Bilateral Stellate Ganglionectomy Research Articles

Functional characterization and identification of a therapeutic for a novel SCN5A-F1760C variant causing type 3 long QT syndrome refractory to all guideline-directed therapies

Pathogenic variants in the SCN5A-encoded Nav1.5 sodium channel cause type 3 long QT syndrome (LQT3). We present the case of an infant with severe LQT3 who was refractory to multiple pharmacologic therapies as well as bilateral stellate ganglionectomy. The patient's novel variant, p.F1760C-SCN5A, involves a critical residue of the Nav1.5's local anesthetic binding domain. The purpose of this study was to characterize functionally the p.F1760C-SCN5A variant using TSA-201 and patient-specific induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). Whole-cell patch clamp was used to assess p.F1760C-SCN5A associated sodium currents with/without lidocaine (Lido), flecainide, and phenytoin (PHT) in TSA-201 cells. p.F1760C-SCN5A and CRISPR-Cas9 variant-corrected isogenic control (IC) iPSC-CMs were generated. FluoVolt voltage dye was used to measure the action potential duration (APD) with/without mexiletine or PHT. V1/2 of inactivation was right-shifted significantly in F1760C cells (-72.2 ± 0.7 mV) compared to wild-type (WT) cells (-86.3 ± 0.9 mV; P <.0001) resulting in a marked increase in windowcurrent. F1760C increased sodium late current 2-fold from 0.18% ± 0.04% of peak in WT to 0.49% ± 0.07% of peak in F1760C (P = .0005). Baseline APD to 90% repolarization (APD90) was increased markedly in F1760C iPSC-CMs (601 ± 4 ms) compared to IC iPSC-CMs (423 ± 15 ms; P <.0001). However, 4-hour treatment with 10 μM mexiletine failed to shorten APD90, and treatment with 5μM PHT significantly decreased APD90 of F1760C iPSC-CMs (453 ± 6 ms; P <.0001). PHT rescued electrophysiological phenotype and APD of a novel p.F1760C-SCN5A variant. The antiepileptic drug PHT may be an effective alternative therapeutic for the treatment of LQT3, especially for variants that disrupt the Lido/mexiletine binding site.

Abstract 816: Surgical Bilateral Stellate Ganglionectomy Reduces Mitochondrial Reactive Oxygen Species (mROS) and Prevents Sudden Cardiac Death (SCD) in Non-ischemic Heart Failure (HF)

Introduction: Current therapies for SCD are limited. Bilateral sympathetic stellate ganglionectomy ( BSG ) is a promising new adjunctive therapy in patients with ischemic HF who have incessant ventricular tachyarrhythmias ( VT/VF ) despite optimal medical management. The underlying mechanisms are unknown. The effect of BSG in non-ischemic HF is unclear. Hypothesis: BSG prevents VT/VF and SCD in non-ischemic HF by restoring sympathovagal balance and reducing mROS. Methods: We used a unique pressure-overload guinea pig model that closely mimics human non-ischemic HF, including a high incidence of spontaneous VT/VF/SCD. We randomized the animals to non-failing controls, HF+Sham, or HF+BSG surgery. We used intention to treat analysis to evaluate VT/VF-free survival. We analyzed continuous ECG (VT/VF burden, heart rate variability, QT variability) and echo (cardiac function). In isolated left ventricular (LV) myocytes, we measured Ca 2+ transients and mitochondrial reactive oxygen species (mROS) using targeted ratiometric probes. Results: Half of the HF+Sham animals experienced SCD in 4 weeks. In contrast, BSG abolished SCD, reduced VT/VF, and increased parasympathetic tone (beyond corresponding reductions in sympathetic tone). Fractional shortening of the LV was markedly reduced in HF+Sham (0.28±0.02) vs. control (0.50±0.01), but preserved by BSG (0.45±0.02). Whereas HF+Sham increased mROS levels in LV myocytes (fractional oxidation, F ox : 0.53±0.015) compared to control (0.29±0.020), BSG reduced mROS (0.38±0.035). Mitochondrial antioxidant capacity, as indexed by H 2 O 2 challenge of LV myocytes, was compromised in HF+Sham (F ox : 0.81±0.040) compared to control (0.44±0.061), but preserved by BSG (0.48±0.023). Conclusions: Surgical BSG prevents SCD in non-ischemic HF by reducing mROS, decreasing sympathetic tone, and by increasing parasympathetic tone (possibly by facilitating cholinergic transdifferentiation). This promising new minimally-invasive surgical therapy may be particularly useful in patients with non-ischemic HF on optimal medical therapy who require recurrent ICD shocks to prevent SCD. Patients in the early stages of HF who are not eligible for ICDs but remain at higher SCD risk may also benefit from BSG surgery.

