Intracellular Injection Of Cs Research Articles

Role of potassium conductances in determining input resistance of developing brain stem motoneurons.

The role of potassium conductances in determining input resistance was studied in 166 genioglossal (GG) motoneurons using sharp electrode recording in brain stem slices of the rats aged 5-7 days, 13-15 days, and 19-24 days postnatal (P). A high magnesium (Mg(2+); 6 mM) perfusate was used to block calcium-mediated synaptic release while intracellular or extracellular cesium (Cs(+)) and/or extracellular tetraethylammonium (TEA) or barium (Ba(2+)) were used to block potassium conductances. In all cases, the addition of TEA to the high Mg(2+) perfusate generated a larger increase in both input resistance (R(n)) and the first membrane time constant (tau(0)) than did high Mg(2+) alone indicating a substantial nonsynaptic contribution to input resistance. With intracellular injection of Cs(+), GG motoneurons with lower resistance (<40 MOmega), on the average, showed a larger percent increase in R(n) than cells with higher resistance (>40 MOmega). There was also a significant increase in the effect of internal Cs(+) on R(n) and tau(0) with age. The largest percent increase (67%) in the tau(0) due to intracellular Cs(+) occurred at P13-15, a developmental stage characterized by a large reduction in specific membrane resistance. Addition of external Cs(+) blocked conductances (further increasing R(n) and tau(0)) beyond those blocked by the TEA perfusate. Substitution of external calcium with 2 mM barium chloride produced a significant increase in both R(n) and tau(0) at all ages studied. The addition of either intracellular Cs(+) or extracellular Ba(2+) created a depolarization shift of the membrane potential. The amount of injected current required to maintain the membrane potential was negatively correlated with the control R(n) of the cell at most ages. Thus low resistance cells had, on the average, more Cs(+)- and Ba(2+)-sensitive channels than their high resistance counterparts. There was also a disproportionately larger percent increase in tau(0) as compared with R(n) for both internal Cs(+) and external Ba(2+). Based on a model by Redman and colleagues, it might be suggested that the majority of these potassium conductances underlying membrane resistance are initially located in the distal dendrites but become more uniformly distributed over the motoneuron surface in the oldest animals.

Ionic mechanisms of muscarinic depolarization in entorhinal cortex layer II neurons.

The mechanisms underlying direct muscarinic depolarizing responses in the stellate cells (SCs) and non-SCs of medial entorhinal cortex layer II were investigated in tissue slices by intracellular recording and pressure-pulse applications of carbachol (CCh). Subthreshold CCh depolarizations were largely potentiated in amplitude and duration when paired with a short DC depolarization that triggered cell firing. During Na+ conductance block, CCh depolarizations were also potentiated by a brief DC depolarization that allowed Ca2+ influx and the potentiation was more robust in non-SCs than in SCs. Also, in non-SCs, CCh depolarizations could be accompanied by spikelike voltage oscillations at a slow frequency. In both SCs and non-SCs, the voltage-current (V-I) relations were similarly affected by CCh, which caused a shift to the left of the steady-state V-I relations over the entire voltage range and an increase in apparent slope input resistance at potentials positive to about -70 mV. CCh responses potentiated by Ca2+ influx demonstrated a selective increase in slope input resistance at potentials positive to about -75 mV in relation to the nonpotentiated responses. K+ conductance block with intracellular injection of Cs+ (3 M) and extracellular Ba2+ (1 mM) neither abolished CCh depolarizations nor resulted in any qualitatively distinct effect of CCh on the V-I relations. CCh depolarizations were also undiminished by block of the time-dependent inward rectifier Ih, with extracellular Cs . However, CCh depolarizations were abolished during Ca2+ conductance block with low-Ca2+ (0.5 mM) solutions containing Cd2+, Co2+, or Mn2+, as well as by intracellular Ca2+ chelation with bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid. Inhibition of the Na+-K+ ATPase with strophanthidin resulted in larger CCh depolarizations. On the other hand, when NaCl was replaced by N-methyl-D-glucamine, CCh depolarizations were largely diminished. CCh responses were blocked by 0.8 microM pirenzepine, whereas hexahydro-sila-difenidolhydrochloride,p-fluoroanalog (p-F-HHSiD) and himbacine were only effective antagonists at 5- to 10-fold larger concentrations. Our data are consistent with CCh depolarizations being mediated in both SCs and non-SCs by m1 receptor activation of a Ca2+-dependent cationic conductance largely permeable to Na+. Activation of this conductance is potentiated in a voltage-dependent manner by activity triggering Ca2+ influx. This property implements a Hebbian-like mechanism whereby muscarinic receptor activation may only be translated into substantial membrane depolarization if coupled to postsynaptic cell activity. Such a mechanism could be highly significant in light of the role of the entorhinal cortex in learning and memory as well as in pathologies such as temporal lobe epilepsy.

