Canadian Journal of Physiology and Pharmacology, 1992
Slow excitatory postsynaptic potentials in sympathetic ganglia often involve suppression of a vol... more Slow excitatory postsynaptic potentials in sympathetic ganglia often involve suppression of a voltage-dependent potassium current termed the M current. This current is suppressed by the muscarinic action of acetylcholine, by peptides such as luteinizing hormone releasing hormone, and sometimes by α-adrenoceptor agonists. Activation of β-adrenoceptors sometimes produces weak potentiation. The voltage dependence of the M current is such that its suppression increases the excitability of ganglionic neurones. Since this sometimes leads to spontaneous discharge, activation of the slow excitatory postsynaptic potential mechanism (or modulation of M current) within a sympathetic ganglion produces effects that manifest in the autonomic outflow to the target organ. In frogs, M currents are present in the neurones of both paravertebral sympathetic ganglia and cardiac parasympathetic ganglia. Since the M current is suppressed by adrenaline in the parasympathetic ganglia and these ganglia often...
Journal of Pharmacology and Experimental Therapeutics
The food dye erythrosin B has been reported to inhibit the neuronal uptake of catecholamines. To ... more The food dye erythrosin B has been reported to inhibit the neuronal uptake of catecholamines. To test this hypothesis we examined the effect of the dye on the responses of neurons in amphibian sympathetic ganglia to both dopamine and epinephrine. Although the hyperpolarizations induced by both catecholamines were potentiated by the conventional uptake blocker, desipramine (0.5 microM), low doses of erythrosin B (1-10 microM) produced an irreversible blockade. It was therefore not possible to evaluate the hypothesis that the dye might block catecholamine uptake. Xanthine dyes such as erythrosin B have also been reported to inhibit Na-K-adenosine triphosphatase. In agreement with this possibility we found that erythrosin B promoted irreversible inhibition of the (nicotinic) acetylcholine after-hyperpolarization. This response is generated by the electrogenic activity of the Na+ pump. Neither this Na+ pump inhibition nor erythrosin-induced membrane hyperpolarization appeared to account for the effect of the dye on catecholamine responses. The irreversible antagonism of epinephrine and dopamine by erythrosin was specific in that hyperpolarizing responses to muscarinic antagonists such as methacholine were relatively insensitive to the dye. It is therefore concluded that erythrosin B selectively antagonizes responses to catecholamines in amphibian sympathetic ganglia. No information as to the exact molecular mechanism of this antagonism is available from the present experiments.
The adrenaline-induced hyperpolarization (AdH) and the responses evoked by muscarine and luteiniz... more The adrenaline-induced hyperpolarization (AdH) and the responses evoked by muscarine and luteinizing hormone releasing hormone (LHRH) were recorded from neurones in amphibian sympathetic ganglia by means of the sucrose gap technique. The amplitude of the AdH was reduced when 'M-channel' closure was promoted by superfusion of LHRH or muscarine. 4-Aminopyridine (4-AP, 1 mM) antagonized the AdH, but not the depolarization evoked by muscarinic agonists. This implies that the channels involved in the electrogenesis of the AdH have different pharmacological properties from 'M-channels' and that the AdH is not generated by the opening of 'M-channels' outside their normal voltage range. Possible explanations for the attenuation of the AdH by muscarine and LHRH might be that (i) intracellular biochemical changes produced by these substances somehow interfere with the generation of the AdH or that (ii) muscarine and LHRH have allosteric interactions with the adrenoceptor mediating the AdH.
The pontine parabrachial nucleus (PBN) receives both opioid and Neuropeptide FF (NPFF) projection... more The pontine parabrachial nucleus (PBN) receives both opioid and Neuropeptide FF (NPFF) projections from the lower brain stem and/or the spinal cord. Because of this anatomical convergence and previous evidence that NPFF displays both pro- and anti-opioid activities, this study examined the synaptic effects of NPFF in the PBN and the mechanisms underlying these effects using an in vitro brain slice preparation and the nystatin-perforated patch-clamp recording technique. Under voltage-clamp conditions, NPFF reversibly reduced the evoked excitatory postsynaptic currents (EPSCs) in a dose-dependent fashion. This effect was not accompanied by apparent changes in the holding current, the current-voltage relationship or alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-induced inward currents in the PBN cells. When a paired-pulse protocol was used, NPFF increased the ratio of these synaptic currents. Analysis of miniature EPSCs showed that NPFF caused a rightward shift in the freque...