Abstract 115: Surgical Bilateral Stellate Ganglionectomy Reduces Mitochondrial Reactive Oxygen Species (mROS) and Prevents Sudden Cardiac Death (SCD) in Non-ischemic Heart Failure (HF)

Introduction: Current therapies for SCD are limited. Bilateral sympathetic stellate ganglionectomy ( BSG ) is a promising new adjunctive therapy in patients with ischemic HF who have incessant ventricular tachyarrhythmias ( VT/VF ) despite optimal medical management. The underlying mechanisms are unknown. The effect of BSG in non-ischemic HF is unclear. Hypothesis: BSG prevents VT/VF and SCD in non-ischemic HF by restoring sympathovagal balance and reducing mROS . METHODS: We used a unique pressure-overload guinea pig model that closely mimics human non-ischemic HF, including a high incidence of spontaneous VT/VF/SCD. We randomized the animals to non-failing controls, HF+Sham, or HF+BSG surgery. We used intention to treat analysis to evaluate VT/VF-free survival. We analyzed continuous ECG (VT/VF burden, heart rate variability, QT variability) and echo (cardiac function). In isolated left ventricular (LV) myocytes, we measured Ca 2+ transients and mitochondrial reactive oxygen species (mROS) using targeted ratiometric probes. Results: Half of the HF+Sham animals experienced SCD in 4 weeks. In contrast, BSG abolished SCD, reduced VT/VF, and increased parasympathetic tone (beyond corresponding reductions in sympathetic tone). Fractional shortening of the LV was markedly reduced in HF+Sham (0.28±0.02) vs. control (0.50±0.01), but preserved by BSG (0.45±0.02). Whereas HF+Sham increased mROS levels in LV myocytes (fractional oxidation, F ox : 0.53±0.015) compared to control (0.29±0.020), BSG reduced mROS (0.38±0.035). Mitochondrial antioxidant capacity, as indexed by H 2 O 2 challenge of LV myocytes, was compromised in HF+Sham (F ox : 0.81±0.040) compared to control (0.44±0.061), but preserved by BSG (0.48±0.023). Conclusions: Surgical BSG prevents SCD in non-ischemic HF by reducing mROS, decreasing sympathetic tone, and by increasing parasympathetic tone (possibly by facilitating cholinergic transdifferentiation). This promising new minimally-invasive surgical therapy may be particularly useful in patients with non-ischemic HF on optimal medical therapy who require recurrent ICD shocks to prevent SCD. Patients in the early stages of HF who are not eligible for ICDs but remain at higher SCD risk may also benefit from BSG surgery.

The Hemodynamic Response to Blood Loss in the Conscious Rat

The hemodynamic response to progressive blood loss passes through three distinct phases: an initial normotensive compensatory phase, a secondary hypotensive decompensatory phase, and a posthemorrhage recompensatory phase. The role of cardiac vagal and cardiac spinal signals in triggering the different phases of the response to hemorrhage was evaluated in the unanesthetized, freely moving rat by observing the effects on the response to 30% blood loss of prior cardiac vagal deafferentation (bilateral vagal rhizotomy) or prior cardiac spinal deafferentation (bilateral stellate ganglionectomy). In comparison to control animals, it was found that (i) cardiac spinal deafferentation significantly delayed the onset of the decompensatory phase, and (ii) cardiac vagal deafferentation slightly potentiated the decompensatory phase and impaired the recompensatory phase. These results indicate that it is cardiac spinal signals, rather than cardiac vagal signals, which in the conscious rat contribute to the triggering and progression of the decompensatory response to blood loss.

