Basic Electrophysiological Properties Research Articles

17-Oestradiol modulates in vitro electrical properties and responses to kainate of oxytocin neurones in lactating rats.

1. Intracellular current clamp recordings were performed from identified oxytocin (OT) neurones in acute hypothalamic slices taken from lactating Wistar rats at early (5th day: LD-5) and late (21st day: LD-21) lactation. 2. The basic electrophysiological properties of LD-21 OT neurones differed from those of LD-5 OT neurones: their resting membrane potential was more depolarised (-51.5 versus -54.9 mV); their action potential duration was longer (1.6 versus 1.2 ms); their hyperpolarising after-potential (HAP) following single spikes and after-hyperpolarisation (AHP) following a burst of action potentials had smaller amplitudes (-46 and -67 %, respectively); and they lacked spike frequency adaptation during a burst. 3. In LD-21 neurones bath application of 17beta-oestradiol (10-7 M, 6-14 min) reversibly restored all these properties to values observed in LD-5 cells. This treatment had no effect on LD-5 neurones. 4. LD-21 neurones were less sensitive to kainate than LD-5 neurones. 17beta-Oestradiol significantly potentiated the kainate-induced response in LD-21, but not in LD-5 neurones. 5. The effects of 17beta-oestradiol were presumably mediated through a non-genomic mechanism since they occurred within a few minutes of administration, and disappeared within 30-40 min of washout. They were not inhibited by tamoxifen, an antagonist of the nuclear oestrogen receptor ER-alpha. Lastly, cholesterol, a non-active lipophilic molecule, had no effect. 6. Our observations demonstrate that, in the absence of 17beta-oestradiol, the basic electrical properties and sensitivity to kainate of OT neurones become altered between early and late lactation. However, the rise in circulating levels of oestrogens during the late phase of lactation may contribute to maintain OT neurone reactivity as long as suckling continues.

Intracellular recording and labeling of mossy cells and proximal CA3 pyramidal cells in macaque monkeys.

Little is known about the morphological characteristics and intracellular electrophysiological properties of neurons in the primate hippocampus and dentate gyrus. We have therefore begun a program of studies using intracellular recording and biocytin labeling in hippocampal slices from macaque monkeys. In the current study, we investigated mossy cells and proximal CA3 pyramidal cells. As in rats, macaque mossy cells display fundamentally different traits than proximal CA3 pyramidal cells. Interestingly, macaque mossy cells and CA3 pyramidal neurons display some morphological differences from those in rats. Macaque monkey mossy cells extend more dendrites into the molecular layer of the dentate gyrus, have more elaborate thorny excrescences on their proximal dendrites, and project more axon collaterals into the CA3 region. In macaques, three types of proximal CA3 pyramidal cells are found: classical pyramidal cells, neurons with their dendrites confined to the CA3 pyramidal cell layer, and a previously undescribed cell type, the "dentate" CA3 pyramidal cell, whose apical dendrites extend into and ramify within the hilus, granule cell layer, and molecular layer of the dentate gyrus. The basic electrophysiological properties of mossy cells and proximal CA3 cells are similar to those reported for the rodent. Mossy cells have a higher frequency of large amplitude spontaneous depolarizing postsynaptic potentials, and proximal CA3 pyramidal cells are more likely to discharge bursts of action potentials. Although mossy cells and CA3 pyramidal cells in macaque monkeys display many morphological and electrophysiological features described in rodents, these findings highlight significant species differences, with more heterogeneity and the potential for richer interconnections in the primate hippocampus.

Single streptomyces lividans K(+) channels: functional asymmetries and sidedness of proton activation.

Basic electrophysiological properties of the KcsA K(+) channel were examined in planar lipid bilayer membranes. The channel displays open-state rectification and weakly voltage-dependent gating. Tetraethylammonium blocking affinity depends on the side of the bilayer to which the blocker is added. Addition of Na(+) to the trans chamber causes block of open-channel current, while addition to the cis side has no effect. Most striking is the activation of KcsA by protons; channel activity is observed only when the trans bilayer chamber is at low pH. To ascertain which side of the channel faces which chamber, residues with structurally known locations were mapped to defined sides of the bilayer. Mutation of Y82, an external residue, results in changes in tetraethylammonium affinity exclusively from the cis side. Channels with cysteine residues substituted at externally exposed Y82 or internally exposed Q119 are functionally modified by methanethiosulfonate reagents from the cis or trans chambers, respectively. Block by charybdotoxin, known to bind to the channel's external mouth, is observed only when the toxin is added to the cis side of channels mutated to be toxin sensitive. These results demonstrate unambiguously that the protonation sites linked to gating are on the intracellular portion of the KcsA protein.