Electrogenic pump and a Ca(2+)- dependent K+ conductance contribute to a posttetanic hyperpolarization in lamprey sensory neurons.

1. Tetanic stimulation of lamprey sensory dorsal cells resulted in a posttetanic hyperpolarization (PTH). The amplitude and duration of the PTH were dependent on the stimulus duration and frequency. The PTH was not reversed at membrane potentials negative to -100 mV, whereas the afterhyperpolarization following single action potentials reversed at approximately -85 mV. There was also a biphasic effect on the input resistance during the PTH, with an early reduction that recovered to control before the PTH had decayed. 2. The amplitude and duration of the PTH were increased in Ringer solution containing tetraethylammonium and 4-aminopyridine, both of which broadened single action potentials, but were reduced after intracellular injection of Cs+. Ca(2+)-free Ringer solution, Cd2+, and Co2+ also reduced the PTH, suggesting the involvement of a Ca(2+)-dependent K+ conductance. However, the PTH was not reduced in Ba2+ Ringer solution, or by the Ca(2+)-dependent K+ channel antagonists apamin and charybdotoxin. 3. The cardiac glycoside ouabain reduced the amplitude and duration of the PTH, as did substitution of Na+ with choline or Li+. K(+)-free Ringer solution also reduced the PTH, whereas high-K+ Ringer solution had more variable effects. The amplitude and duration of the PTH were also dependent on temperature. These results support the involvement of an ouabain-sensitive Na-K pump in the PTH. 4. The PTH was reduced by the tachykinins substance P and physalaemin, and by 5-hydroxytryptamine, which blocks apamin-sensitive Ca(2+)-dependent K+ channels in the lamprey. However, gamma-aminobutyric acid, which has been reported to reduce a Ca(2+)-dependent K+ conductance in the dorsal cells, did not reduce the PTH. 5. These results suggest that a Ca(2+)-dependent K+ conductance and an Na-K electrogenic pump underlie the PTH. The PTH reduces the excitability of the dorsal cells, suggesting that it may act as a mechanism to gate sensory information entering the spinal cord.

Paired‐pulse depression of the N‐methyl‐D‐aspartate receptor‐mediated synaptic potentials in the amygdala

1. An in vitro slice preparation of rat amygdala was used to study the paired-pulse depression of the N-methyl-D-aspartate (NMDA) receptor-mediated synaptic potential e.p.s.p.NMDA. 2. The e.p.s.p.NMDA was isolated pharmacologically by applying a solution containing the non-NMDA receptor antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and the gamma-aminobutyric acidA (GABAA) blocker picrotoxin and increasing the stimulus intensity. 3. When two stimuli of identical strength were applied in close succession, the second e.p.s.p.NMDA was depressed. This paired-pulse depression was seen with interstimulus intervals of between 100 ms and 2000 ms; the maximal depression was observed at interval of 200 ms. 4. Superfusion of phaclofen or 2-hydroxy-saclofen inhibited the paired-pulse depression indicating the involvement of GABAB receptors. 5. Bath applications of Ba2+ or intracellular injection of Cs+ to block post- but not presynaptic GABAB receptors failed to inhibit the paired-pulse depression (PPD). 6. Incubation of slices with pertussis toxin prevented the postsynaptic hyperpolarization induced by baclofen. The PPD of e.p.s.p.NMDA, however, was not affected by pertussis toxin treatment. 7. These results suggest that GABA released by the first stimulus acts on GABAB receptors to suppress the second e.p.s.p.NMDA via mechanisms other than activation of a postsynaptic GABAB receptor-coupled K+ conductance.