The effect of cardiac sympathetic denervation through bilateral stellate ganglionectomy on electrical properties of the heart

The role of the cardiac sympathetic nerve activity in various cardiac diseases is typically evaluated using β-adrenergic receptor antagonists. However, these antagonists induce global denervation effects not only in the cardiovascular system, but also in the brain and kidney. The objective of this study was to detect the electrophysiological property changes due to 8 days of cardiac sympathetic denervation and investigate the possible mechanisms underlying these changes using a more cardiac-specific bilateral stellate ganglionectomy (SGX) rat model. High-resolution optical mapping using a voltage-sensitive dye was performed in isolated Langendorff-perfused sham and SGX hearts, which were paced at progressively reduced basic cycle lengths under several different conditions: control, pretreatment with isoproterenol, and administration of atenolol and esmolol. Several electrophysiological parameters were recorded during periodic pacing and ventricular fibrillation (VF). Our results demonstrate that cardiac sympathetic denervation by bilateral SGX shortens action potential duration (APD) and flattens the APD restitution curve, but does not significantly affect spatial dispersion of APD. We found that, although the vulnerability of sham and SGX hearts to VF is similar, the dynamics of VF are different. The maximum dominant frequency is higher, and the spatial distribution of VF is more complex in the SGX heart, resulting in different mechanisms of VF. We demonstrated that β(1)-adrenergic receptors are downregulated in the SGX compared with sham hearts. In addition, our data suggest that the mechanism of cardiac sympathetic denervation by SGX surgery is more similar to the administration of β-blocker esmolol than atenolol.

Inhibition of Cardiac Ca 2+ Release Channels (RyR2) Determines Efficacy of Class I Antiarrhythmic Drugs in Catecholaminergic Polymorphic Ventricular Tachycardia

Catecholaminergic polymorphic ventricular tachycardia (CPVT) is caused by mutations in the cardiac ryanodine receptor (RyR2) or calsequestrin (Casq2) and can be difficult to treat. The class Ic antiarrhythmic drug flecainide blocks RyR2 channels and prevents CPVT in mice and humans. It is not known whether other class I antiarrhythmic drugs also block RyR2 channels and to what extent RyR2 channel inhibition contributes to antiarrhythmic efficacy in CPVT. We first measured the effect of all class I antiarrhythmic drugs marketed in the United States (quinidine, procainamide, disopyramide, lidocaine, mexiletine, flecainide, and propafenone) on single RyR2 channels incorporated into lipid bilayers. Only flecainide and propafenone inhibited RyR2 channels, with the S-enantiomer of propafenone having a significantly lower potency than R-propafenone or flecainide. In Casq2(-/-) myocytes, the propafenone enantiomers and flecainide significantly reduced arrhythmogenic Ca(2+) waves at clinically relevant concentrations, whereas Na(+) channel inhibitors without RyR2 blocking properties did not. In Casq2(-/-) mice, 5 mg/kg R-propafenone or 20 mg/kg S-propafenone prevented exercise-induced CPVT, whereas procainamide (20 mg/kg) or lidocaine (20 mg/kg) were ineffective (n=5 to 9 mice, P<0.05). QRS duration was not significantly different, indicating a similar degree of Na(+) channel inhibition. Clinically, propafenone (900 mg/d) prevented ICD shocks in a 22-year-old CPVT patient who had been refractory to maximal standard drug therapy and bilateral stellate ganglionectomy. RyR2 cardiac Ca(2+) release channel inhibition appears to determine efficacy of class I drugs for the prevention of CPVT in Casq2(-/-) mice. Propafenone may be an alternative to flecainide for CPVT patients symptomatic on β-blockers.

Effect of stellate ganglionectomy on basal cardiovascular function and responses to β1-adrenoceptor blockade in the rat

Cardiac sympathetic nerve activity is an important short-term controller of cardiac function and arterial pressure. Studies also suggest that long-term increases in cardiac sympathetic nerve activity may contribute to hypertension, coronary artery disease, and cardiac remodeling in heart failure. However, our understanding of the role of cardiac sympathetic nerves in chronic models of cardiovascular disease has been limited by inadequate experimental approaches. The present study was conducted to develop a surgical method to surgically denervate the sympathetic nerves of the rat heart for long-term cardiovascular studies. We characterized the effect of cardiac sympathetic denervation on basal levels of mean arterial pressure (MAP) and heart rate (HR) and the responses to a chronic administration of atenolol, a beta1-adrenoceptor antagonist. Rats were instrumented with telemetry transmitters for continuous recording of MAP and HR. After a 4-day baseline period, the rats were subjected to bilateral stellate ganglionectomy (SGX; n=9) or sham surgery (Sham; n=8). Seven days following SGX or Sham, the rats were administered atenolol for 5 days, followed by a 7-day recovery period. Following a transient decrease, SGX had no effect on basal MAP but decreased HR compared with baseline and Sham rats. Five days of atenolol treatment decreased MAP similarly in SGX and Sham rats. Atenolol resulted in a marked bradycardia in Sham rats but had a neglible effects on HR in SGX rats. The measurement of the content of cardiac catecholamines in all cardiac chambers at the end of the study verified a successful sympathetic denervation. This study confirms that bilateral SGX is a useful method to study the contribution of cardiac sympathetic nerves on the regulation of cardiac function. Moreover, these results suggest that cardiac sympathetic nerves are relatively unimportant in maintaining the basal level of MAP or the depressor response to atenolol in conscious, unrestrained rats.

