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In rat sinoatrial node, NAD(+) (10 μM) reduced the rate of spontaneous action potentials, duration of action potentials, and the velocity of slow diastolic depolarization, but the rate of action potential front propagation increases. In passed rabbit Purkinje fibers, NAD(+) (10 μM) reduced the duration of action potentials. Under conditions of spontaneous activity of Purkinje fibers, NAD(+) reduced the fi ring rate and the rate of slow diastolic depolarization. The effects of extracellular NAD(+) on bioelectric activity of the pacemaker (sinoatrial node) and conduction system of the heart (Purkinje fibers) are probably related to activation of P1 and P2 purinoceptors.

This study is aimed to the investigation of the nicotinamide adenine dinucleotide (NAD+) effects and mechanisms of action in a heart. NAD+ (mcM) induces multiphase alternation of contractile activity of isolated rat heart: short positive inotropic action is followed by a negative inotropic phase. NAD+ (1-100 mcM) induces decreasing of action potential duration (APD) in rat atrial myocardium (from 45 +/- 0.82 ms in control experiments to 39 +/- 1.05 (n = 8) and 32 +/- 2 (n = 8) during application of 10 and 100 mcM of NAD+, respectively). Significant APD increase (from 45 +/- 0.82 ms to 74 +/- 1.89 (n = 8) ms) was observed during washing out of NAD+ (100 mcM). ATP or adenosine was unable to increase APD both during application or washing out. NAD+ induced APD decrease was not suppressed by P1-antagonist theophylline. P1-purinoreceptor and metabolite independent direct action of NAD+ in rat heart is suggested. Activation of P2X or P2Y receptors, cyclic ADP-ribose accumulation in cardiomyocytes is proposed as a main mechanism of NAD(+)-induced effects in the heart.

In the mammalian myocardium, ACh, which is the main neurotransmitter of cardiac parasympathetic postganglionic fibres, can be released via both quantal (vesicular) and non-quantal (non-vesicular) mechanisms of secretion. Non-quantal release is continuous and independent of vagus activity and exocytosis of ACh-containing vesicles. During the incubation of myocardium in the presence of acetylcholinesterase (AChE) inhibitors, non-quantal ACh release leads to accumulation of ACh in the myocardium and cholinergic effects, which are proportional to the intensity of non-quantal secretion. The aim of the present study was to reveal whether non-quantal release of ACh can be modulated by another major cardioregulator, noradrenaline, or whether it represents uncontrolled leakage of ACh from cholinergic fibres. Cholinergic changes of electrical activity induced by the AChE inhibitor paraoxon (5 × 10(-6) M) in isolated rat right atrial preparations were determined by means of a standard microlectrode technique and used as a measure of the intensity of non-quantal release. Noradrenaline (10(-7) and 10(-6) M) substantially suppressed, but did not abolish, effects of paraoxon via stimulation of α-adrenoceptors, because all experiments were conducted in the presence of the β-blocker propranolol (5 × 10(-6) M). A blocker of ganglionic transmission, hexamethonium bromide (10(-4) M), failed to alter the inhibitory effect of noradrenaline, indicating that only non-quantal ACh release is suppressed by this neurotransmitter. The effects of noradrenaline could be reduced by the α2-antagonist yohimbine (10(-6) M). However, both the α1-agonist phenylephrine (10(-6) M) and the α2-agonist clonidine (10(-6) M) significantly inhibited the cholinergic effects of paraoxon, indicating the possible involvement of both α-adrenoceptor subtypes in mediation of the adrenergic inhibition of non-quantal ACh release. Thus, cardiac non-quantal ACh release can be negatively regulated by noradrenaline, providing another facet of sympathetic-parasympathetic interaction in the heart.

