Cardiovascular effects of atrial natriuretic extract in the whole animal

Atrial tissue extract (AE) and ventricular tissue extract cause identical decreases in total peripheral resistance when they are injected i.v. into anesthetized rats. However, only AE causes significant hypotension because of cardiac inhibition. This involves both bradycardia and failure of stroke volume to increase appropriately. The observations cannot be explained by direct action of AE on myocytes, but are more likely to be the result of interactions with cardiovascular reflex mechanisms. Excitation of chemosensitive cardiac receptors with vagal afferents appears to be an important afferent mechanism. The efferent limb for the negative chronotropic response resides partly in the vagus nerves and partly in cardiac sympathetic nerves. The negative inotropic response of AE was not altered by vagotomy, spinal section, atropine, or propranolol. These results suggest that atrial peptides may cause the release of a negatively inotropic substance from a site that is not yet identified.     

Pharmacological evidence for cardiac muscarinic receptor subtypes

The chronotropic and inotropic effects of muscarinic receptor agonists (Acetylcholine, Arecoline, Carbachol, Furtrethonium) and antagonists (Atropine, N-methyl and N-butyl scopolammonium, pirenzepine) on isolated guinea-pig atria were studied. All had a greater affinity constants for muscarinic receptors as assessed in terms of inotropic effects than in terms of chronotropic effects. This difference, well correlated with the pharmacological effect, suggests the occurrence of cardiac muscarinic receptor subtypes, one mediating heart rate and the other contractile force. The ratio of chronotropic to inotropic potencies for each agent shows that the physiological mediator. Acetylcholine, differentiates best between the two subtypes, while atropine is the least discriminatory.     

The structural dynamics of vagus influence on the heart rhythm in exposures directed at altering the acting acetylcholine concentration]

In 29 experiments on anaesthetized cats burst stimulation of peripheral cut end of right vagus nerve leads to synchronization of heart and vagus rhythm. Influence of proserine, pilocarpine and prolonged vagus stimulation upon extent of vagus chronotropic effect and its components--tonic and synchronizing--was investigated. In all cases changes of vagus chronotropic effect during this actions were caused by unidirectional shifts of tonic component. Extent of synchronizing vagus chronotropic influences did not depend on the changes of acetylcholine concentration.     

Selective pharmacological blocking of synaptic transmission through the parasympathetic ganglia: Effects on the cardiovascular system

In acute experiments on rats and dogs, compounds IEM-1556 and IEM-1678, the blockers of transmission through the parasympathetic ganglia, reduced the negative chronotropic effect of stimulation of the vagus nerve (VN), while practically not changing the heart rate (HR). In chronic experiments on dogs, these compounds increased the HR, substantially reduced the respiratory heart arrhythmia, did not change the arterial blood pressure (AP), and reduced the chronotropic effects of VN stimulation. IEM-1556 exerted more strong and long-lasting blocking effects on vagal heart control than IEM-1678 did, but in anesthetized animals could evoke a drop in the AP. Acetylcholine, if administered during the action of the above compounds, inhibited heart activity. It is concluded that both IEM-1678 and IEM-1556 are selective parasympatholytics (although IEM-1556 may produce a side effect). The above compounds block synaptic transmission through the intracardiac parasympathetic ganglia and do not affect neuro-effector transmission in the heart.Neirofiziologiya/Neurophysiology, Vol. 28, No. 2/3, pp. 151–159, March–June, 1996.     

Mechanism for amine modulation of the neurogenic Limulus heart: evidence for involvement of cAMP

The role of cyclic nucleotides as intracellular second messengers mediating the excitatory chronotropic and inotropic actions of octopamine (OCT) and dopamine (DA) on the neurogenic Limulus heart was investigated. Tissue levels of cAMP, but not cGMP, were significantly increased in isolated cardiac ganglia and cardiac muscle following 10 min exposure to 10(-5) M OCT or 10(-5) M DA. In both tissues, OCT elicited larger increases in cAMP than did DA. Amine-induced cAMP accumulation in the cardiac ganglion and in the cardiac muscle was prevented by the alpha-adrenergic blocker phentolamine. The adenylate cyclase activator forskolin and the phosphodiesterase inhibitor IBMX produced amine-like chronotropic and inotropic effects when applied to the isolated heart preparation. However, the kinetics of the responses differed for the two agents. Additional pharmacological agents (RO-20-1724, papaverine, SQ 20,009, and 8-parachloro-phenylthio cAMP) also had amine-like effects but to a lesser extent. The chronotropic, but not inotropic, effects of OCT and DA were potentiated in the presence of IBMX. These data suggest that a cAMP-dependent mechanism underlies the excitatory effects of the neuromodulators OCT and DA on the Limulus heart.     