Analysis of Cardiac Norepinephrine (NE) Transporter (NET) mRNA after removal of the stellate ganglia.

NET is an important protein in the regulation of catecholamines, particularly NE, and its dysfunction has been linked to certain cardiovascular diseases. To evaluate the contribution of the sympathetic innervation from the stellate ganglia as a source of NET in the heart, we compared by qPCR NET mRNA in the heart in control rats to rats after removal of stellate ganglia. NET mRNA was present in heart homogenate, with atria expressing more NET mRNA than ventricles (p&lt;0.05, n=4–6). Adult, male rats underwent bilateral stellate ganglionectomy (SGX) or sham surgery and hearts were assayed by chamber two weeks later. To confirm successful denervation, NE content was measured by HPLC and was reduced uniformly by 85.96% in all heart chambers (p&lt;0.001, n=4–5). Although NE was depleted, there was a significant increase in NET mRNA (p&lt;0.05, n=5) in the left atria while no differences were seen in any other chambers (p&gt;0.05, n=4–6). The presence of NET mRNA after SGX suggests that NET mRNA is present in non‐stellate sources and is regionally increased by removal of the sympathetic innervation. P01HL79687, AHA predoctoral fellowship (EW).

Ouabain-induced coronary vasoconstriction in cats is not neurally mediated.

1. Previous reports indirectly implicate a neural mechanism for coronary constriction to centrally administered digitalis. However, autoregulatory changes in coronary resistance due to changes in arterial pressure may have influenced the interpretation of these studies. 2. We tested directly the hypothesis that cardiac sympathetic innervation is responsible for coronary constriction to ouabain by examining the effects of ouabain (intravenous (i.v.) and intracerebroventricular (i.c.v.)) before and after bilateral stellate ganglionectomy. 3. Cats were anaesthetized and instrumented for the measurement of heart rate, blood pressure and coronary blood flow velocity using an epicardial-attached suction Doppler probe. Animals were treated with atenolol and the effects of either i.v. or i.c.v. injections of ouabain were examined. 4. In seven cats treated with atenolol, i.v. ouabain (0.11 mg/kg) produced maximal increases in arterial pressure and coronary vascular resistance index (CVRI) of 66 +/- 7 mmHg and 37 +/- 9%, respectively. Following bilateral stellate ganglionectomy (n = 7), ouabain produced similar increases in arterial pressure (70 +/- 9 mmHg) and CVRI (39 +/- 7%). A higher dose of i.v. ouabain (1.1 mg/kg) produced maximal increases in arterial pressure (115 +/- 4 mmHg) and coronary resistance (86 +/- 14%) in intact cats (n = 6) that were similar to responses seen in cats in which stellate ganglionectomy had been performed (n = 6; arterial pressure 104 +/- 13 mmHg; coronary resistance 114 +/- 6%). The increases in coronary resistance to ouabain at both doses were significantly greater than increases in coronary resistance to passive elevation of arterial pressure during aortic constriction. Thus, autoregulation does not explain fully the coronary constriction to ouabain. 5. To further examine a central mechanism, i.c.v. perfusion with 0.3 mmol/L ouabain was performed in six cats, resulting in increases in arterial pressure (122 +/- 7 mmHg) and coronary resistance (58 +/- 14%). Similar increases in arterial pressure (117 +/- 16%) and coronary resistance (84 +/- 20%) were seen in separate studies (n = 6) following stellate ganglionectomy. 6. These results indicate that coronary constriction to ouabain does not require intact cardiac sympathetic innervation, but probably involves a direct or humorally mediated effect.

Age-related effects of cardiac sympathetic denervation on the responses to cardiopulmonary receptor stimulation in piglets.

