Effects of Acetylcholine and Noradrenalin on Action Potentials of Isolated Rabbit Sinoatrial and Atrial Myocytes

The autonomic nervous system controls heart rate and contractility through sympathetic and parasympathetic inputs to the cardiac tissue, with acetylcholine (ACh) and noradrenalin (NA) as the chemical transmitters. In recent years, it has become clear that specific Regulators of G protein Signaling proteins (RGS proteins) suppress muscarinic sensitivity and parasympathetic tone, identifying RGS proteins as intriguing potential therapeutic targets. In the present study, we have identified the effects of 1 μM ACh and 1 μM NA on the intrinsic action potentials of sinoatrial (SA) nodal and atrial myocytes. Single cells were enzymatically isolated from the SA node or from the left atrium of rabbit hearts. Action potentials were recorded using the amphotericin-perforated patch-clamp technique in the absence and presence of ACh, NA, or a combination of both. In SA nodal myocytes, ACh increased cycle length and decreased diastolic depolarization rate, whereas NA decreased cycle length and increased diastolic depolarization rate. Both ACh and NA increased maximum upstroke velocity. Furthermore, ACh hyperpolarized the maximum diastolic potential. In atrial myocytes stimulated at 2 Hz, both ACh and NA hyperpolarized the maximum diastolic potential, increased the action potential amplitude, and increased the maximum upstroke velocity. Action potential duration at 50 and 90% repolarization was decreased by ACh, but increased by NA. The effects of both ACh and NA on action potential duration showed a dose dependence in the range of 1–1000 nM, while a clear-cut frequency dependence in the range of 1–4 Hz was absent. Intermediate results were obtained in the combined presence of ACh and NA in both SA nodal and atrial myocytes. Our data uncover the extent to which SA nodal and atrial action potentials are intrinsically dependent on ACh, NA, or a combination of both and may thus guide further experiments with RGS proteins

Mechanisms of inherited cardiac conduction disease

Cardiac conduction disease (CCD) is a serious disorder of the heart. The pathophysiological mechanisms underlying CCD are diverse. In the last decade the genes responsible for several inherited cardiac diseases associated with CCD have been identified. If CCD is of an inherited nature (ICCD), its underlying mechanism can be either structural, functional or there can be overlap between these two mechanisms. If ICCD is structural in nature, it is often secondary to anatomical or histological abnormalities of the heart. Functional ICCD is frequently found as a "primary electrical disease" of the heart, i.e. resulting from functionally abnormal, or absent proteins encoded by mutated genes, often cardiac ion channel proteins involved in impulse formation. It can thus be hypothesised that patients with inherited structural or functional ICCD suffer from fundamentally different diseases. It is worthwhile to consider this hypothesis, since it could have implications for diagnosis, treatment, prognosis and, possibly, for the patient's relatives. In this review we aim to find evidence for the idea that functional and structural ICCD are fundamentally different diseases and, if so, whether this has diagnostic and clinical consequences. (c) 2005 Published by Elsevier Ltd on behalf of The European Society of Cardiolog

Action potential transfer at the Purkinje - Ventricular junction: Role of transitional cells

At the Purkinje (P) - ventricular (V) junction a zone of "transitional (T)" cells is found. In the present study we investigated the role of these T cells in P-to-V conduction. Using the "model clamp" technique, an experimentally recorded rabbit P cell was coupled to a phase-2 Luo and Rudy (LR) model cell, which in turn was coupled to a strand of phase-2 LR model cells. In our experiments, the single LR model represents the T cell, while the strand of LR models represents subendocardial V cells. This approach enabled us to change selectively coupling conductance (Gc) between cells, presence of T cell, and relative size of cells. We demonstrated that: 1) a decrease of Gc between P-T and T-V increases the delay of V activation, 2) the delay of V activation is importantly due to conduction between T and V cells, 3) there is a critical Gc for successful conduction at the P-V junction, 4) the critical value of Gc for conduction at the P-V junction is lower in presence (11.0±0.7 nS) than in absence (13.7±0.8 nS) of the T cell, and 5) enlargement of the T zone size hampers successful P-to-V conduction

Contribution of sodium channel mutations to bradycardia and sinus node dysfunction in LQT3 families

One variant of the long-QT syndrome (LQT3) is caused by mutations in the human cardiac sodium channel gene. In addition to the characteristic QT prolongation, LQT3 carriers regularly present with bradycardia and sinus pauses. Therefore, we studied the effect of the 1795insD Na+ channel mutation on sinoatrial (SA) pacemaking. The 1795insD channel was previously characterized by the presence of a persistent inward current (I-pst) at -20 mV and a negative shift in voltage dependence of inactivation. In the present study, we first additionally characterized I-pst over the complete voltage range of the SA node action potential (AP) by measuring whole-cell Na+ currents (I-Na) in HEK-293 cells expressing either wild-type or 1795insD channels. I-pst for 1795insD channels varied between 0.8+/-0.2% and 1.9+/-0.8% of peak I-Na. Activity of 1795insD channels during SA node pacemaking was confirmed by AP clamp experiments. Next, I-pst and the negative shift were implemented into SA node AP models. The -10-mV shift decreased sinus rate by decreasing diastolic depolarization rate, whereas I-pst decreased sinus rate by AP prolongation, despite a concomitant increase in diastolic depolarization rate. In combination, moderate I-pst (1% to 2%) and the shift reduced sinus rate by approximate to10%. An additional increase in I-pst could result in plateau oscillations and failure to repolarize completely. Thus, Na+ channel mutations displaying an I-pst or a negative shift in inactivation may account for the bradycardia seen in LQT3 patients, whereas SA node pauses or arrest may result from failure of SA node cells to repolarize under conditions of extra net inward curren