The action of parotoid venom on the heart of the toad (Bufo ictericus ictericus Spix 1824) and its effects on the inhibition caused by vagal stimulation

1. The administration of crude venom of the parotoid glands of the toad Bufo ictericus ictericus to the in situ (via abdominal vein) or isolated heart of this anuran causes both chronotropic and inotropic effects. 2. While under action of parotoid venom, the heart of the animal is insensitive to vagus nerve stimulation. 3. This blocking of vagal action is dose dependent and it is suggested that it results from a functional antagonism between the venom constituents and the acetylcholine liberated by the nerve endings on stimulation. 4. The venom constituents probably involved in this antagonism are catecholamines (adrenaline and noradrenaline), tryptamine derivatives (serotonin and bufotenidin) and genins (bufagin and bufotoxin), possibly also ATP. 5. Adrenaline, noradrenaline and serotonin, or a mixture of the three, mimic, at least partially, the blocking of vagal action caused by crude venom. 6. The blocking action of crude venom can be prevented by previously or simultaneously adding acetylcholine to the infused crude venom. This prevention is dose dependent. 7. The blocking action persists in the boiled venom and in the material dialysed from crude venom.     

Fade of the responses of the isolated, blood-perfused dog atrium to cholinergic interventions

In the isolated, blood-perfused, canine right atrium, intramural parasympathetic nerve stimulation and intra-arterial infusions of acetylcholine induced substantial negative chronotropic and inotropic responses. The responses to parasympathetic stimulation reached their maximum values quickly, and then usually faded back toward control levels over the next 1 or 2 min of stimulation. The fade of the responses at high stimulation frequencies (greater than or equal to 30 Hz) was significantly greater than that at lower frequencies. The inotropic responses to acetylcholine infusion (1 microgram/min) faded slightly but significantly, whereas the chronotropic responses did not fade at all. These results suggest that the fade of the cardiac responses to parasympathetic stimulation is mainly ascribable to a progressive reduction in the rate of acetylcholine release from the nerve endings, especially at higher stimulation frequencies. The fade of the inotropic responses was more pronounced and had a longer time course than that of the chronotropic responses. Furthermore, the fade of the inotropic responses diminished significantly as the response magnitude was augmented by an increase in stimulation voltage. Conversely, the fade of chronotropic responses was not significantly affected by this intervention. These differences in the inotropic and chronotropic responses to neural stimulation, and the occurrence of a slight fade of the inotropic response to acetylcholine infusion, suggest that in addition to the predominant prejunctional mechanism, a postjunctional phenomenon may also be partly responsible for the fade of the inotropic response to cholinergic interventions.     

Hemodynamic changes during electrical stimulation of some bulbar cardiovascular structures

The effect of electrical stimulation was studied on the nucleus of the tractus solitarius, the nucleus cuneatus, and the dorsal motor nucleus of the vagus nerve, i.e., structures of the bulbar cardiovascular sector in which it has been postulated that impulses from the sinoaortic reflexogenic zone are relayed to the cardiovascular system. Stimulation of all the structures tested in acute experiments on cats under chloralose-Nembutal anesthesia evoked depressor responses of varied degree, the hemodynamic basis of which was a decrease in the cardiac output (CO). The effect of stimulation of the nucleus cuneatus on the development of the negative chronotropic effect was observed. The dorsal motor nucleus of the vagus nerve in cats is not the only or even the principal zone from which negative chronotropic influences are exerted on the heart.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol.3, No.6, pp.631–636, November–December, 1971.     