Mechanisms of oscillatory activity in guinea‐pig nucleus reticularis thalami in vitro: a mammalian pacemaker.

1. The ionic mechanisms of rhythmic burst firing and single spike, tonic discharge were investigated with extracellular and intracellular recordings of single neurones in the guinea-pig nucleus reticularis thalami (NRT) maintained as a slice in vitro. 2. Activation of cortical/thalamic afferents to NRT neurones resulted in a short latency burst of action potentials which could be followed by a rhythmic sequence of oscillatory burst firing. Intracellularly, this oscillatory activity was associated with an alternating sequence of low threshold Ca2+ spikes separated by after-hyperpolarizing potentials. Intracellular injection of short duration hyperpolarizing current pulses resulted in a similar sequence of oscillatory burst firing, suggesting that this activity is an intrinsic property of NRT cells. The frequency of rhythmic burst firing was highly voltage and temperature dependent and was between 7-12 Hz at -65 to -60 mV at 38 degrees C. In addition, at depolarized membrane potentials, oscillatory burst firing was typically followed by a prolonged tail of single spike activity. 3. Application of the Na+ channel poison tetrodotoxin blocked the generation of fast action potentials, but left intact the rhythmic sequence of low threshold Ca2+ spikes separated by after-hyperpolarizing potentials (AHPs). The reversal potential of the AHPs was -94 mV, suggesting that it was mediated by an increase in K+ conductance. Extracellular application of tetraethylammonium or apamin, or intracellular injection of Cs+ or the Ca2+ chelating agent EGTA, blocked the Ca2+ spike AHP, indicating that it is mediated by a Ca(2+)-activated K+ current. 4. Block of the AHP resulted in the marked enhancement of a slow after-depolarizing potential (ADP). The slow ADP occurred only following the generation of low threshold Ca2+ spikes. Replacement of extracellular Ca2+ with Mg2+ or Sr2+ resulted in an abolition of the slow ADP. In addition, the increase in [Mg2+]o resulted in an abolition of the low threshold Ca2+ spike. In contrast, replacement of extracellular Ca2+ with Ba2+ did not abolish the slow ADP. These results indicate that the ADP can be activated by either Ca2+ or Ba2+, but not by Mg2+ or Sr2+. 5. Replacement of extracellular Na+ with choline+ did not abolish the slow ADP, while replacement with N-methyl-D-glucamine+ did, indicating that the slow ADP can be supported by choline+, but not by N-methyl-D-glucamine+. Neither chemical affected the low threshold Ca2+ spike. These results are consistent with the slow ADP being mediated by a Ca(2+)-activated non-selective cation (CAN) current.(ABSTRACT TRUNCATED AT 400 WORDS)

Halothane-induced hyperpolarization and depression of postsynaptic potentials of guinea pig thalamic neurons in vitro