The present study examined the electrophysiological and kinetic properties of a hyperpolarizing-a... more The present study examined the electrophysiological and kinetic properties of a hyperpolarizing-activated current in neurons located in the lateral parabrachial nucleus. We investigated whether differences observed in the shape of action potential afterhyperpolarizations in lateral parabrachial nucleus neurons, and the ability of these neurons to accommodate, correlated with the presence of this current. A voltage-activated inwardly rectifying current that increased in amplitude with hyperpolarization was observed in 83% of the neurons examined. Under voltage-clamp recording conditions, this current activated at about -70 mV, was half-activated at -96.5 mV and was blocked by bath application of 2 mM cesium, but not by 100 microM barium. In the current-clamp mode, activation of this current resulted in a transient increase in neuronal excitability at the termination of the more negative current injections. The presence of this current did not correlate with specific action potential ...
1. Neurones dissociated from Rana pipiens paravertebral sympathetic ganglia were studied by means... more 1. Neurones dissociated from Rana pipiens paravertebral sympathetic ganglia were studied by means of the whole-cell patch-clamp technique. Responses to agonists were best recorded when cyclic AMP was included in the patch pipette. 2. Two populations of cells were identified on the basis of size (input capacitance, Cin) and the presence or absence of a fast, transient outward current (A-current, IA). This current was usually present in the 'large' cells (Cin = 40.5 +/- 1.5 pF, n = 66) but absent from 'small' cells (Cin = 21.0 +/- 0.8 pF, n = 70). 3. Both cell types exhibited a slowly activating, non-inactivating K+ current (M-current, IM) which was suppressed by luteinizing hormone-releasing hormone (LHRH, 10-100 microM). Threshold for activation of IM was about -75 mV, half-maximal activation was at -50 mV and the M-conductance GM increased e-fold for at 7 mV change in membrane potential. The maximum value for IM studied in large cells by patch-clamp procedures was l...
The Journal of neuroscience : the official journal of the Society for Neuroscience, 1991
Although dopamine (DA)-containing neurons participate in a number of important cerebral functions... more Although dopamine (DA)-containing neurons participate in a number of important cerebral functions, the physiology of their synaptic connections is poorly understood. By using whole-cell patch-clamp recording in thin slices of rat mesencephalon, we have investigated the biophysical properties of synaptic events and the nature of neurotransmitter(s) and receptors involved in the synaptic input to DA neurons in substantia nigra. The histological and electrophysiological characteristics of these cells were consistent with those described by recent in vivo and in vitro studies, thus allowing their unequivocal identification. Under appropriate experimental conditions, intranigral stimulation produced excitatory synaptic inputs in DA neurons. By voltage-clamp analysis, most of these excitatory postsynaptic currents (EPSCs) had a rise time of about 1.0 msec and a decay phase that could be fit by the sum of two exponential curves so that a fast and a slow component could be distinguished. Th...
1. The whole-cell patch-clamp technique was used to record membrane currents from neurones which ... more 1. The whole-cell patch-clamp technique was used to record membrane currents from neurones which were acutely dissociated from the intra-atrial parasympathetic ganglia of Rana pipiens. The effects of muscarine and adrenaline were observed at a holding potential of -30 mV. Extracellular potassium concentration ([K+]o) was 2, 6 or 20 mM. 2. Muscarine (10 microM) produced inward current in thirteen cells, outward current in eighteen cells and seven cells were unaffected. Inward currents were observed in six out of ten neurones in which the intracellular solution contained adenosine triphosphate (ATP; 100 microM) and outward currents were seen in eleven out of fourteen neurones which contained adenosine 3',5'-cyclic monophosphate (cyclic & 100 microM). 3. In five out of nine cells tested, the inward current produced by muscarine was attributable to a 30% depression of a voltage-dependent current which resembled the M-current (IM). Muscarine-induced inward current in the other fo...
The adrenaline-induced hyperpolarization, which was recorded in neurons of bullfrog paravertebral... more The adrenaline-induced hyperpolarization, which was recorded in neurons of bullfrog paravertebral sympathetic ganglia by means of the sucrose gap technique, was antagonized by 1 mM 4-aminopyridine. The response was unaffected by drugs which influence intracellular Ca2+ movements or Ca2+-sensitive K+ conductances, i.e. 100 or 200 microM Cd2+, 60 microM dantrolene Na+, 10 mM tetraethylammonium bromide, 0.5-2.0 microM apamin or 70 microM (+)-tubocurarine chloride. The spontaneous, rhythmic hyperpolarizations which occur in ganglionic neurons in the presence of 5 mM caffeine and reflect activation of Ca2+-sensitive K+ conductances following mobilization of intracellular Ca2+, were examined by means of intracellular recording. These responses were often biphasic, comprising a transient rapid early phase and a slow late phase. Tetraethylammonium (10 mM) and 0.5-2.0 microM apamin antagonized the rapid early phase and 70 microM (+)-tubocurarine chloride antagonized both phases of the response. Neither phase of these spontaneous, rhythmic, caffeine-induced hyperpolarizations were affected by 1 mM 4-aminopyridine. Although the adrenaline-induced hyperpolarization was antagonized by 50 microM 8-(diethylamino)octyl-3,4,5-trimethoxybenzoate and by 50 microM quinidine, the majority of the results argue against the hypothesis that mobilization of intracellular Ca2+ is required for activation of the K+ conductance thought to underlie the adrenaline-induced hyperpolarization.
