Chris P. Bolter

Influence of right atrial pressure on the cardiac pacemaker response to vagal stimulation

We have recently shown that the intrinsic rate response to an increase in right atrial pressure is augmented when cardiac muscarinic receptors are activated. This present study examines the cardiac pacemaker response to vagal stimulation at different values of right atrial pressure in isolated rat right atrium and in the rabbit heart in situ. In the rat atrium, when pressure was raised in steps from 2 to 10 mmHg, there was a progressive reduction in the response to vagal stimulation [40.5 +/- 7.2% reduction (mean +/- SE) at 8 mmHg, P < 0.01], which was independent of the level of vagal bradycardia, that persisted in the presence of the beta-adrenergic agonist isoproterenol. In barbiturate-anesthetized rabbits with cervical vagi cut and beta-adrenergic blockade, raising right atrial pressure approximately 2.5 mmHg by blood volume expansion reduced the bradycardia elicited by electrical stimulation of the peripheral end of the right vagus nerve (9.1 +/- 1.1% reduction, P < 0.0001). These results demonstrate that vagal bradycardia is modulated by the level of right atrial pressure and suggest that normally right atrial pressure may interact with cardiac vagal activity in the control of heart rate.

Do cardiac neurons play a role in the intrinsic control of heart rate in the rat

Three experiments were performed to see whether cardiac neurons contribute to the intrinsic control of heart rate in right atria of adult rats. The intrinsic heart rate response (IRR) was examined by raising right atrial pressure from 2 to 8 mmHg for 3 min. In isolated preparations of the right atrium, the IRR was not significantly altered by the addition of either 1 μM atropine (n = 6; control +19 ± 3 min−1; atropine +18 ± 3 min−1 (mean ± S.E.M.)) or 1 μM propranolol (n = 5; control +22 ± 4 min−1; propranolol +21 ± 3 min−1). Tetrodotoxin (0.5 μM) had no effect on the IRR (n = 6; control +37 ± 5 min−1; tetrodotoxin 38 ± 5 min−1). In another experiment, 2‐day‐old rat pups were injected with capsaicin (50 mg kg−1; treated) or with vehicle (control). There was no difference in the IRR of right atrial preparations taken from control and treated animals after they reached adulthood (control (n = 7) and treated (n = 8): +30 ± 4 and +32 ± 4 min−1). The influence of right atrial pressure on the efficacy of vagal stimulation was examined. The rate response to vagal stimulation was reduced similarly in control and treated preparations when pressure was elevated from 2 to 4 mmHg (control and treated: ‐34 ± 5% and ‐33 ± 3%). The effectiveness of the capsaicin treatment was confirmed by the depletion of substance P‐immunoreactive nerve fibres in cardiac tissues. Together, these results strongly suggest that cardiac neurons are not involved in intrinsic heart rate control.

The influence of heart rate on baroreceptor fibre activity in the carotid sinus and aortic depressor nerves of the rabbit

The arterial baroreceptors and their afferent fibres provide the sensory arm of the reflex that regulates systemic arterial pressure. We have examined whether the relationship between mean baroreceptor discharge and mean arterial pressure is altered when heart rate changes. Experiments were performed on pentobarbitone‐anaesthetized rabbits. We recorded the activity of single and multifibre preparations of the carotid sinus (CSN) and aortic depressor nerves (ADN). Data were collected under control conditions and while heart rate was increased by ∼30–35% by right atrial pacing. Baroreceptor regions were exposed to ramps of pressure (from ∼25 to 140 mmHg, at ∼0.5–1 mmHg s−1), generated by inflation and deflation of cuffs placed around the inferior vena cava and descending thoracic aorta. Response curves relating baroreceptor discharge to mean pressure were constructed and fitted with third‐order polynomial expressions. To provide a measure of an effect of an increase in heart rate on the response curve in the region of the normal operating pressure, we calculated the position of the test response curve relative to the position of the control curve at 90 mmHg (δBP90). For the ADN, the activity of single fibres (presumptive myelinated fibres) was unaffected by increasing heart rate (δBP90=+0.1 ± 1.0 mmHg), while single fibres in the CSN showed a small increase in activity (δBP90=−1.5 ± 0.3 mmHg). In multifibre preparations there was a small increase in activity that may be attributable to additional activity in unmyelinated fibres (ADN, δBP90=−3.4 ± 1.2 mmHg; CSN, δBP90=−5.2 ± 0.9 mmHg). We conclude that the mean discharge of arterial baroreceptors remains a reliable index of mean arterial pressure in the presence of substantial changes in heart rate.

The effects of tertiapin-Q on responses of the sinoatrial pacemaker of the guinea-pig heart to vagal nerve stimulation and muscarinic agonists.