Intralaminar thalamic nuclei have been considered to be a component of the non-specific sensory system which is involved in physiological functions related to consciousness and pain sensation. The effect of halothane on membrane potentials and synaptic properties of neurons of the parafascicular (Pf) nucleus in guinea pig brain slices was investigated using intracellular recording methods. Halothane at concentrations of 0.4–1.0 mM, which are in the range of clinical concentrations, produced hyperpolarizations of 2–8 mV in approximately 50% of the cells. The halothane-induced hyperpolarization was nullified at a membrane potential close to the K + equilibrium potential. The amplitude of the hyperpolarization was dependent on the external K + concentration, and was decreased by either Ba 2+, or 4-aminopyridine, or intracellular injection of Cs +. All these results indicate that the hyperpolarization was due to an increase in K + conductance. Halothane at clinical concentrations depressed both excitatory and inhibitory postsynaptic potentials in a concentration-dependent manner. On the other hand hyperpolarizing responses to exogenous γ-aminobutyric acid (GABA) in the presence of bicuculline were suppressed by halothane, but depolarizing responses to l-glutamate were not altered. The results indicate that the depressant action of the anesthetic on the excitatory postsynaptic potential (EPSP) may occur presynaptically, whereas the blocking action on the inhibitory postsynaptic potential (IPSP) may occur postsynaptically.

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

1. To study the synaptic mechanisms of excitatory transmission in the suprachiasmatic nucleus (SCN), we assessed the effects of excitatory amino acid receptor antagonists on excitatory postsynaptic potentials (EPSPs) recorded from SCN neurons in horizontal and parasagittal hypothalamic slice preparations from rats and guinea-pigs. The EPSPs were evoked by electrical stimulation of either optic nerve or a site near the SCN. 2. When evoked at membrane potentials between -60 and -100 mV, the EPSPs from optic nerve stimulation were conventional in shape; they rose to the peak quickly (6.2 +/- 0.5 ms, mean +/- S.E.M.; n = 45) and decayed gradually over 50-250 ms. When evoked at membrane potentials between -20 and -55 mV after blockade of outward K+ currents and fast Na+ spikes by intracellular injection of Cs+ and QX-314 (n = 5 neurons), a slow depolarizing potential emerged near the fast peak of the EPSP. This slow potential, unlike the fast peak, was not linearly related to membrane potential. 3. An antagonist for kainate- and quisqualate-type excitatory amino acid receptors, 6,7-dinitroquinoxaline-2,3-dione (DNQX 1-10 microM), depressed in a concentration-dependent and reversible manner the EPSPs evoked by optic nerve stimulation at membrane potentials between -60 and -100 mV (n = 9). The effects of DNQX were not associated with any significant changes in the baseline input resistance or membrane potential of the postsynaptic neurons. The selective N-methyl-D-aspartate (NMDA) receptor antagonist, DL-2-amino-5-phosphonopentanoic acid (AP5, 50-100 microM), did not affect significantly and consistently the EPSPs evoked at these membrane potentials (n = 7). On the other hand, AP5 (50 microM) blocked or depressed the slow depolarizing component of the EPSPs evoked at membrane potentials between -20 and -55 mV (n = 4). No significant changes in baseline input resistance or membrane potential accompanied the effects of AP5. 4. Stimulation of a site lateral or dorsocaudal to the SCN evoked EPSPs distinct from those evoked by optic nerve stimulation. Again, DNQX (0.3-10 microM) depressed the EPSPs evoked at membrane potentials between -60 and -100 mV (n = 4) whereas AP5 (50 microM) had no effect (n = 5). When evoked at less negative membrane potentials (i.e. -20 to -55 mV) after intracellular injection of Cs+ and QX-314, the EPSPs had a slow depolarizing potential, similar to the EPSPs from optic nerve stimulation.(ABSTRACT TRUNCATED AT 400 WORDS)

Functional properties of a slowly inactivating potassium current in guinea pig dorsal lateral geniculate relay neurons