Ca channels in cardiac myocytes: structure and function in Ca influx and intracellular Ca release.

There are Ca channels in the plasma membrane and also the sarcoplasmic reticulum (SR) membrane in cardiac myocytes. The relationship between channel structure, associated proteins and function of these Ca channels is discussed. The sarcolemmal Ca channels are crucial both to the basic cellular electrophysiological properties and control of cardiac contractility (via excitation-contraction coupling). The intracellular Ca release channels (or ryanodine receptors) respond to triggering events mediated by sarcolemmal ion currents and are largely responsible for releasing Ca which activates the myofilaments to produce contraction. Several possible mechanisms of excitation-contraction coupling are discussed. The Ca released from the SR can also feedback on several sarcolemmal ion currents and alter action potential configuration as well as contribute to arrhythmogenesis.

Quantitative Structure-Activity Relationships of Antiarrhythmic Drugs

A Comprehensive review of quantitative structure-activity relationship (QSAR) studies on antiarrhythmic agents is presented. From the discussion point of view, the antiarrhythmic agents have been put into two broad classes: specific and nonspecific. While the main members of the former class can be beta-adrenergic blocking agents (beta-blockers), any chemical that can act directly on the myocardial cell membrane, producing a cardiodepressant effect via changes in basic electrophysiological properties of the membrane, such as automaticity, excitability, conductivity, and refractoriness, has been put in the latter class. QSARs exhibit that the biological actions of a variety of drugs belonging to any class depend primarily on the lipophilic haracter of the molecule or substituents. Thus, the hydrophobic interaction is found to play a dominant role in the action of antiarrhythmic agents. In certain cases, the QSARs also exhibit the role of electronic parameters, suggesting that certain receptors may have electronic site also, permitting the drugs to involve in some electronic interactions, too.

TNF alpha induces a protein kinase C-dependent reduction in astroglial K+ conductance.

Incubation of cultured cortical astrocytes with tumor necrosis factor alpha (TNF alpha) led to a marked reduction of membrane potential. Here we report that this depolarization depends on activation of protein kinase C (PKC), since it could be blocked by the PKC antagonists staurosporine and H7 and it could be mimicked by direct activation of PKC using the phorbol ester phorbol 12-myristate 13 acetate (PMA). Analyses of whole cell currents revealed a reduction of inwardly rectifying K+ currents whereas K+ outward currents were not affected. We conclude that TNF alpha induces changes of basic electrophysiological properties of astrocytes which are similar to those induced by proliferation or an in vitro model of traumatic injury.

Update on the thyroid sodium iodide symporter: a novel thyroid antigen emerging on the horizon.

The ability to transport and concentrate iodide, a fundamental property of normally functioning thyroid epithelial cells, represents a key step in the production of the iodine-containing thyroid hormones. Iodide uptake across the basolateral membrane of thyroid follicular cells is made possible by the Na ˛ /I π symporter (NIS), an active co-transport mechanism that is driven by an inwardly directed Na ˛ gradient. In addition to providing a mechanism for several diagnostically useful tests such as the thyroidal radioiodine uptake test and the perchlorate discharge test, NIS-mediated active I π accumulation by thyroid tissue is a crucial requirement for the success of radioactive iodine therapy. As highlighted earlier (1), cloning of the rat NIS from an FRTL-5 cell-derived cDNA library by Carrasco‘s group has revealed an open reading frame of 1854 nucleotides that encodes a 618 amino acid intrinsic membrane protein with 12 putative membrane-spanning domains. This discovery has set the stage for Smanik et al. (2), who subsequently cloned the human homologue of NIS from a human thyroid cDNA library. An open reading frame (nucleotides 348‐2276) of human NIS (hNIS) encodes a protein of 643 amino acids that shares 84% amino acid identity with recombinant NIS (rNIS) and has a predicted molecular mass of 68·7 kDa. Assignment of the 12 transmembrane domains in hNIS has been based on the proposed transmembrane domains of the rNIS, in which the two protein sequences are highly conserved. Transient transfection of COS-7 cells with the hNIS cDNA clone resulted in perchlorate-sensitive iodide uptake, confirming expression of functionally intact hNIS. Using reverse transcriptase-polymerase chain reaction, the authors detected hNIS expression at variable levels in papillary thyroid carcinoma tissues, but not in any of the thyroid carcinoma cell lines that lack iodide uptake activity, suggesting that loss of iodide-concentrating activity in thyroid carcinomas may be due to a loss of hNIS expression. Shortly after cloning of hNIS, Kosugi et al. (3) succeeded in establishing stable expression of recombinant rNIS in a Chinese hamster ovary (CHO) cell line. Subsequent analysis of its basic biochemical and electrophysiological properties, including time course, Na ˛ -dependency, kinetics and competitive inhibition by perchlorate and thiocynate, revealed characteristics