The potassium-activated hyperpolarization (KH) was used as an index of electrogenic Na+ pumping i... more The potassium-activated hyperpolarization (KH) was used as an index of electrogenic Na+ pumping in bullfrog sympathetic ganglia. This response was evoked by storing ganglia in K-free Ringer's solution and briefly introducing normal Ringer's solution containing 2 mM K+ at regular intervals. The apparent EC50 for K+ was 2.21 mM (range 0.88-3.54 mM, for n = 5) and at least 10 mM K+ was required to produce a maximal KH response. Adrenaline, which produces membrane hyperpolarization by increasing K+ conductance (gK), increased the amplitude of KH responses. When the K+ efflux accompanying the adrenaline-induced hyperpolarization (AdH) was blocked with 2 mM Ba2+, the KH was no longer potentiated. It is suggested that the K+ moving out of the cells during the AdH accumulates extracellularly and stimulates the Na+ pump.
Neuropeptide Y (NPY) is found in the C-cells yet is absent from the B-cells of amphibian sympathe... more Neuropeptide Y (NPY) is found in the C-cells yet is absent from the B-cells of amphibian sympathetic ganglia. The effects of this peptide on both cell types were studied using the whole-cell patch-clamp recording technique. Although NPY had little effect on the B-cells, it opened inwardly-rectifying K(+)-channels in C-cells. This effect of the peptide on C-cell K(+)-channels was similar to that produced by muscarine and adrenaline and may be related to autoreceptor function.
Canadian Journal of Physiology and Pharmacology, 1992
Slow excitatory postsynaptic potentials in sympathetic ganglia often involve suppression of a vol... more Slow excitatory postsynaptic potentials in sympathetic ganglia often involve suppression of a voltage-dependent potassium current termed the M current. This current is suppressed by the muscarinic action of acetylcholine, by peptides such as luteinizing hormone releasing hormone, and sometimes by α-adrenoceptor agonists. Activation of β-adrenoceptors sometimes produces weak potentiation. The voltage dependence of the M current is such that its suppression increases the excitability of ganglionic neurones. Since this sometimes leads to spontaneous discharge, activation of the slow excitatory postsynaptic potential mechanism (or modulation of M current) within a sympathetic ganglion produces effects that manifest in the autonomic outflow to the target organ. In frogs, M currents are present in the neurones of both paravertebral sympathetic ganglia and cardiac parasympathetic ganglia. Since the M current is suppressed by adrenaline in the parasympathetic ganglia and these ganglia often...
Journal of Pharmacology and Experimental Therapeutics
The food dye erythrosin B has been reported to inhibit the neuronal uptake of catecholamines. To ... more The food dye erythrosin B has been reported to inhibit the neuronal uptake of catecholamines. To test this hypothesis we examined the effect of the dye on the responses of neurons in amphibian sympathetic ganglia to both dopamine and epinephrine. Although the hyperpolarizations induced by both catecholamines were potentiated by the conventional uptake blocker, desipramine (0.5 microM), low doses of erythrosin B (1-10 microM) produced an irreversible blockade. It was therefore not possible to evaluate the hypothesis that the dye might block catecholamine uptake. Xanthine dyes such as erythrosin B have also been reported to inhibit Na-K-adenosine triphosphatase. In agreement with this possibility we found that erythrosin B promoted irreversible inhibition of the (nicotinic) acetylcholine after-hyperpolarization. This response is generated by the electrogenic activity of the Na+ pump. Neither this Na+ pump inhibition nor erythrosin-induced membrane hyperpolarization appeared to account for the effect of the dye on catecholamine responses. The irreversible antagonism of epinephrine and dopamine by erythrosin was specific in that hyperpolarizing responses to muscarinic antagonists such as methacholine were relatively insensitive to the dye. It is therefore concluded that erythrosin B selectively antagonizes responses to catecholamines in amphibian sympathetic ganglia. No information as to the exact molecular mechanism of this antagonism is available from the present experiments.