1. The time- and voltage-dependent properties of a slowly inactivating and depolarization-activated potassium current and the functional consequences of its activation was investigated with current and single-electrode voltage-clamp techniques applied to guinea pig dorsal lateral geniculate neurons maintained as a slice in vitro. 2. In current clamp, application of a step depolarization to near firing threshold resulted in a slowly rising membrane potential that took up to 10 s to reach steady state and firing threshold. In voltage clamp, step depolarization of the membrane potential to values positive to approximately -65 mV resulted in the rapid activation followed by slow inactivation of an outward current. In both cases the sudden depolarization was associated with a large increase in membrane conductance, which gradually lessened in parallel with the slow depolarization in current clamp or with the decrease in outward current in voltage clamp. 3. The time course of inactivation of the outward current, which we refer to as IAs, was well fitted by a two-exponential function with time constants of 96 and 2,255 ms, suggesting the presence of a fast and slow phase of inactivation. The activation threshold for IAs was about -65 to -60 mV, whereas inactivation was incomplete even at -50 mV, suggesting the presence of a substantial "window" current. The time course of removal of inactivation of IAs at -85 to -100 mV was well fitted by a single exponential function with time constant of 91 ms. 4. IAs appears to be mediated by K+. Increasing [K+]o from 2.5 to 10 mM resulted in a reduction in amplitude of IAs, whereas changing from 10 to 2.5 mM [K+]o enhanced this current. Intracellular injection of Cs+ resulted in an abolition of IAs, whereas extracellular application of Ba2+ resulted in a large decrease in the apparent input conductance but relatively little reduction of IAs. 5. Both phases of inactivation of the transient outward current were completely blocked by low doses (100 microM) of 4-aminopyridine (4-AP), but not by extracellular application of Cs+, tetraethylammonium (TEA), tetrodotoxin (TTX), or after block of transmembrane Ca2+ currents. Local application of 4-AP to neurons depolarized to near firing threshold resulted in depolarization associated with a decrease in apparent input conductance, thereby confirming the presence of a window current.4+ this bias against depolarizing inputs.(ABSTRACT TRUNCATED AT 400 WORDS)

Actions of acetylcholine in the guinea-pig and cat medial and lateral geniculate nuclei, in vitro.

1. The mechanisms of action of acetylcholine (ACh) in the medial (m.g.n.) and dorsal lateral geniculate (l.g.n.d.) nuclei were investigated using intracellular recordings techniques in guinea-pig and cat in vitro thalamic slices. 2. Application of ACh to neurones in guinea-pig geniculate nuclei resulted in a hyperpolarization in all neurones followed by a slow depolarization in 52% of l.g.n.d. and 46% of m.g.n. neurones. Neither the hyperpolarization nor the slow depolarization were eliminated by blockade of synaptic transmission and both were activated by acetyl-beta-methylcholine and DL-muscarine and blocked by scopolamine, indicating that these responses are mediated by direct activation of muscarinic receptors on the cells studied. 3. The ACh-induced hyperpolarization was associated with an increase in apparent input conductance (Gi) of 4-13 nS. The reversal potential of the ACh-induced hyperpolarization varied in a Nernstian manner with changes in extracellular [K+] and was greatly reduced by bath application of the K+ antagonist Ba2+ or intracellular injection of Cs+. These findings show that the muscarinic hyperpolarization is mediated by an increase in K+ conductance. 4. The ACh-induced slow depolarization was associated with a decrease in Gi of 2-15 nS, had an extrapolated reversal potential near EK, and was sensitive to [K+]o, indicating that this response is due to a decrease in K+ conductance. 5. In contrast to effects on guinea-pig geniculate neurones, applications of ACh to cat l.g.n.d. and m.g.n. cells resulted in a rapid depolarization in nearly all cells, followed in some neurones by a hyperpolarization and/or a slow depolarization. The rapid excitatory response was associated with an increase in membrane conductance, had an estimated reversal potential of -49 to -4 mV and may be mediated by nicotinic receptors. The hyperpolarization and slow depolarization were similar to those of the guinea-pig in that they were associated with an increase and decrease, respectively, of Gi, and were mediated by muscarinic receptors. 6. The muscarinic hyperpolarization interacted with the intrinsic properties of the thalamic neurones to inhibit single-spike activity while promoting the occurrence of burst discharges. The muscarinic slow depolarization had the opposite effect; it brought the membrane potential into the range where burst firing was blocked and single-spike firing predominated. Depending upon the membrane potential, the rapid excitatory response of cat geniculate neurones could activate either a burst or a train of action potentials.(ABSTRACT TRUNCATED AT 400 WORDS)

Pharmacological evidence for L-aspartate as the neurotransmitter of cerebellar climbing fibres in the guinea-pig.