Using Langendorff preparations of the guinea‐pig heart, we have examined the participation of the acetylcholine (ACh)‐activated potassium channel, IK,ACh, in the bradycardia produced by electrical stimulation of the vagus (parasympathetic) nerve and muscarinic agonists (ACh and bethanecol, bolus i.a.). Hearts from young animals (160–250 g) were perfused with Krebs–Henseleit solution, and pacemaker frequency was determined from the P wave of an ECG. Tertiapin‐Q was used to block IK,ACh. Vagal stimulation (10 s trains at 2, 5 and 10 Hz) produced graded reductions in atrial rate that were substantially attenuated, and to a similar extent, by 300 nm and 1 μm tertiapin‐Q (to 0.42 ± 0.12, mean ±s.d., of the control values; P < 0.001). Acetylcholine (3 nmol) produced brief graded bradycardias that were also attenuated by tertiapin‐Q (0.24 ± 0.24; P= 0.006). Similar results were obtained when experiments were repeated in 2 mm Cs+ (to block the hyperpolarization‐activated pacemaker current). Bethanecol (30, 50 and 70 nmol), a muscarinic agonist with no appreciable nicotinic activity, produced sustained bradycardias that were attenuated by 300 nm tertiapin‐Q (0.36 ± 0.21; P < 0.0001). The responses to vagal stimulation and ACh developed more slowly in tertiapin‐Q, indicating that a rapidly acting mechanism had been blocked. Responses to vagal stimulation were faster in 2 mm Cs+. Together, these observations show that ACh released from parasympathetic nerve varicosities exerts a considerable part of its effect on the pacemaker by activating IK,ACh and acts in a manner not readily distinguishable from that of directly applied muscarinic agonists.

The intrinsic rate response of the isolated right atrium of the rat, Rattus norvegicus.

Experiments were performed on rat right atria maintained at 37 degrees C in oxygenated Krebs-Henseleit solution, at a baseline diastolic transmural pressure of 2 mmHg. A step increment in right atrial pressure caused an increase in atrial rate which reached a steady value after 2-3 min (rate response). An 8-mmHg increase in atrial pressure caused an 8% increase in atrial rate (n = 9, P < 0.01). When the atrial rate was reduced by carbamylcholine, the rate response was augmented. After a 34% reduction in atrial rate, an 8-mmHg increase in atrial pressure increased atrial rate by 51% (n = 7, P < 0.01). When atrial rate was elevated 71% by isoprenaline, the rate response was reversed (atrial rate decreased 3% following an 8-mmHg increase in atrial pressure; n = 7, P < 0.01). In another series of experiments, atrial rate was adjusted to a wide range of values by exposure to carbamylcholine and isoprenaline, first applied singly, and then in combination. At any given atrial rate, the rate response was always larger when both agonists were present; this difference was greatest when atrial rate was near the control untreated value.

The muscarinic-activated potassium channel always participates in vagal slowing of the guinea-pig sinoatrial pacemaker

UNLABELLED Controversy persists regarding participation of the muscarinic-activated potassium current (c(KACh)) in small and moderate vagal bradycardia. We investigated this by (i) critical examination of earlier experimental data for mechanisms proposed to operate in modest vagal bradycardia (modulation of I(f) and inhibition of a junctional Na(+) current) and (ii) experiments performed on isolated vagally-innervated guinea-pig atria. In 8 superperfused preparations, 10-s trains of vagal stimulation (1 to 20Hz) produced a bradycardia that ranged from 1 to 80%. Hyperpolarisation of sinoatrial cells accompanied bradycardia in 65/67 observations (linear correlation between bradycardia and increase in maximum diastolic potential (mV)=0.076x%; R(2)=0.57; P<0.001). In bath-mounted preparations single supramaximal stimuli to the vagus immediately and briefly increased pacemaker cycle length in 7 of 18 preparations. This response was eliminated by 300nM tertiapin-Q. Trains of 10 single supramaximal vagal stimuli applied at 1-s intervals caused progressive increase in overall cycle length during the train; immediate and brief increases in cycle length occurred following some stimuli. Immediate brief responses and part of the slower response to the stimulus train were removed by 300nM tertiapin-Q. SUMMARY experimental data shows that small and modest vagal bradycardia is accompanied by hyperpolarisation of the pacemaker cell which is severely attenuated by tertiapin-Q. These observations support the idea that activation of I(KACh) occurs at all levels of vagal bradycardia. Contradictory conclusions from earlier studies may be attributed to the nature of experimental models and experimental design.

Tertiapin-Q removes a mechanosensitive component of muscarinic control of the sinoatrial pacemaker in the rat.

1. In an isolated right atrial preparation, an increase in right atrial pressure (RAP) produces an increase in atrial rate. This rate response is larger and occurs faster when there is background vagal or muscarinic stimulation.

