Average Reversal Potential Research Articles

Catecholamines modulate the delayed rectifying potassium current (IK) in guinea pig ventricular myocytes

The effects of isoproterenol and norepinephrine on the delayed, outward potassium current, IK, were tested in single, dialyzed guinea pig heart cells where complications from a restricted extracellular space are minimized and internal K concentration is controlled. The average IK reversal potential in control cells was -80 +/- 2.1 mV (N = 4; [K]o = 4.5 mM; [K]i = 120 mM) indicating a high degree of K selectivity (PNa/PK less than or equal to 0.01). In paired experiments, both isoproterenol and norepinephrine increased IK without changing the reversal potential, indicating that the selectivity of the channels is not altered by these agents. The results explain the shortening of action potential duration observed at high concentrations (Quadbeck and Reiter, 1975); and suggest that an increase in IK by catecholamines may serve to limit the degree of calcium current-induced action potential prolongation during increased sympathetic tone and rapid heart rates.

Electrophysiologic effects of nicorandil, a new antianginal agent, on action potentials and membrane currents of rabbit atrioventricular node.

Electrophysiologic effects of nicorandil, a newly developed coronary vasodilator, on the atrioventricular (AV) node were studied using space-clamped small preparations of the rabbit AV node. At the concentrations of 10(-6), 10(-5) and 10(-4) mol/l, this drug did not cause significant changes in the action potential characteristics including the spontaneous firing frequency, overshoot, maximum diastolic potential, maximum rate of depolarization and action potential duration. When the resting membrane potential of the AV node was obtained by a superfusion with verapamil and then nicorandil was added to the perfusate, no hyperpolarization was observed. This was in sharp contrast to the reported hyperpolarizing action of this drug on the atrial muscle, Purkinje fibres and vascular smooth muscle where the resting potential is much more negative. On the other hand, voltage clamp experiments using double microelectrode techniques revealed that 10(-5) to 10(-4) mol/l nicorandil increased the steady-state outward current at potentials positive to -40 mV and the steady-state inward current at potentials negative to -40 mV. Such a nicorandil-sensitive component of the steady-state current had an average reversal potential of -41.8 mV (n = 5). This component was considered to reflect changes in the time-independent background current, although, at more negative potential levels, it may partially reflect an increase in the hyperpolarization activated inward current (ih). Nicorandil, at the concentrations of 10(-5) and 10(-4) mol/l, increased the slow inward current (isi) by 10.8% (n = 5, p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine causes rapid nicotinic excitation in the medial habenular nucleus of guinea pig, in vitro

The actions of ACh in the medial habenular nucleus (MHb) were investigated using extra- and intracellular recording techniques in guinea pig thalamic slice maintained in vitro. Applications of ACh to MHb neurons resulted in rapid excitation followed by inhibition. Neither of these responses was abolished by blockade of synaptic transmission, indicating that they are consequences of ACh action directly on MHb cells. Local applications of the nicotinic agonists nicotine and cytisine caused long-lasting excitation, while applications of another nicotinic agonist, 1,1-dimethyl-4-phenylpiperazinium caused both the excitatory and inhibitory responses. Applications of the muscarinic agonists DL-muscarine and acetyl-beta-methylcholine did not consistently cause either the excitatory or inhibitory response. Adding the nicotinic antagonist hexamethonium to the bathing medium blocked both the excitatory and inhibitory ACh responses, while addition of the muscarinic antagonists atropine or scopolamine had no effect. These results indicate that the effects of ACh on MHb neurons are mediated by nicotinic receptors. Intracellular recordings revealed that ACh or nicotine cause an increase in membrane conductance associated with depolarizations that had an average reversal potential of -16 to -11 mV. These results indicate that the ACh-induced excitation is due to an increase in membrane cation conductance. The inhibitory response that follows ACh-induced depolarization and repetitive firing was associated with a hyperpolarization and an increase in membrane conductance. Similar postexcitatory inhibition could also be elicited by direct depolarization or by applications of glutamate, indicating that the hyperpolarizing response to ACh may be an endogenous postexcitatory potential that is not directly coupled to activation of nicotinic receptors. These results suggest that cholinergic transmission in the MHb may be largely of the nicotinic type. This nucleus may be of one of the major regions of the nervous system through which nicotine mediates its central effects.

Mechanisms of action of acetylcholine in the guinea-pig cerebral cortex in vitro.