Climbing fibre responses (c.f.r.s) evoked by white matter stimulation and the depolarizations induced by iontophoretically applied L-glutamate and L-aspartate were recorded intracellularly from the proximal dendrites of Purkinje cells in in vitro slice preparations of the guinea-pig cerebellum. Short pulses of L-glutamate and L-aspartate dose-dependently depolarized the Purkinje cell dendrite. Even small doses of these amino acids reduced the input resistance. The maximum decrease in input resistance induced by L-glutamate was 36% and that by L-aspartate was 38%. Intracellular injection of Cs+ allowed Purkinje cell dendrites to be depolarized to a range of -15 to +30 mV. The mean reversal potential for the c.f.r. (Ec) was found to be +10.2 mV (n = 4). The mean reversal potentials obtained for L-glutamate (Eg) and for L-aspartate (Ea) were +7.3 mV (n = 7) and +5.6 mV (n = 7) respectively. When external Na+ concentration was reduced, Ec, Ea and Eg were linearly and similarly shifted in the negative direction, indicating that all these reversal potentials are determined primarily by a Na+ conductance. The effects of the glutamate antagonists 2-amino-5-phosphonovaleric acid (APV), gamma-D-glutamylglycine (gamma-DGG), N-methyl-DL-aspartic acid (NMDLA) and glutamic acid diethylester (GDEE) were compared as to the responses to L-glutamate and L-aspartate and Ca2+-activated focal climbing fibre responses (c.f.c.f.r.s) in order to investigate the receptor type at the synapses formed by the climbing fibres with Purkinje cell dendrites. The order of antagonistic potency to the c.f.c.f.r. was : APV (mean percentage blockade = 99%) greater than gamma-DGG (87%) greater than NMDLA (71%) greater than GDEE (28%). The order of antagonistic potency to the response to L-aspartate was: gamma-DGG (69%) greater than APV (66%) greater than NMDLA (60%) greater than GDEE (31%), and that to the response to L-glutamate was: GDEE (63%) greater than NMDLA (22%) greater than gamma-GDD (15%) greater than APV (14%). APV was found to be the most effective anatagonist of the c.f.c.f.r. Its action was reversible, selective for L-aspartate-induced depolarization and had no effect on the responses to L-glutamate. NMDLA, which has no activity as an agonist, was a greater suppressant of the responses to L-aspartate than those to L-glutamate. These electrophysiological and pharmacological findings suggest that the receptor for the transmitter at the synapses formed by climbing fibres with Purkinje cell dendrites is of the L-aspartate-preferring type, and are thus consistent with the bio-and histochemical findings that L-aspartate may be the endogenous transmitter at this synapse.

The reversal potential of excitatory amino acid action on granule cells of the rat dentate gyrus.

The responses of granule cells to glutamate, aspartate, N-methyl-D-aspartate (NMDA), quisqualate and kainate applied by ionophoresis on to their dendrites in the middle molecular layer of the dentate gyrus were studied with intracellular electrodes using an in vitro hippocampal slice preparation. On passive depolarization 75% of the granule cells displayed anomalous rectification, which persisted in the presence of TTX and TEA but was eliminated by Co2+ or the intracellular injection of Cs+. Short ionophoretic applications of all the excitatory amino acids evoked dose-dependent depolarizations that were highly localized: movement of the ionophoretic electrode by as little as 10 microns could substantially change the size of the response. The depolarizations evoked by glutamate, asparatate, quisqualate and kainate were unaffected by TTX and Co2+. The depolarization evoked by NMDA was unaffected by TTX but markedly reduced by Co2+. Following intracellular injection of Cs+, neurones could be depolarized to +30 mV and the depolarizations produced by glutamate, quisqualate, NMDA and kainate reversed. The reversal potentials (E) were Eglutamate: -5.6 +/- 0.4 mV; ENMDA: 1.8 +/- 1.9 mV; Equisqualate: -3.9 +/- 1.9 mV; Ekainate: -4.6 +/- 2.0 mV. The excitatory post-synaptic potential (e.p.s.p.) evoked by stimulation of the medial perforant path could also be reversed and Ee.p.s.p. was -5.5 +/- 1.1 mV. The 6 mV difference between ENMDA and the equilibrium potential for the other exogenously applied excitatory amino acids and the statistically significant difference between ENMDA and Ee.p.s.p. (P less than 0.005; d.f.: 7) is consistent with our earlier hypothesis that both the transmitter released by the medial perforant path and exogenously applied glutamate are unlikely to interact with NMDA receptors.