The effects of right stellate ganglionectomy (RSG) and bilateral stellate ganglionectomy (BSG) on cardiovascular responses to phenyl biguanide (PBG, 80 micrograms/kg) were studied in 1- and 8-wk-old piglets. Animals were anesthetized with Saffan, paralyzed, thoractomized, and ventilated with 100% O2. Recordings of the ECG (lead II) and aortic pressure (AoP) were used to compute the maximum R-R interval, heart rate (HR), and mean AoP, and to determine the occurrence of atrioventricular conduction block (AVB). Right atrial injections of PBG in 1-wk-old piglets elicited AVB as well as decreases in AoP and HR in all animals; this response pattern was not altered by either RSG or BSG. The PBG response of neurally intact 8-wk-old animals was comprised of a decrease of HR without change in AoP; AVB occurred in three of six animals. After RSG or BSG, AoP decreased along with decreased HR, and now AVB occurred in all animals; changes of AoP and maximum R-R interval were greater after BSG than after RSG. These results suggest that the stellate ganglia exert a neuroprotective influence on cardiovascular function, requiring some degree of maturation for expression. Our findings support the hypothesis that an imbalance of cardiac autonomic innervation favoring parasympathetic activity may produce immature responses to cardiopulmonary afferent stimulation in older maturing animals.

THE EFFECTS OF SELECTIVE CARDIAC SYMPATHETIC DENERVATION ON VENTRICULAR FIBRILLATION THRESHOLD (VFT) IN DEVELOPING SWINE • 215

Abnormal cardiac autonomic innervation may be a mechanism leading to sudden infant death syndrome (SIDS). The present study evaluated the influence of selected cardiac denervations on ventricular fibrillation threshold (VFT) in one month old swine. Saffan-anesthetized, open-chest swine were artificially ventilated with 100% O2. Aortic pressure (AoP) and ECG (Lead II) were recorded. For VFT measurement, a train of 10 pulses of 10 msec duration at 20 Hz was delivered through bipolar electrodes positioned at the apex of the heart. Current intensity was progressively increased by 0.5 mA until ventricular fibrillation occurred. The lowest current intensity, which induced fibrillation, was defined as VFT. Defibrillation was accomplished within 15 sec following fibrillation. The VFT measurement was repeated 30 min later following recovery of the ECG, AoP, blood gases and pH. In sham control: 1st VFT= 4.8 ± 1.0 mA vs 2nd VFT = 4.4 ± 1.6 mA, Mean ± SD, n=5, p>0.05. For the denervation groups (with vagi intact): VFT was measured pre- and 30 min post right stellate ganglionectomy (RSG) or left stellate ganglionectomy (LSG) or bilateral stellate ganglionectomy (BSG). There were significant decreases in VFT following RSG (5.0 ± 1.3 vs 2.9± 1.3 mA, p 0.05) or BSG (5.30 ± 1.8 vs 5.24± 1.9 mA, n=8, p>0.05). The results suggest that elimination of the right stellate (left-sided dominance) increase susceptibility to ventricular fibrillation while elimination of the left or both stellates in the swine did not alter VFT. Thus we speculate that inappropriate development of cardiac autonomic innervation in the neonate may be a mechanism in the etiology of SIDS.(Supported by NIH grant HD-28931).

Parasympathetic control of cycle length dependence of endocardial ventricular repolarisation in the intact feline heart during steady state conditions

Parasympathetic modulation of the rate dependence of ventricular repolarisation in vivo during steady state conditions was investigated by analysing the effects of removing vagal activity in three groups of anaesthetised cats. Bilateral vagotomy was performed while different levels of background sympathetic tone were present in the control periods of each group: in group 1 (n = 6), cardiac sympathetic nerves were intact; in group 2 (n = 7), bilateral stellate ganglionectomy was performed; and in group 3 (n = 7), beta adrenergic blockade (propranolol, 0.5 mg.kg-1) was performed after stellate ganglionectomy. The duration of a left ventricular endocardial monophasic action potential (APD) was measured during atrial pacing at 7-10 cycle lengths (CL). The APD/CL relation was fitted to a hyperbolic function: APD = CL/[(axCL)+b]. Two parameters were considered: APDmax (1/a, ie, APD extrapolated at infinite cycle length, a rate-independent measure of APD) and CL50 (bxAPDmax, ie, the cycle length at which 50% of APDmax is reached). In control conditions, APDmax and CL50 were longer in groups 2 and 3, when cardiac sympathetic effects were reduced or absent. Vagotomy reduced APDmax and CL50 similarly in groups 1 and 2 (APDmax, -24% and -18%; CL50, -36% and -27%) (p < 0.05 v control). In group 3, vagotomy did not affect APDmax and CL50. No direct parasympathetic influence on the rate dependence of endocardial ventricular repolarisation was observed. The vagal modulation of sympathetic effects may take place either through vago-sympathetic reflexes or via the antagonism of circulating catecholamines distal to the beta receptor.