Electrophysiological properties of lumbar motoneurons in the alpha-chloralose-anesthetized cat during carbachol-induced motor inhibition.

The present study was undertaken 1) to examine the neuronal mechanisms responsible for the inhibition of spinal cord motoneurons that occurs in alpha-chloralose-anesthetized cats following the microinjection of carbachol into the nucleus pontis oralis (NPO), and 2) to determine whether the inhibitory mechanisms are the same as those that are responsible for the postsynaptic inhibition of motoneurons that is present during naturally occurring active sleep. Accordingly, the basic electrophysiological properties of lumbar motoneurons were examined, with the use of intracellular recording techniques, in cats anesthetized with alpha-chloralose and compared with those present during naturally occurring active sleep. The intrapontine administration of carbachol resulted in a sustained reduction in the amplitude of the spinal cord Ia monosynaptic reflex. Discrete large-amplitude inhibitory postsynaptic potentials (IPSPs), which are only present during the state of active sleep in the chronic cat, were also observed in high-gain recordings from lumbar motoneurons after the injection of carbachol. During carbachol-induced motor inhibition, lumbar motoneurons exhibited a statistically significant decrease in input resistance, membrane time constant and a reduction in the amplitude of the action potential's afterhyperpolarization. In addition, there was a statistically significant increase in rheobase and in the delay between the initial-segment (IS) and somadendritic (SD) portions of the action potential (IS-SD delay). There was a significant increase in the mean motoneuron resting membrane potential (i.e., hyperpolarization). The preceding changes in the electrophysiological properties of motoneurons, as well as the development of discrete IPSPs, indicate that lumbar motoneurons are postsynaptically inhibited after the intrapontine administration of carbachol in cats that are anesthetized with alpha-chloralose. These changes in the electrophysiological properties of lumbar motoneurons were found to be comparable with those that take place during the atonia of active (rapid-eye-movement) sleep in chronic cats. The present results support the conclusion that the neural system that is responsible for motor inhibition during naturally occurring active sleep can also be activated in alpha-chloralose-anesthetized cats following the injection of carbachol into the NPO.

Adenosine: an effective and safe antiarrhythmic drug in pediatrics.

Adenosine is an effective, safe drug for the diagnosis and treatment of paroxysmal tachycardias in adult and pediatric patients. A starting dose of 0.05-0.10 mg/kg as a rapid bolus injection is recommended for infants and children. An electrophysiologic effect can be expected within 20 seconds after injection. Dosage may be increased up to 0.3 mg/kg in steps of 0.05-0.10 mg/kg or until conversion to sinus rhythm is reached. Due to its basic electrophysiologic properties of slowing conduction in the atrioventricular (AV) node, which may result in transient AV block, adenosine is almost always effective in terminating supraventricular tachycardias in which the AV node forms a critical part of the reentrant circuit (i.e., AV nodal reentrant tachycardia and AV reciprocating tachycardia). Based on its properties, adenosine is also advocated as a useful diagnostic tool for unmasking primary atrial tachycardias by inducing transient high grade AV block. Advantages over other antiarrhythmic agents include the agent's short half-life (<2 seconds) and minimal or no negative influence on blood pressure. Because of its short half-life, however, early recurrence of the tachycardia is observed in up to one-third of patients treated. Based on rare but serious unwanted side effects, patients with known or suspected irritable airways and sinus node dysfunction and those who have undergone orthotopic cardiac transplantation should probably not be given adenosine. Adenosine may be recommended as the drug of choice for treatment of paroxysmal tachycardia in young patients. Primary success rates range between 85% and 100% of all the tachycardia episodes treated. Termination of the tachydysrhythmia, however, does not always mean that the underlying dysrhythmia was of supraventricular origin with the AV node as a critical part of the tachycardia mechanism. Rare but possible life-threatening side effects (prolonged sinus arrest and complete AV block, atrial fibrillation, acceleration of ventricular tachycardia, apnea) necessitate proper monitoring of the patients.