We studied the effect of selective activation of muscarinic M3receptors on electrical activity in the isolated preparation of rat ventricular myocardium as well as contractile activity of the left ventricle of Langendorff-perfused isolated heart. Application of muscarinic agonist pilocarpine (10(-5)M) against the background of selective blockade of subtype 2 muscarinic receptors with methoctramine (10(-7)M) markedly shortened the duration of action potentials in the isolated ventricular myocardium and reduced the amplitude and maximum rates of left-ventricular pressure rise and decay in the isolated heart paced at a fixed rate. All these effects were significantly suppressed by selective M3receptor blocker 4-DAMP (10(-8)M), which attested to the involvement of M3muscarinic receptors.

We studied the effects of carbon monoxide and sodium hydrosulfide, hydrogen sulfide donor, on contractile activity of the left ventricle in Langendorf-perfused isolated rat heart. Carbon monoxide 5×10(-5) M significantly accelerated sinus rhythm and left-ventricular pressure wave growth and decay. To the contrary, negative inotropic and chronotropic effects were observed at higher concentrations of carbon monoxide (10(-4), 3×10(-4) M). Sodium hydrosulfide (10(-4)-4×10(-4) M) decreased all the parameters of left-ventricular contractive activity and reduced contraction rate. Carbon monoxide and hydrogen sulfide, which together with nitrogen oxide are qualified as a new class of gaseous signal compounds, may substantially modulate pumping function of the heart.

The presence of M3 cholinoreceptors and their role in mediation of action potential waveform modulation were determined by immunolabeling of receptor proteins and standard microelectrode technique, respectively. The sinoatrial node (SAN), which was determined as a connexin 43 negative area within the intercaval region, the surrounding atrial tissue, and the working ventricular myocardium exhibited labeling of both M3 and M2 receptors. However, the density of M3 and M2 labeling was about twofold higher in the SAN compared to working myocardium. The stimulation of M3 receptors was obtained by application of nonselective M1 and M3 muscarinic agonist pilocarpine (10(-5) M) in the presence of selective M2 blocker methoctramine (10(-7) M). Stimulation of M3 receptors provoked marked shortening of action potential duration in atrial and ventricular working myocardium. In the SAN, M3 stimulation leads to a significant reduction of sinus rhythm rate accompanied with slowing of diastolic depolarization and increase of action potential upstroke velocity. All electrophysiological effects of selective M3 stimulation were suppressed by specific blocker of M3 receptors 4-DAMP (10(-8) M). We conclude that M3 cholinoreceptors are present in pacemaker and working myocardium of murine heart, where they mediate negative cholinergic effects: slowing of sinus rhythm and shortening of action potentials.

Standard microelectrode technique was used to study electrical activity of the isolated heart of the polychaete annelid, Arenicola marina. Typical pacemaker activity with slow diastolic depolarization was observed in all recordings. The average maximum diastolic potential (-58.4 +/- 3.2 mV), the average amplitude of the action potential (28.7 +/- 4.7 mV) and the average total duration of the action potential (2,434 +/- 430 ms) were determined. There has been no gradient of automaticity observed in our studies, which suggests that all regions of the Arenicola heart could possess pacemaker functions. Acetylcholine (ACh) produced a concentration dependent (5 x 10(-8)-5 x 10(-5) M) increase of the beating rate via increase in the rate of the diastolic depolarization. ACh (5 x 10(-5) M) increased beating rate by 2.5-fold compared to the control rate. A stronger action of ACh resulted in depolarization, block of action potential generation and contracture of the heart. The non-hydrolysable ACh analog carbacholine (10(-8)-10(-6) M) produced similar effects. All effects of ACh and carbacholine were abolished by 5 x 10(-6) M atropine. D-Tubocurarine (5 x 10(-5) M) did not significantly alter effects of ACh or carbacholine. Epinephrine (10(-8)-10(-6) M) caused the slowing of pacemaker activity and marked decrease of action potential duration. 10(-6) M epinephrine produced complete cardiac arrest. The effects of epinephrine were not significantly altered by the beta-blocker propranolol (5 x 10(-6) M). The beta-agonist isoproterenol (10(-7)-10(-5) M) and the alpha-agonist xylometazoline (10(-6)-10(-5) M) did not produce significant effects. Thus, cholinergic effects in the Arenicola heart are likely to be mediated via muscarinic receptors, while the nature of adrenergic effects needs further investigation.