Bradykinin increases myocardial contractility: relation to the Gregg phenomenon

Bradykinin (BK) is reportedly produced in the heart during ischemia. Because BK has been shown to activate cardiac afferent nerves thought to be nocioceptors, we tested whether BK might alter myocardial shortening, which potentially could contribute to afferent nerve stimulation. In open-chest dogs, BK (1-10 micrograms) was injected into the left anterior descending (LAD) coronary artery while wall motion in the LAD and control circumflex regions was monitored. Wall motion was measured with midwall segment gauges (sonomicrometer crystals) placed in the hoop direction. Blood pressure, heart rate, left ventricular pressure, first derivative of left ventricular pressure, and LAD coronary flow also were monitored. At 15-20 s after injection, which was before circulation of the peptide caused blood pressure to change, BK decreased maximum end-diastolic and minimum end-systolic segment lengths and increased maximum shortening fraction in LAD region. No change was observed in circumflex region. The response was not eliminated by bilateral vagotomy or subsequent stellate ganglionectomy, indicating that it was not neurally mediated. The response closely paralleled changes in coronary flow, was mimicked by intracoronary injection of adenosine, and was reduced or absent if flow was already elevated by previous injection of adenosine. When BK eventually reached the systemic circulation, the resultant hypotension further reduced shortening in LAD region, with directionally similar effect in circumflex region. These results suggest that BK can increase regional shortening by enhancing coronary flow (Gregg phenomenon) as well as by altering global ventricular function through systemic hypotension. Such changes in shortening may contribute to stimulation of cardiac afferent nerves.

Stimulation of serotonin2 receptors in the ventrolateral medulla of the cat results in nonuniform increases in sympathetic outflow.

Topical application of the serotonin2 agonist, 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane or DOI, in a dose of 30 micrograms/side to the intermediate area of the ventrolateral surface of the medulla produced a significant increase in mean arterial pressure with no significant change in heart rate both in intact animals (n = 8) and in cervically vagotomized animals (n = 3). The pressor response of DOI was blocked by pretreatment of the intermediate area with ketanserin, a serotonin2 antagonist (n = 7). Pretreatment with intravenous phentolamine did not block the pressor response of DOI (n = 3). However, this pressor response could be counteracted by intravenous propranolol (n = 5) or by bilateral stellate ganglionectomy (n = 3). These data suggest that sympathoexcitation by centrally applied DOI selectively increased cardiac inotropy but not chronotropy. Further studies indicate that DOI increased contractile force without increasing heart rate and that the positive inotropic effect of DOI could be counteracted by bilateral stellate ganglionectomy. Bilateral microinjections of DOI into the subretrofacial nucleus in a dose of 100 ng (n = 3) and a dose of 300 ng (n = 3) increased mean arterial blood pressure by 23 +/- 2 and 44 +/- 6 mm Hg, respectively, without producing any changes in heart rate. These data suggest that DOI has a central site of action in the ventrolateral medulla, presumably at the subretrofacial nucleus, which leads to selective sympathoexcitation of the cardiac ventricles.

