space was significantly reduced, by 6.1 +/- 0.9%. Moreover, in 37% of the slices, either stimulation or anoxia triggered massive increases in [K]o (greater than 10 mM) and large reductions in [Ca]o (less than 1 mM), associated with SD-like swings in focal potential. 8. It is concluded that the extracellular ionic changes evoked by brief anoxia do not contribute in a major way to the depression of synaptic transmission.

Action Potentials↗
PubMed1990

Muscarinic actions in hippocampus are probably not mediated by cyclic GMP.

It has been proposed that, in a variety of tissues, guanosine 3':5'-monophosphate (cyclic GMP) is the intracellular mediator of muscarinic effects. This hypothesis was tested in the CA1 region of the hippocampus, in urethane-anaesthetized rats, by studying extracellularly muscarinic disinhibition of disfacilitation and the effect of dibutyryl cyclic GMP, muscarinic agents and an inhibitor of cyclic nucleotide-dependent kinase (H-8), all applied by microiontophoresis. The main findings were: (a) cyclic GMP analogues do not mimic disfacilitation or disinhibition produced by muscarinic agents; (b) N-(2-(methylamino)ethyl)-5-isoquinoline sulfonamide (H-8) does not prevent the excitatory actions of muscarinic agents; and (c) H-8 alone does not change the field responses. In conclusion, cyclic nucleotide-dependent kinases do not seem to play a major role in the on-going modulation of excitability in the hippocampus and cyclic GMP is unlikely to be a major intracellular messenger mediating directly or indirectly the excitatory actions of acetylcholine.

8-Bromo Cyclic Adenosine Monophosphate↗