An in vitro model of the kitten retinogeniculate pathway.

An organotypic explant coculture method is described for the developing retinogeniculate pathway of the cat. Retinal explants and thalamic slices containing the dorsal lateral geniculate nucleus (LGN), derived from early postnatal kittens, can be grown in serum-free culture medium for several days. In such cultures, retinal ganglion cells (RGCs) and LGN neurons retained their age-specific morphological features and developed functional connections. Labeling of RGCs and their processes with DiI showed that all three major classes of RGCs (alpha/Y, beta/X, gamma/W) were present in cocultured retinal explants. Retinal axons readily regenerated into thalamic slices and, over time, developed arbors within the LGN. Retrograde labeling from the LGN traced the origin of these axons almost exclusively to alpha-cells in the retina. In vitro intracellular recordings indicated that LGN cells maintained their basic electrophysiological properties in coculture. Current injection generated action potentials, and, at hyperpolarized levels, it led to low-threshold Ca2+ spiking. Regenerated retinal axons also formed functional connections with LGN neurons. Electrical stimulation of the retinal explant elicited excitatory postsynaptic responses (EPSPs) in LGN cells. Drop application of specific glutamate antagonists indicated that EPSPs had both N-methyl-D-aspartate (NMDA) and non-NMDA receptor components. The morphology of the LGN neurons was examined after intracellular injections of biocytin during recording. Labeled cells were very similar to those of early postnatal kittens. Although, in general, they had relatively small soma and simple dendritic branching patterns, a few could be recognized as X- or Y-cells. Thus the coculture model can be used to assay the regenerative propensity of different types of RGCs during development.

Rhythmogenesis in single cells and population models: olfactory bulb and hippocampus

Dynamics of single cells, small networks and large cell populations are the subject of investigation. The generation and propagation of action potentials in the two major cell types of the olfactory bulb, i.e. in the mitral and granule cells, are simulated by multi-compartmental modeling techniques. The specific effects of the individual ionic currents, the propagation of the signals through the compartments, and dynamic phenomena occurring in small networks (such as synchronized oscillation due to excitatory and inhibitory coupling) have been demonstrated. A statistical model is given to describe the electrical activity patterns of large neural populations. The model is applied for describing the CA3 region of the hippocampus by incorporating some basic electrophysiological properties of hippocampal pyramidal and inhibitory neurons. Population activities as well as underlying single cell voltages are simulated during population bursts in the model of disinhibited hippocampal CA3 slice.

Modulatory Actions of Dopamine on NMDA Receptor-Mediated Responses Are Reduced in D1A-Deficient Mutant Mice

The role of D1 dopamine (DA) receptors in mediating the ability of DA to modulate responses attributable to activation of NMDA receptors was examined in mice lacking D1A dopamine receptors. Specifically, experiments were designed to test the hypothesis that the ability of DA to potentiate responses mediated by activation of NMDA receptors was attributable to activation of D1 receptors. Based on this hypothesis, we would predict that in the D1A mutant mouse, either DA would not induce enhancement of NMDA-mediated responses, or the enhancement would be severely attenuated. The results provided evidence to support the hypothesis. In mutant mice, DA and D1 receptor agonists did not potentiate responses mediated by activation of NMDA receptors. In contrast, in control mice, both DA and D1 receptor agonists markedly potentiated responses mediated by activation of NMDA receptors. The effects of DA in attenuating responses mediated by activation of non-NMDA receptors also were altered in the mutant, suggesting that this action of DA may require coupling or interactions between D1 and D2 receptors. The present studies also provided an opportunity to assess some of the basic electrophysiological and morphological properties of neostriatal neurons in mice lacking D1A DA receptors. Resting membrane potential, action potential parameters, input resistance, excitability, somatic size, dendritic extent, and estimates of spine density in mutants and controls were similar, suggesting that these basic neurophysiological and structural properties have not been changed by the loss of the D1A DA receptor.