The spectrogram of heart rate in cats following burst stimulation of the vagus nerve

Denervation of the heart (bilateral vagotomy and propranolol) in artificially ventilated cats didn't remove respiratory peaks on the spectrogram of heart rate, while burst stimulation of vagus nerve increased or decreased them several times by synchronization of the heart and vagus rhythms, which in its turn was observed under the bradycardia only. At the same time, the desynchronization of rhythms provoked severe sinus arrhythmia which had a distinct periodic character. Under these conditions, there were high non-respiratory peaks appearing at the spectrogram of the heart rate that indicated existence of two vagus chronotropic effects: a well known tonic one and special intracycle synchronizing effect correcting duration of every cardiac cycle.     

Developmental changes in dopamine modulation of the heart in the isopod crustacean Ligia exotica: reversal of chronotropic effect

Developmental changes in dopamine modulation of the heart were examined in the isopod crustacean Ligia exotica. The Ligia cardiac pacemaker is transferred from the myocardium to the cardiac ganglion during juvenile development and the heartbeat changes from myogenic to neurogenic. In the myogenic heart of early juveniles, dopamine affected the myocardium and caused a decrease in the frequency and an increase in the duration of the myocardial action potential, resulting in negative chronotropic (decrease in beat frequency) and positive inotropic (increase in contractile force) effects on the heart. Contrastingly, in the heart of immature adults just after juvenile development, dopamine caused effects of adult type, positive chronotropic and positive inotropic effects on the heart affecting the cardiac ganglion and myocardium. During the middle and late juvenile stages, dopamine caused individually a negative or a positive chronotropic effect on the heart. These results suggest that the chronotropic effect of dopamine on the Ligia heart is reversed from negative to positive in association with the cardiac pacemaker transfer from the myocardium to the cardiac ganglion during juvenile development.     

Pituitary adenylate cyclase-activating polypeptide: localization and differential influence on isolated hearts from rats and guinea pigs

This study was done to determine if pituitary adenylate cyclase-activating peptide (PACAP)-immunoreactive nerve fibers occur in cardiac muscle as well as intracardiac ganglia of rats and guinea pigs and to clarify the chronotropic actions of PACAP27 in the same species using isolated heart preparations. PACAP nerve fibers were not detected in atrial or ventricular muscle of rat or guinea pig but a few stained nerve fibers occurred in the atrioventricular bundle of the guinea pig. Stained nerve fibers were prominent in intracardiac ganglia of both species. PACAP27 caused a dose-dependent tachycardia in isolated rat hearts (+39 +/- 3 beats/min with 1 nmol, n = 6). Positive and/or negative chronotropic responses were evoked by PACAP27 in guinea pig heart, depending on dose and prior exposure to the peptide. PACAP27 also caused arrhythmias in several guinea pig hearts. Treatment with atropine eliminated or prevented PACAP-evoked bradycardia and arrhythmias, implicating cholinergic neurons in these responses. Positive chronotropic responses to PACAP were unaffected by beta-adrenergic receptor blockade in either species, suggesting that tachycardia resulted from a direct action on the heart. These observations support the conclusion that endogenous PACAP could have a role in regulating parasympathetic input to the heart but through different mechanisms in rats versus guinea pigs. A direct positive chronotropic influence of endogenous PACAP is unlikely since atrial muscle lacks PACAP-immunoreactive nerve fibers.     

Regulation of atrial contraction in the seawater-adapted eel,Anguilla japonica

The atrium isolated from the seawater-adapted eel beats spontaneously in normal Ringer solution for more than 10 hr. The strength of beating was inhibited by acetylcholine (ACh) and the inhibitory effects were blocked by atropine, a muscarinic ACh-receptor antagonist, indicating existence of muscarinic ACh-receptor on the atrium. The atrial contractility was stimulated by catecholamines and their agonists; the order of potency being isoproterenol > adrenaline (AD) = noradrenaline (NA) > phenylephrine > clonidine. The stimulatory effects of AD was completely blocked by propranolol, a β-adrenoceptor antagonist, but not by phentolamine, an α-adrenoceptor antagonist. These data were consistent with characteristics of β-adrenoceptors. Further characterization of the β-receptor was not attempted. The positive inotropic and chronotropic actions of AD were not completely blocked either by atenolol, a β₁-adrenoceptor antagonist, or by ICI 118551, a β₂-adrenoceptor antagonist. When electrical current with a short duration (0.25 msec) was passed through the atrium, the beating was inhibited initially, then enhanced later. The initial inhibition was inhibited by atropine and the later enhancement was blocked by propranolol. These results indicate that the electrical stimulation releases ACh and catecholamine(s) from the nerve endings. The positive inotropic and chronotropic effects of catecholamines were mimicked by tyramine, a catecholamine releaser from sympathetic nerve endings.     