The adrenaline-induced hyperpolarization (AdH) and the responses evoked by muscarine and luteiniz... more The adrenaline-induced hyperpolarization (AdH) and the responses evoked by muscarine and luteinizing hormone releasing hormone (LHRH) were recorded from neurones in amphibian sympathetic ganglia by means of the sucrose gap technique. The amplitude of the AdH was reduced when 'M-channel' closure was promoted by superfusion of LHRH or muscarine. 4-Aminopyridine (4-AP, 1 mM) antagonized the AdH, but not the depolarization evoked by muscarinic agonists. This implies that the channels involved in the electrogenesis of the AdH have different pharmacological properties from 'M-channels' and that the AdH is not generated by the opening of 'M-channels' outside their normal voltage range. Possible explanations for the attenuation of the AdH by muscarine and LHRH might be that (i) intracellular biochemical changes produced by these substances somehow interfere with the generation of the AdH or that (ii) muscarine and LHRH have allosteric interactions with the adrenoceptor mediating the AdH.
The pontine parabrachial nucleus (PBN) receives both opioid and Neuropeptide FF (NPFF) projection... more The pontine parabrachial nucleus (PBN) receives both opioid and Neuropeptide FF (NPFF) projections from the lower brain stem and/or the spinal cord. Because of this anatomical convergence and previous evidence that NPFF displays both pro- and anti-opioid activities, this study examined the synaptic effects of NPFF in the PBN and the mechanisms underlying these effects using an in vitro brain slice preparation and the nystatin-perforated patch-clamp recording technique. Under voltage-clamp conditions, NPFF reversibly reduced the evoked excitatory postsynaptic currents (EPSCs) in a dose-dependent fashion. This effect was not accompanied by apparent changes in the holding current, the current-voltage relationship or alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-induced inward currents in the PBN cells. When a paired-pulse protocol was used, NPFF increased the ratio of these synaptic currents. Analysis of miniature EPSCs showed that NPFF caused a rightward shift in the freque...
The present study examined the electrophysiological and kinetic properties of a hyperpolarizing-a... more The present study examined the electrophysiological and kinetic properties of a hyperpolarizing-activated current in neurons located in the lateral parabrachial nucleus. We investigated whether differences observed in the shape of action potential afterhyperpolarizations in lateral parabrachial nucleus neurons, and the ability of these neurons to accommodate, correlated with the presence of this current. A voltage-activated inwardly rectifying current that increased in amplitude with hyperpolarization was observed in 83% of the neurons examined. Under voltage-clamp recording conditions, this current activated at about -70 mV, was half-activated at -96.5 mV and was blocked by bath application of 2 mM cesium, but not by 100 microM barium. In the current-clamp mode, activation of this current resulted in a transient increase in neuronal excitability at the termination of the more negative current injections. The presence of this current did not correlate with specific action potential ...
1. Neurones dissociated from Rana pipiens paravertebral sympathetic ganglia were studied by means... more 1. Neurones dissociated from Rana pipiens paravertebral sympathetic ganglia were studied by means of the whole-cell patch-clamp technique. Responses to agonists were best recorded when cyclic AMP was included in the patch pipette. 2. Two populations of cells were identified on the basis of size (input capacitance, Cin) and the presence or absence of a fast, transient outward current (A-current, IA). This current was usually present in the 'large' cells (Cin = 40.5 +/- 1.5 pF, n = 66) but absent from 'small' cells (Cin = 21.0 +/- 0.8 pF, n = 70). 3. Both cell types exhibited a slowly activating, non-inactivating K+ current (M-current, IM) which was suppressed by luteinizing hormone-releasing hormone (LHRH, 10-100 microM). Threshold for activation of IM was about -75 mV, half-maximal activation was at -50 mV and the M-conductance GM increased e-fold for at 7 mV change in membrane potential. The maximum value for IM studied in large cells by patch-clamp procedures was l...
The Journal of neuroscience : the official journal of the Society for Neuroscience, 1991
Although dopamine (DA)-containing neurons participate in a number of important cerebral functions... more Although dopamine (DA)-containing neurons participate in a number of important cerebral functions, the physiology of their synaptic connections is poorly understood. By using whole-cell patch-clamp recording in thin slices of rat mesencephalon, we have investigated the biophysical properties of synaptic events and the nature of neurotransmitter(s) and receptors involved in the synaptic input to DA neurons in substantia nigra. The histological and electrophysiological characteristics of these cells were consistent with those described by recent in vivo and in vitro studies, thus allowing their unequivocal identification. Under appropriate experimental conditions, intranigral stimulation produced excitatory synaptic inputs in DA neurons. By voltage-clamp analysis, most of these excitatory postsynaptic currents (EPSCs) had a rise time of about 1.0 msec and a decay phase that could be fit by the sum of two exponential curves so that a fast and a slow component could be distinguished. Th...