Tumour necrosis factor-alpha increases intracellular Ca2+ and induces a depolarization in cultured astroglial cells.

Tumour necrosis factor (TNF)-alpha, a strong immune mediator, is released within the brain during inflammatory diseases and contributes to immunological activation of glial cells. Here we report that, in astrocytes, TNF-alpha also affects the intracellular Ca2+ homeostasis and basic electrophysiological properties such as the membrane potential. Using the Ca2+ indicator dye fura-2 in a cell culture model, we found that TNF-alpha (10-1000 U ml-1), but not interleukin 1 or 6, induced a slow but more than two-fold increase of the intracellular Ca2+ concentration, which could be blocked by Co2+ (1.0 mM), verapamil (100 microM) or omission of external Ca2+. This intracellular Ca2+ increase was accompanied by a marked decrease of the membrane potential by 35 mV. CSF of patients with bacterial meningitis, known to contain large amounts of TNF-alpha, induced a similar depolarization of astrocytes, which was markedly reduced by a neutralizing anti-TNF-alpha antibody. We conclude that TNF-alpha induces an increase of intracellular Ca2+ and a depolarization in astrocytes with the consequence of disturbing voltage-dependent glial functions such as regulation of local ion concentrations and glutamate uptake. During inflammatory CNS diseases this immuno-electrical coupling may contribute to an impairment of neuronal function.

Changes in the electrophysiological properties of cat spinal motoneurons following the intramuscular injection of adriamycin compared with changes in the properties of motoneurons in aged cats

1. This study was undertaken to investigate the effects of adriamycin (ADM, Doxorubicin) on the basic electrophysiological properties of spinal cord motoneurons in the adult cat. ADM was injected into the biceps, gastrocnemius, semitendinosus, and semimembranosus muscles of the left hindlimb (1.2 mg per muscle). Intracellular recordings from motoneurons innervating these muscles were carried out 12, 20, or 40 days after ADM administration and from corresponding motoneurons in untreated control cats. 2. Twelve days after ADM injection, motoneurons innervating ADM-treated muscles (ADM MNs) exhibited statistically significant increases in input resistance, membrane time constant, and amplitude of the action potential's afterhyperpolarization (AHP). In addition, there was a statistically significant decrease in rheobase and in the delay between the action potential of the initial segment (IS) and that of the somadendritic (SD) portion of the motoneuron (IS-SD delay). There were no significant changes in the resting membrane potential, threshold depolarization, action potential amplitude, or axonal conduction velocity. 3. The changes in electrical properties of motoneurons at 20 and 40 days after ADM injection were qualitatively similar to those observed at 12 days. However, at 40 days after ADM injection there was a statistically significant decrease in the axonal conduction velocity of the ADM MNs. 4. The normal correlations that are present between the AHP duration and electrical properties of the control motoneurons were observed in the ADM MNs, e.g., AHP duration was positively correlated with the input resistance and time constant and negatively correlated with the axonal conduction velocity. The correlation coefficients, however, were reduced in comparison with the control data. 5. This study demonstrates that ADM exerts significant effects on the electrical properties of motoneurons when injected into their target muscles. The majority of the changes in motoneuron electrical properties caused by ADM resemble those observed in motoneurons of aged cats. Additional research is required to determine whether the specific changes induced in motoneurons by ADM and those that occur in motoneurons in old age are due to similar degradative mechanisms.

Slowly inactivating outward currents in a cuticular mechanoreceptor neuron of the cockroach (Periplaneta americana).