Failure of Ba2+ and Cs+ to block the effects of vagal nerve stimulation in sinoatrial node cells of the guinea-pig heart

The aim of the study was to evaluate which ionic currents are modified in the sinoatrial node of guinea pigs when the vagus is stimulated. Responses of isolated atrial preparations to bilateral vagus nerve stimulation were examined. In bath-mounted preparations, 10-s trains of vagal stimulation (1-50 Hz) slowed the rate at which atrial contractions occurred. After the trains of stimuli, the force generated by each contraction was reduced. Both vago-inhibitory responses persisted in the presence of caesium (2 mM) and barium ions (1 mM). Vagal stimulation evoked a similar bradycardia in superperfused preparations in which intracellular recordings were made from pacemaker cells in the sinoatrial node. When pacemaking activity was abolished by adding the organic calcium channel antagonist nifedipine (1 microM) to the perfusate, vagal stimulation generated an inhibitory junction potential (IJP). Both the bradycardia and the amplitude of the inhibitory junction potential increased as the frequency of vagal stimulation was increased. The ability of vagal stimulation to produce inhibitory junction potentials was unaffected by the addition of caesium and barium ions to the perfusate. These observations suggest that the negative chronotropic and inotropic responses to vagal stimulation only minimally involve a muscarinically activated potassium current (I(KACh)) or changes in the hyperpolarization-activated pacemaker current Ih.

Tertiapin-Q removes a large and rapidly acting component of vagal slowing of the guinea-pig cardiac pacemaker

The participation of acetylcholine-activated potassium current (I(K,ACh)) and hyperpolarization-activated pacemaker current (I(f)) in vagal bradycardia were examined using vagally-innervated preparations of guinea-pig atria. Preparations were maintained in Krebs-Henseleit solution (36 degrees C). Before treatment, trains of vagal stimuli (10 s at 2, 5 and 10 Hz) produced graded bradycardias displaying rapid onset and offset. Tertiapin-Q (300 nM), which blocks I(K,ACh), had no effect on baseline atrial rate. In tertiapin-Q, vagal bradycardia displayed a gradual onset and offset, with a peak response ~50% of that recorded in control conditions. Cumulative addition of 1 mM ZD7288 (blocker of I(f)) caused atrial rate to fall by ~60%, but had no further effect on the amplitude of the vagal bradycardia, while response onset and offset became slightly faster. From these observations, we argue that (i) vagal bradycardia was attributable primarily to activation of I(K,ACh), (ii) vagal modulation of I(f) had a minor influence on the rate of onset and offset of bradycardia, and (iii) removal of the influence of I(K,ACh) unmasked a slow response, of undetermined origin, to vagal stimulation. In a separate set of experiments we compared the effects of 1 mM Ba(2+) and 300 nM tertiapin-Q on vagal bradycardia. Ba(2+) reduced baseline atrial rate and the response to vagal stimulation. Subsequent cumulative addition of tertiapin-Q had no additional effect on baseline atrial rate, but caused further reduction in the amplitude of vagal bradycardia, suggesting that 1 mM Ba(2+) did not achieve a complete block of I(K,ACh) in this preparation.

Interaction of the autonomic nervous system with intrinsic cardiac rate regulation in the guinea-pig, Cavia porcellus.

In the denervated mammalian heart a change in right atrial pressure will still alter heart rate (intrinsic rate response, IRR). We have examined the IRR in isolated right atria of the guinea-pig maintained in oxygenated Krebs-Henseleit solution at 37 degrees C, to compare with and extend studies in other species, and to determine whether the guinea-pig is a suitable model for electrophysiological studies of the IRR. Baseline diastolic transmural pressure was set at 2 mmHg. A 6-mmHg increase in right atrial pressure (RAP) caused an increase in atrial rate that reached a steady value of 15 min(-1) after 1-2 min. This response was enhanced by carbamylcholine and attenuated by isoprenaline. The influence of RAP on the rate response to vagal stimulation was examined. With RAP set at 8 mmHg, the reduction in atrial rate following vagal stimulation was 72+/-5% of that at 2 mmHg (n=6, mean+/-S.E., P<0.005). Continuous vagal stimulation produced a sustained bradycardia, and the effect of this bradycardia on the IRR was examined. When atrial rate was reduced 6% by vagal stimulation, the IRR was augmented to 202+/-21% of the control (n=6, P<0.005). This augmentation was larger (P<0.05) than that seen when atrial rate was reduced 8% by carbamylcholine (130+/-8% of control; n=7, P<0.05). Overall, the IRR in the guinea-pig is similar to that in the rabbit, and shows similar interactions with the autonomic nervous system.