Chronic atropine administration diminishes the contribution of vasoactive intestinal polypeptide to heart rate regulation

Vasoactive intestinal polypeptide (VIP) is implicated in the modulation of vagal effects on the heart rate. In this study, the impact of acute and chronic atropine administration on VIP levels in rat heart atria was investigated in relation to heart rate in the course of vagus nerves stimulation. Anaesthetised control and atropinised (10 mg/kg/day for 10 days) rats pretreated with metipranolol and phentolamine that were either given or not a single dose of atropine were subjected to bilateral vagus nerve stimulation (30 min: 0.7 mA, 20 Hz, 0.2 ms). VIP concentrations in the atria were determined after each stimulation protocol. In control rats with or without single atropine administration, the heart rate upon vagal stimulation was higher than in atropinised animals with or without single atropine dose, respectively. VIP concentrations in the control atria were significantly decreased after the stimulation; the decrease was comparable both in the absence and presence of a single dose of atropine. Compared to controls, VIP levels were significantly decreased after chronic atropine treatment and they were not further reduced by vagal stimulation and single atropine administration. Administration of VIP antagonist completely abolished the differences in the heart rate upon vagal stimulation between control and atropinised groups. In conclusion, the data indicate that chronic atropine administration affects VIP synthesis in rat heart atria and consequently it modifies the heart rate regulation.     

Studies on bradycardia mechanism in the moderately hypothermic dog.

Chronotropic action of isoprenaline on the heart was studied in anesthetized dogs, in euthermic and moderate hypothermic conditions, before and after intravenous administration of atropine and oxprenolol or a cervical bilateral vagotomy. In moderate hypothermia we observed: i) larger duration of the positive chronotropic response to isoprenaline with a delayed and slightly lesser intensity in its maximum; ii) relating to euthermic conditions, delayed but superimposed potentiation of the chronotropic isoprenaline response in atropinized or vagotomized dogs; iii) a small negative chronotropic response to isoprenaline 15 min after oxprenolol, that diminished after atropine; iiii) oxprenolol induced a marked bradycardia nearly twice as intense as in euthermic dogs, almost completely blocked subsequently by atropine. It is concluded that progressive bradycardia in the moderately hypothermic dog is due, among other factors, to a cholinergic action but not to a lesser ability of beta-adrenergic cardiac effectors to chronotropic responses.     

Enhanced cAMP accumulation after termination of cholinergic action in the heart

Cholinergic agents decrease myocardial contractility in part by inhibiting adenylate cyclase (EC 4.6.1.1) activity. We have found that after a prolonged preincubation period (greater than 6 h), washout of cholinergic agents from embryonic chick hearts or cultured heart cells results in a persistent increase in their basal and catecholamine-stimulated cAMP content. Membranes prepared from pretreated cells have elevated basal, forskolin-, and catecholamine-stimulated adenylate cyclase activities. This myocardial adaptation to cholinergic agents is analogous to changes in nerve cells and other cell types after prolonged exposures to narcotics or other inhibitors of adenylate cyclase, respectively. A rapid (less than 5 min) adaptation response to cholinergic agents can also be demonstrated in heart cells by quickly blocking agonist action with atropine. Atropine alone has no effect, but after a brief preincubation period with agonists (methacholine or oxotremorine), the addition of atropine transiently enhances catecholamine-stimulated cAMP accumulation by 2.5-fold. These responses are absent in heart cells pretreated with pertussis toxin. The data indicate that the response is not mediated by the phosphoinositide pathway, which has been demonstrated to be insensitive to pertussis toxin in chick heart. Enhanced cAMP accumulation after termination of muscarinic agonist action may provide an explanation for the observation that acetylcholine sometimes produces biphasic contractile responses.     