Antifibrillatory action of the narcotic agonist fentanyl

Morphine, an opiate alkaloid with mixed mu- and delta-agonist properties, raises the ventricular fibrillation threshold in anesthetized dogs by altering autonomic tone. To elucidate further underlying structure-activity relationships, the effect of fentanyl, a nonalkaloid, mu-selective agonist in wide clinical use, was studied. Fentanyl (30 μg/kg) was given intravenously to 27 chloralose-anesthetized dogs, and ventricular fibrillation threshold was measured by means of the single-stimulus technique. In the baseline state fentanyl raised the ventricular fibrillation threshold by 14%. When the dogs were subjected to hemorrhagic stress, this effect was amplified to 29% ( p < 0.0001). Bilateral cervical vagotomy abolished fentanyl's antifibrillatory effect, but neither atropine sulfate (0.4 mg/kg/hr) nor atropine methylnitrate (0.5 mg/kg/hr) did so. Fentanyl's influence on the fibrillation threshold during hemorrhage was significantly reduced by bilateral stellate ganglionectomy ( p < 0.005). It is concluded that fentanyl raises the ventricular fibrillation threshold by its known sympathoinhibitory action rather than by its vagal efferent activating effect. The facts that an intact vagus is required and that hemorrhage amplifies the effect suggest that the antifibrillatory effect of fentanyl is mediated through the afferent component of the baroreflex arc.

Autonomie control of ventricular tachycardia. III. Effects of adenosine and N6-R-1-phenyl-2-propyladenosine

The purpose of this study was to determine whether adenosine or the adenosine deaminase-resistant analogue, N6-R-1-phenyl-2-propyladenosine (RPIA), could slow the rate of spontaneous ventricular tachycardia occurring 24 hours after left anterior descending coronary artery occlusion. Chloralose-anesthetized, open chest dogs (n = 25) with ventricular tachycardia were studied. The left anterior descending artery was cannulated distally. Intracoronary infusions of adenosine, 10(-7) to 10(-5) M, did not alter the rate of ventricular tachycardia. Ventricular tachycardia slowed by 4.6% with adenosine, 10(-4) M. RPIA, 10(-6) to 10(-4) M, produced a concentration-dependent decrease in the rate of ventricular tachycardia when injected into the left anterior descending coronary artery. This effect of RPIA was reversed by the adenosine antagonist aminophylline, 10(-5) M. After bilateral stellate ganglionectomy, RPIA, 10(-5) M, did not, but metoprolol, 0.5 mg, did slow ventricular tachycardia after intracoronary injection. However, RPIA, 10(-5) M, produced a 43% decrease in the increment in ventricular tachycardia occurring during sympathetic neural stimulation. Therefore, when injected into the left anterior descending artery, adenosine, 10(-4) M, and RPIA, 10(-6) to 10(-4) M, decrease the rate of ventricular tachycardia in 24 hour old myocardial infarction. Furthermore, this decrease in the rate of ventricular tachycardia is the result of prejunctional sympathetic antagonism.

The effects of elevated intracranial pressure on the canine electrocardiogram

Striking electrocardiographic abnormalities have been noted in some patients with central nervous system injury. To study the relationship between the electrocardiogram and intracranial pressure, intracranial pressure was elevated in 14 open chest pentobarbital-anesthetized dogs. The right vagus was stimulated to produce sinus slowing and the right atrium was paced at a constant cycle length fast enough to prevent arrhythmias and maintain heart rate constant (750 msec in 11 dogs and 600 msec in three dogs). In nine dogs, intracranial pressure was sequentially elevated to 100, 150, and 200 mmHg. Systolic arterial blood pressure consistently rose to exceed intracranial pressure (P less than 0.005). At a pressure of 150 and 200 mmHg, mean QT intervals shortened significantly in recorded leads II, X, Y, and Z from 0.01). T wave changes were also noted that consisted of increasing positivity in leads II, X, and Y and increasing negativity in lead Z. To delinate the role of the sympathetic nervous system, an additional five dogs were subjected to an intracranial pressure of 200 mmHg before and after bilateral stellate ganglionectomy and timolol (0.1 mg/kg IV). Elimination of sympathetic influences did not significantly alter the electrocardiographic effects of elevated intracranial pressure. Thus, intracranial hypertension results in significant QT shortening and T wave changes that are not entirely mediated by the sympathetic nervous system.

Role of Myocardial Hypertrophy in Trophic Stimulation of Indices of Sympathetic Cardiac Innervation