1. The whole-cell patch-clamp technique was used to record membrane currents from neurones which ... more 1. The whole-cell patch-clamp technique was used to record membrane currents from neurones which were acutely dissociated from the intra-atrial parasympathetic ganglia of Rana pipiens. The effects of muscarine and adrenaline were observed at a holding potential of -30 mV. Extracellular potassium concentration ([K+]o) was 2, 6 or 20 mM. 2. Muscarine (10 microM) produced inward current in thirteen cells, outward current in eighteen cells and seven cells were unaffected. Inward currents were observed in six out of ten neurones in which the intracellular solution contained adenosine triphosphate (ATP; 100 microM) and outward currents were seen in eleven out of fourteen neurones which contained adenosine 3',5'-cyclic monophosphate (cyclic & 100 microM). 3. In five out of nine cells tested, the inward current produced by muscarine was attributable to a 30% depression of a voltage-dependent current which resembled the M-current (IM). Muscarine-induced inward current in the other fo...
The adrenaline-induced hyperpolarization, which was recorded in neurons of bullfrog paravertebral... more The adrenaline-induced hyperpolarization, which was recorded in neurons of bullfrog paravertebral sympathetic ganglia by means of the sucrose gap technique, was antagonized by 1 mM 4-aminopyridine. The response was unaffected by drugs which influence intracellular Ca2+ movements or Ca2+-sensitive K+ conductances, i.e. 100 or 200 microM Cd2+, 60 microM dantrolene Na+, 10 mM tetraethylammonium bromide, 0.5-2.0 microM apamin or 70 microM (+)-tubocurarine chloride. The spontaneous, rhythmic hyperpolarizations which occur in ganglionic neurons in the presence of 5 mM caffeine and reflect activation of Ca2+-sensitive K+ conductances following mobilization of intracellular Ca2+, were examined by means of intracellular recording. These responses were often biphasic, comprising a transient rapid early phase and a slow late phase. Tetraethylammonium (10 mM) and 0.5-2.0 microM apamin antagonized the rapid early phase and 70 microM (+)-tubocurarine chloride antagonized both phases of the response. Neither phase of these spontaneous, rhythmic, caffeine-induced hyperpolarizations were affected by 1 mM 4-aminopyridine. Although the adrenaline-induced hyperpolarization was antagonized by 50 microM 8-(diethylamino)octyl-3,4,5-trimethoxybenzoate and by 50 microM quinidine, the majority of the results argue against the hypothesis that mobilization of intracellular Ca2+ is required for activation of the K+ conductance thought to underlie the adrenaline-induced hyperpolarization.
The potassium-activated hyperpolarization (KH) was used as an index of electrogenic Na+ pumping i... more The potassium-activated hyperpolarization (KH) was used as an index of electrogenic Na+ pumping in bullfrog sympathetic ganglia. This response was evoked by storing ganglia in K-free Ringer's solution and briefly introducing normal Ringer's solution containing 2 mM K+ at regular intervals. The apparent EC50 for K+ was 2.21 mM (range 0.88-3.54 mM, for n = 5) and at least 10 mM K+ was required to produce a maximal KH response. Adrenaline, which produces membrane hyperpolarization by increasing K+ conductance (gK), increased the amplitude of KH responses. When the K+ efflux accompanying the adrenaline-induced hyperpolarization (AdH) was blocked with 2 mM Ba2+, the KH was no longer potentiated. It is suggested that the K+ moving out of the cells during the AdH accumulates extracellularly and stimulates the Na+ pump.
Neuropeptide Y (NPY) is found in the C-cells yet is absent from the B-cells of amphibian sympathe... more Neuropeptide Y (NPY) is found in the C-cells yet is absent from the B-cells of amphibian sympathetic ganglia. The effects of this peptide on both cell types were studied using the whole-cell patch-clamp recording technique. Although NPY had little effect on the B-cells, it opened inwardly-rectifying K(+)-channels in C-cells. This effect of the peptide on C-cell K(+)-channels was similar to that produced by muscarine and adrenaline and may be related to autoreceptor function.
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Papers by J. Zidichouski