1. Although rapid adaptation is a widespread feature of sensory receptors, its ionic basis has not been clearly established in any touch receptor, because their small sizes have severely restricted the range of experiments tat can be performed. In the cockroach tactile spine, intracellular voltage-clamp recordings are now possible. 2. The basic electrophysiological properties of the cockroach femoral tactile spine neuron were studied using discontinuous (switching) single-electrode current- and voltage-clamp recordings. A slowly inactivating voltage-sensitive K+ outward current was detected after the major inward currents were blocked with tetrodotoxin. 3. The total outward current activated in < 1 ms at voltages above 0 mV. At moderate depolarizations it did not inactivate, but at higher depolarizations an inactivation time constant of approximately 260 ms was measured. Some recordings also revealed an additional, slower inactivation time constant of approximately 2.5 s. 4. More than half of the voltage-sensitive K+ outward current could be blocked with the Ca2+ channel blockers Co2+ and Cd2+. Tetraethylammonium chloride (TEA) also reduced the amplitude of the outward current to about half of its original amplitude. The actions of both blockers were reversible and probably reflect overlapping blockades of two separate outward currents. 5. The reversal potentials of the currents that remained after block with Co2+ (-91.7 mV) or TEA (-86.8 mV) were both near the K+ equilibrium potential expected for the tactile spine neuron. The voltage dependencies of activation of the Co(2+)- and TEA-resistant currents were both well fitted by Boltzmann distributions, giving values of half maximal activation (V50) equal to -34.5 mV for the Co(2+)-resistant current and -51.3 mV for the TEA-resistant current. 6. Current-clamp recordings revealed that the TEA-sensitive K+ current was the major component of action potential repolarization but that it did not effect the frequency of action potentials evoked by steady depolarization. On the other hand, blockers of Ca(2+)-sensitive K+ currents (Cd2+, Co2+, or charybdotoxin) reduced adaptation and increased the frequency of action potentials significantly but did not effect the duration or amplitude of individual action potentials.

Several types of Ca2+ channels mediate glutamatergic synaptic responses to activation of single Thy-1-immunolabeled rat retinal ganglion neurons

A dissociated cell culture from the postnatal rat retina was established to characterize the synapses formed by retinal ganglion neurons (RGNs) in vitro. An antibody against Thy-1.1 was used to preselect putative RGNs for pair patch-clamp recording with the principal aim of identifying the released transmitter(s) and estimating the role of different types of voltage-activated Ca2+ channels in evoked transmitter release. The population of Thy-1+ neurons was heterogeneous. Staining patterns, soma-dendritic geometries and axon length displayed variations that could be related to basic electrophysiological properties, such as amplitudes of voltage-activated Na+ currents (INa(V)), action potential size and capacity for repetitive discharge. Out of 73 coupled connections, 33 pairs were glutamatergic. With no exception, these connections were formed by the axons of strongly labeled Thy-1+ neurons with large INa(V) (typically > 2 nA) and repetitive firing over a broad current range. Such neurons were classified as RGNs. Forty out of 73 coupled pairs were GABAergic. These connections were always formed by weakly stained Thy-1+ neurons with small INa(V) (typically < 2 nA) and very limited capacity for repetitive discharge. Such neurons were tentatively classified as displaced amacrine cells. Evoked EPSCs in response to RGN activation were completely blocked by low concentrations of Cd2+ or Gd3+. omega-CgTx-GVIA (5 microM) reduced EPSCs to 67 +/- 29%, omega-AgaTx-IVA (200 nM) had no effect, and nifedipine (15 microM) enhanced the evoked EPSCs. Our experiments indicate that (1) the transmitter released by RGNs is glutamate and (2) the major part of synaptic glutamate release is governed by a novel toxin-resistant Ca2+ channel. The results further suggest that the characteristic phenotype of RGNs is well maintained in dissociated cell culture. In conjunction with electrophysiological tests Thy-1+ labeling can be used for RGN identification.

Computer aided development of antiarrhythmic agents with class IIIa properties.

Most antiarrhythmic agents were discovered accidentally. In the last decade, the understanding of the mechanisms of action of agents with electrophysiologic activity has progressed greatly. As a result, it was possible to compute, before the CAST trial, that the agents selected for the trial would not be effective against tachycardias and that the drugs would be unsafe. Extension of these computations to existing Class I agents indicated that they were all poor suppressors of ventricular tachycardia. Furthermore, a Class I agent with an optimal electrophysiologic profile still computes to be a two-edged sword, possessing both antiarrhythmic and proarrhythmic properties. Fortunately, it is possible to conceive of drug profiles that would be purer antiarrhythmic agents. For example, a drug that only upon the development of a tachycardia lengthens action potential duration in a use-dependent manner until the refractory period exceeds the tachycardia cycle length will render continuation of the tachycardia impossible. Recognition of chemicals that have Class IIIa properties with the appropriate kinetics is a challenging task. However, today's microprocessors have become powerful enough to characterize the Class III kinetics. A system that fully automatically screens for effective antiarrhythmic agents is described. It is expected that chemicals selected for optimal basic electrophysiologic properties will yield safer and more effective antiarrhythmic agents.