The mechanisms of action of acetylcholine (ACh) in the guinea-pig neocortex were investigated using intracellular recordings from layer V pyramidal cells of the anterior cingulate cortical slice. At resting membrane potential (Vm = -80 to -70 mV), ACh application resulted in a barrage of excitatory and inhibitory post-synaptic potentials (p.s.p.s) associated with a decrease in apparent input resistance (Ri). ACh, applied to pyramidal neurones depolarized to just below firing threshold (Vm = -65 to -55 mV), produced a short-latency hyperpolarization concomitant with p.s.p.s and a decrease in Ri, followed by a long-lasting (10 to greater than 60 s) depolarization and action potential generation. Both of these responses were also found in presumed pyramidal neurones of other cortical regions (sensorimotor and visual) and were blocked by muscarinic, but not nicotinic, antagonists. The ACh-induced hyperpolarization possessed an average reversal potential of -75.8 mV, similar to that for the hyperpolarizing response to gamma-aminobutyric acid (GABA; -72.4 mV) and for the i.p.s.p. generated by orthodromic stimulation (-69.6 mV). This cholinergic inhibitory response could be elicited by ACh applications at significantly greater distance from the cell than the slow depolarizing response. Blockade of GABAergic synaptic transmission with solution containing Mn2+ and low Ca2+, or by local application of tetrodotoxin (TTX), bicuculline or picrotoxin, abolished the ACh-induced inhibitory response but not the slow excitatory response. In TTX (or Mn2+, low Ca2+) the slow excitatory response possessed a minimum onset latency of 250 ms and was associated with a voltage-dependent increase in Ri. Application of ACh caused short-latency excitation associated with a decrease in Ri in eight neurones. The time course of this excitation was similar to that of the inhibition seen in pyramidal neurones. Seven of these neurones had action potentials with unusually brief durations, indicating that they were probably non-pyramidal cells. ACh blocked the slow after-hyperpolarization (a.h.p.) following a train of action potentials, occasionally reduced orthodromically evoked p.s.p.s, and had no effect on the width or maximum rate of rise or fall of the action potential. It is concluded that cholinergic inhibition of pyramidal neurones is mediated through a rapid muscarinic excitation of non-pyramidal cells, resulting in the release of GABA. In pyramidal cells ACh causes a relatively slow blockade of both a voltage-dependent hyperpolarizing conductance (M-current) which is most active at depolarized membrane potentials, and the Ca2+-activated K+ conductance underlying the a.h.p.(ABSTRACT TRUNCATED AT 400 WORDS)

Synaptic interactions between mammalian central neurons in cell culture. I. Reversal potential for excitatory postsynaptic potentials.

Intracellular recording and stimulation techniques were used to study the electrical properties of neurons in cell cultures from fetal mouse spinal cord (SC). The morphology of SC neurons and the distribution on SC neurons of boutons formed by synaptically connected SC or dorsal root ganglion (DRG) neurons were demonstrated with horseradish peroxidase (HRP) injection. Postsynaptic polarization in conjunction with synaptic activation of SC neurons was used to determine the reversal potential for excitatory postsynaptic potentials (EPSPs). Tetraethylammonium ions were injected postsynaptically in order to obtain reversal of the EPSPs. Both SC-SC and DRG-SC excitatory connections could be reversed by postsynaptic depolarization. The average reversal potential for the SC-SC EPSP was -4 +/- 12.2 (SD) mV and that for the DRG-SC EPSP was +8 +/- 7.9 (SD) mV, a statistically significant difference (Wilcoxon two-sample rank; P less than 0.05). Scatter was quite large, particularly for the SC-SC connection. While some neurons gave clear electrophysiological evidence of significant dendritic effects, the average total electrotonic length was small (0.58 +/- 0.65 (SD) of a length constant). The morphological extent of the dendrites of SC neurons was substantially less than that of mature motoneurons in vivo. We concluded that both SC-SC and DRG-SC EPSPs were mediated by a conventional conductance increase and that most synaptic input was not far removed electrically from the recording site in the neuron cell body.

A depolarizing inhibitory potential in neurones of the olfactory cortex in vitro.

1. Stable intracellular recordings were obtained from neurones in slices of the guinea-pig olfactory cortex maintained in vitro. 2. Single stimuli applied to the lateral olfactory tract (l.o.t.) produced an excitatory post-synaptic potential (e.p.s.p.) usually generating a single spike. 3. The e.p.s.p. was followed by a long (200-500 msec) after-depolarization (l.a.d.) of peak amplitude 5-16 mV. This was accompanied by a very large conductance increase and was associated with an inhibition of the intracellularly recorded e.p.s.p. and of spike generation. 4. The l.a.d. was more susceptible than the e.p.s.p. to depression by (i) repetitive l.o.t. stimulation and (ii) raising external [Mg2+]. The l.a.d. could be generated without a preceding spike. 5. At an average resting membrane potential of -74 mV the average reversal potential for the l.a.d. (El.a.d.) was -63 mV.El.a.d. became more positive on reducing [Cl-]out or on using KCl-filled electrodes. 6. It is concluded that the l.a.d. represents a Cl- -mediated inhibitory post-synaptic potential, generated through deep-lying recurrent inhibitory loops.

The permeability of aconitine-modified sodium channels to univalent cations in myelinated nerve

1. 1. Ionic currents through the sodium system of nodes of Ranvier treated with aconitine were measured under voltage clamp conditions in a Ringer solution containing Na + or an equimolar amount of various test cations. 2. 2. Average shifts in reversal potentials in nodes of Ranvier treated with aconitine with NH 4 +, Li +, K +, Rb +, Cs + in place of Na + in the Ringer solution are 7.6, −6.8, −25.0, −41.0 and −51.5 mV at 13–14°C. At 20–22°C the sequence of shifts is 7.5, −5.5, −13.5, −29.0 and −41.0 mV. For Tl + the average reversal potential shift is +3 mV at 20–22°C. 3. 3. The slope of the instantaneous current-voltage relation at the reversal potential in nodes treated with aconitine changed with the various cations tested. The ratios are NH 4 +/Na +/K +/Rb +/Cs +/Li + = 1.14 : 1.0 : 0.80 : 0.67 : 0.53 : 0.53. 4. 4. Using a three energy barrier model some of the parameters for the aconitine-modified Na + channels were estimated (Chizmadgev, Yu. A., Khodorov, B.I. and Aityan, S.Kh. (1974) Bioelectrochem. Bioenerg. 1, 301–312).