Somatostatin as a modulator of vagal effects on heart rhythm]

In 11 experiments on anesthetised cats burst stimulation of peripheral cut end of right vagus nerve leads to synchronization of cardiac and vagus rhythms. Alterations of burst sequence frequency within definite limits has been synchronously reproduced by heart thus creating managed bradycardia possibility. Somatostatin (10(-8)-10(-9) M intravenously) decreases heart rate and inhibits total vagus chronotropic effect. Vagolytic effect of somatostatin caused a decrease of tonic component of the vagus chronotropic effect. On the other hand, somatostatin augmented the extent of the vagal synchronizing influences and caused enlargement of the ranges of managed bradycardia. The observed results testify to participation of the peptidergic mechanisms in genesis of vagal managed bradycardia.     

Alpha- and gamma-endorphin suppression of the chronotropic effects of exogenous acetylcholine

On the isolated frog heart alpha- and gamma-endorphins depressed the chronotropic effects of exogenous acetylcholine. Naloxone partially blocked the atropine-like action of alpha-, gamma-endorphins. The latent period of the reaction to the endorphin injection was about 20 min, the endorphin effect retained more than 1 h. The obtained data show that some opioids may act as "endogenous atropine".     

The roles of alpha- and beta-adrenoceptors in the chronotropic responses to norepinephrine in carp heart (Cyprinus carpio)

1. The chronotropic effect of norepinephrine was studied in isolated spontaneously beating atrial preparations of carp (Cyprinus carpio) heart. 2. Norepinephrine, 0.1 microM, caused a positive chronotropic effect, while at 1 microM it caused either a positive or a negative chronotropic effect. The positive chronotropic effect, observed in 13 preparations, was potentiated by phentolamine and almost completely blocked by propranolol. 3. The negative chronotropic effect observed in the other 5 preparations was greater in the presence of propranolol, reduced by phentolamine and not affected by atropine. 4. These results indicate that alpha- and beta-adrenoceptors may coexist, mediating the negative and positive chronotropic effects, respectively, in isolated atrial preparations of carp heart.     

The chronotropic action of neurotensin in the guinea pig isolated heart

Neurotensin (NT) infusions into isolated, perfused, spontaneously beating hearts of guinea pigs evoked a concentration-dependent, positive chronotropic effect which was preceded in some hearts by transient bradycardia. The tachycardia caused by NT was not affected by propranolol, cimetidine, indomethacin, a mixture of methysergide and morphine or by atria removal. The incidence and amplitude of bradycardia caused by NT were increased by neostigmine but reduced by atropine. Neostigmine and atropine also tended to decrease and increase respectively, the tachycardia caused by NT. These results suggest that the positive chronotropic effect of NT in guinea pig isolated heart results from a direct effect on the specialized conduction system of the heart while its negative chronotropic effect is likely to reflect the activation by NT of cardiac vagal cholinergic neurons.     

Capsaicin-sensitive structures as potential target sites for neurotensin and bradykinin in guinea pig atria

We tested the influence of capsaicin (CAP) desensitization on the positive chronotropic and inotropic effects of neurotensin (NT), bradykinin (BK), calcitonin gene-related peptide (CGRP) and noradrenaline (NA) in guinea pig isolated atria. The positive chronotropic and inotropic effects of NT and BK were completely inhibited, whereas those elicited by CGRP and NA were either slightly reduced (CGRP) or unaffected (NA), in CAP-desensitized compared to control atria. Cross-desensitization studies using CAP, NT and BK showed that the positive chronotropic and inotropic effects of CAP are slightly affected, whereas those evoked by BK are markedly reduced in NT-desensitized atria. On the other hand, the positive chronotropic and inotropic effects of CAP and NT were similar in BK-desensitized and control atria. The results were interpreted as an indication that NT, BK and CAP produce their excitatory effects in guinea pig atria by interacting with a common population of CAP-sensitive sensory nerve fibers (presumably substance P (SP)- and CGRP-containing nerve fibers). The absence of cross-desensitization between NT or BK and CAP, or between NT and BK, suggests that the activation and desensitization of atrial, CAP-sensitive sensory nerve fibers by the latter agents involve different receptors and/or mechanisms.