The effect of quinidine and myocardial ischemia on the isolated rat heart with fat-free diet.

Fat-free diet changes the lipid content and the electrophysiological properties of the rat myocardium. Five percent fat supplementation to the diet does not alter the basic electrophysiological properties but still has a biochemical effect on the lipid content of the myocardium. The purpose of this work was to determine whether these biochemical alterations affect the response of the myocardium to quinidine and ischemia, both of which interact with the lipid component of the membrane. We used strength-duration, strength-interval and threshold of ventricular fibrillation to measure the electrophysiological properties of the isolated rat heart at baseline and after 30 minutes of quinidine perfusion or coronary artery ligation. The fatty acid composition of the myocardium was analyzed. We found that a fat-free diet caused essential fatty-acid deficiency, while 5% fat supplementation had a partial protective effect. Quinidine decreased excitability and increased refractoriness in both groups but had more effect on the fat-free diet hearts group. There was no difference in the ventricular fibrillation threshold. Ischemia increased myocardial excitability in the fat-free diet hearts group and had no effect on refractoriness or ventricular fibrillation threshold. These results support the theory that the lipid composition of the myocardial membrane affects its response to lipophilic drugs and ischemia.

Actions of cisplatin on the electrophysiological properties of cultured dorsal root ganglion neurones from neonatal rats.

In this study we have employed the whole cell patch clamp technique to investigate the effects of an anti-cancer drug cisplatin on basic electrophysiological properties of cultured dorsal root ganglion neurones from neonatal rats. The results show that within the clinical concentration range, cisplatin (0.1 to 10 microM) caused a decrease in input conductance, and complex changes in resting membrane potential in these cultured sensory neurones. The dominant effects of cisplatin on input conductance may be due to inhibition of leak conductances. Transplatin (5 microM) was significantly less effective than cisplatin at reducing input conductance which suggests a degree of stereoselectivity. Cisplatin (1 to 5 microM) transiently increased excitability of the cultured neurones as reflected by a reduction in the threshold for activation of action potentials by 8 mV. The rise time, peak amplitude and duration of action potentials were not changed by acute application of 5 microM cisplatin. Long term treatment of neurones with cisplatin (5 microM), for up to 1 week reduced the viability of the cultures, and attenuated neurone excitability, although input conductance of the cells was significantly increased to 322 +/- 49 M omega (n = 9) compared with controls of 210 +/- 20 M omega (n = 30; P < 0.05). Acute and chronic treatment of cultured neurones with cisplatin therefore produced contrasting actions.

Characteristics of transient outward current in human ventricular myocytes from patients with terminal heart failure.

A variety of outward currents exists in ventricular myocardium of different species influencing action potential duration and electrical activity. Transient outward currents have been reported in ventricular tissue of some animals but are small or absent in others. This study was conducted to investigate whether a transient outward current exists in human ventricular myocardium and to characterize its basic electrophysiological properties. Currents were recorded from enzymatically isolated human ventricular myocytes obtained from explanted hearts of 22 patients with terminal heart failure. In almost all cells studied, a transient outward current could be recorded on depolarization to between -20 and +80 mV. The size of the transient outward current was usually large enough to mask the Ca2+ current. It could be recorded under conditions in which Ca2+ influx and intracellular Ca2+ transients were suppressed. Basic current characteristics were similar to transient outward currents observed in other species. Inactivation of the transient outward current was monoexponential, with a time constant of 54.8 +/- 3.7 milliseconds at +40 mV. Half-maximal activation occurred at 16.7 +/- 1.6 mV; half-maximal steady-state inactivation occurred at -34.5 +/- 2.3 mV. Frequency-dependent reduction of peak transient outward current was 29.8 +/- 1.4% at 2 Hz compared with resting conditions. Recovery from inactivation was voltage dependent and had a biexponential time course; the faster time constant (41.0 +/- 6.5 milliseconds at -80 mV) accounted for 86.0 +/- 5.2% of total current. The transient outward current was sensitive to 4-aminopyridine (IC50, 1.15 mM). These results indicate that a large Ca(2+)-independent transient outward K+ current is present in human ventricular myocytes that might be regulated by physiological or pathological events and is a potential site for pharmacological intervention.