Coexistence of GABAA and GABAB receptors on A delta and C primary afferents.

Intracellular recordings from adult rat dorsal root ganglion neurones were performed in vitro and the coexistence of two gamma-aminobutyric acid (GABA) receptors on the membrane of identified A delta and C primary afferents was demonstrated. Transient applications of GABA (10(-6)-10(-2) M) evoked dose-dependent depolarizations and increased membrane conductance. The responses were mimicked by muscimol, isoguvacine, THIP and 3 amino propane sulphonic acid (3 APS); they were blocked by bicuculline and picrotoxin. Pentobarbitone induced an increase of GABA-induced depolarizations. Perfusion of tetraethylammonium (TEA, 7.5 mM) and intracellular injection of Cs+ ions unmasked the Ca2+ component of action potentials, which appeared as long-lasting plateau depolarizations. Such action potentials were shortened in the presence of methoxyverapamil (D600, 5 X 10(-6)-10(-5) M) and in a medium without Ca+ ions. Prolonged (5-10 min) perfusion of GABA (10(-9)-10(-5) M) shortened the Ca2+ component of action potentials. This effect was mimicked by baclofen (10(-7)-5 X 10(-6) M) and muscimol (5 X 10(-7)-10(-5) M) and was not affected by bicuculline perfusion (5 X 10(-6)-10(-5) M). Isoguvacine (2.5 X 10(-5) M) did not affect action potential duration. It is concluded that two GABA receptors coexist on the membrane of slow conducting primary afferents: the bicuculline-sensitive GABAA receptor mediates depolarizations and the bicuculline-insensitive GABAB receptor shortens the calcium component of action potentials.

Excitation of hippocampal pyramidal cells by glutamate in the guinea-pig and rat.

1. The mechanism by which L-glutamic acid depolarizes hippocampal CA1 pyramidal neurones was investigated by using the in vitro slice and ionophoretic techniques. 2. Two types of responses were seen. One (in 85% of cells) consisted of spike discharges that outlasted the glutamate-induced depolarization. In the other (the rest of the cells), spikes were produced only during the rising phase of the depolarization. 3. The effect was highly localized; it disappeared when the ionophoretic electrode was moved vertically by as little as 20 micrometers. 4. The effect of glutamate persisted after synaptic transmission was blocked; this probably was due to a direct effect of glutamate on the cell membrane. 5. Small doses of glutamate produced either no change or an apparent increase in input resistance. With larger doses, the input resistance invariably decreased. The apparent increase in input resistance was not seen in cells treated with Mn2+ and TTX and is believed to be an effect of the depolarization rather than a direct effect of glutamate. 6. By extrapolation, the reversal potential for the glutamate response (EGlu) was found to -3.6 mV. 7. Following intracellular injection of Cs+, neurones could be depolarized to a range of +20 to +50 mV. The glutamate response could then be reversed. EGlu in these cells was -1.5 mV. 8. Using the Cs+-injection technique, it was also possible to reverse the e.p.s.p. E.e.p.s.p. was similar to EGlu. 9. When the external sodium concentration was reduced, the size of the glutamate response decreased, and EGlu became more negative.