Summary: Indices of cardiac sympathetic innervation have commonly been found depressed in the failing, hypertrophied heart. In contrast, we have recently demonstrated that hemodynamically compensated, very gradually developing right ventricular hypertrophy is associated with an increase in sympathetic nervous markers. The present experiments were performed to corroborate these findings in a model of acutely induced right ventricular hypertrophy, and to further characterize changes in markers of autonomic innervation associated with cardiac hypertrophy. Male guinea pigs underwent either pulmonary artery banding (P) with an acutely constricting ligature, or bilateral stellate ganglionectomy (S), or both (PS). Appropriate sham procedures were performed in animals subjected to only one intervention; controls (C) underwent sham-S and sham-P. Groups of animals were sacrificed at 10 and 20 days after surgery. Cardiac tissues were weighed and subsequently analyzed for activities of tyrosine hydroxylase (TH) and dopamine β-hydroxylase (DBH), two enzymes catalyzing the biosynthesis of catecholamines (CAs), and of choline acetyltransferase (CAT), a marker of parasympathetic activity, as well as for norepinephrine (NE). S resulted in profound depletions of cardiac NE of 88-92% and in significant decreases in the activities of DBH and TH. Marked right ventricular hypertrophy developed rapidly following P, and was not modified by S. Similar to our previous results, acute right ventricular hypertrophy was associated with moderate increases (10-20%) of sympathetic markers; following S, these increases (of presumably residual sympathetic innervation) were greatly enhanced, amounting to 171% and 105% for NE at 10 and 20 days, respectively. In contrast, sympathetic markers in the left ventricle of stellatectomized animals were not affected by P. Activity of CAT remained unaltered by the experimental interventions. Our experiments indicate that increases in markers of sympathetic innervation may be a common feature of the early, compensated stage of cardiac hypertrophy, regardless of its time course. Sympathetic neural mechanisms do not appear to play a stimulatory or trophic role in the hypertrophic process. Conversely, they seem to be secondary in nature, suggesting a possible stimulatory influence of hypertrophying myocardium on sympathetic cardiac nerves.

Role of Myocardial Hypertrophy in Trophic Stimulation of Indices of Sympathetic Cardiac Innervation

Indices of cardiac sympathetic innervation have commonly been found depressed in the failing, hypertrophied heart. In contrast, we have recently demonstrated that hemodynamically compensated, very gradually developing right ventricular hypertrophy is associated with an increase in sympathetic nervous markers. The present experiments were performed to corroborate these findings in a model of acutely induced right ventricular hypertrophy, and to further characterize changes in markers of autonomic innervation associated with cardiac hypertrophy. Male guinea pigs underwent either pulmonary artery banding (P) with an acutely constricting ligature, or bilateral stellate ganglionectomy (S), or both (PS). Appropriate sham procedures were performed in animals subjected to only one intervention; controls (C) underwent sham-S and sham-P. Groups of animals were sacrificed at 10 and 20 days after surgery. Cardiac tissues were weighed and subsequently analyzed for activities of tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH), two enzymes catalyzing the biosynthesis of catecholamines (CAs), and of choline acetyltransferase (CAT), a marker of parasympathetic activity, as well as for norepinephrine (NE). S resulted in profound depletions of cardiac NE of 88-92% and in significant decreases in the activities of DBH and TH. Marked right ventricular hypertrophy developed rapidly following P, and was not modified by S. Similar to our previous results, acute right ventricular hypertrophy was associated with moderate increases (10-20%) of sympathetic markers; following S, these increases (of presumably residual sympathetic innervation) were greatly enhanced, amounting to 171% and 105% for NE at 10 and 20 days, respectively. In contrast, sympathetic markers in the left ventricle of stellatectomized animals were not affected by P. Activity of CAT remained unaltered by the experimental interventions. Our experiments indicate that increases in markers of sympathetic innervation may be a common feature of the early, compensated stage of cardiac hypertrophy, regardless of its time course. Sympathetic neural mechanisms do not appear to play a stimulatory or trophic role in the hypertrophic process. Conversely, they seem to be secondary in nature, suggesting a possible stimulatory influence of hypertrophying myocardium on sympathetic cardiac nerves.

Respiratory responses to chemical activation of left ventricular receptors

Respiratory responses to chemical activation of receptors in the left ventricle were measured in anesthetized cats. Application of capsaicin (10 μg) or bradykinin (500 ng) to the serosal surface of the left ventricle caused increases in phrenic nerve activity and in arterial pressyre. These responses persisted after cervical vagotomy and after bilateral transection of the carotid sinus nerves. Bilateral stellate ganglionectomy abolished the respiratory responses to capsaicin and bradykinin. In addition, stellate ganglionectomy prevented the arterial pressure response to bradykinin; however, a slight increase in arterial pressure occurred with application of capsaicin. It is concluded that activation of sympathetic afferents orginating in the left ventricle caused an increase in respiratory output.