Changes of the activation sequence in the rabbit sinoatrial node under the influence of low temperature and I(f) selective blocker ivabradine have been studied using the optical mapping technique. Both factors caused a shift of the pacemaker within the sinoatrial node region. These results are compared with the data obtained recently in the investigation of pacemaker shift under the influence of cholinergic and adrenergic factors. Possible mechanisms of the pacemaker shift are discussed. The suppression of electric activity in the central part of the sinoatrial node during the action of acetylcholine, which is called cholinergic inexcitability, may be considered as one of the mechanisms of the pacemaker shift. It is shown that the main cause of cholinergic inexcitability is the activation of potassium acetylcholine-dependent current I(KACh).

Acetylcholinesterase (AChE) inhibitors provoke typical cholinergic effects in the isolated right atrium of the rat due to the accumulation of acetylcholine (ACh). Our study was designed to show that in the absence of vagal impulse activity, ACh is released from the parasympathetic nerve fibres by means of non-quantal secretion. The conventional microelectrode technique was used to study changes in action potential (AP) configuration in the right atrium preparation of rats during application of AChE inhibitors. Staining with the lipophilic fluorescent dye FM1-43 was used to demonstrate the presence of endocytosis in cholinergic endings. The AChE inhibitors armin (10(7)-10(5)m) and neostigmine (10(7) to 5 x 10(6)m) caused a reduction of AP duration and prolonged the cycle length. These effects were abolished by atropine and were therefore mediated by ACh accumulated in the myocardium during AChE inhibition. Putative block of impulse activity of the postganglionic neurons by tetrodotoxin (5 x 10(7)m) and blockade of ganglionic transmission by hexomethonium (2 x 10(4)m), as well as blockade of all forms of quantal release with Clostridium botulinum type A toxin (50 U ml(1)), did not alter the effects of armin. Experiments with FM1-43 dye confirmed the effective block of exocytosis by botulinum toxin. Selective inhibition of the choline uptake system using hemicholinium III (10(5)m), which blocks non-quantal release at the neuromuscular junction, suppressed the effects of AChE inhibitors. Thus, accumulation of ACh is likely to be caused by non-quantal release from cholinergic terminals. We propose that non-quantal release of ACh, shown previously at the neuromuscular junction, is present in cholinergic postganglionic fibres of the rat heart in addition to quantal release.

Cholinergic changes of electric activity were studied in isolated atrium preparations from fishes (cod and carp), amphibians (frog) and reptilians (lizard) using the microelectrode technique and high-resolution optical mapping. Perfusion of isolated atrium with acetylcholine (10(-6)-5.10(-5) M) caused gradual suppression of action potential generation and, eventually, completely blocked the excitation in a part of the preparation. Other regions of atrium, situated close to the sinoatrial and atrioventricular junctions, remained excitable. Such cholinergic suppression of electric activity was observed in the atrial myocardium of frog and in both fish species, but not in reptilians. Ba(2+) (10(-4) M), which blocks the acetylcholine-dependent potassium current (I(KACh)), prevented cholinergic reduction of action potential amplitude. In several preparations of frog atrium, cholinergic suppression of excitation coincided with episodes of atrial fibrillation. We conclude that the phenomenon of cholinergic suppression of electric activity is typical for atria of fishes and amphibians. It is likely to be caused by I(KACh) activation and may be important for initiation of atrial arrhythmias., (2009 Elsevier Inc. All rights reserved.)

Actual data concerning mechanisms of automaticity in sinoatrial node, which acts as a primary pacemaker in mammalian heart, is reviewed. Studies dealing with ionic currents, maintaining automatic generation of excitation in the sinoatrial cells, and possible role of intracellular calcium turnover are discussed. Special attention is given to the differences between the central and peripheral parts of sinoatrial node, phenomenon of intranodal pacemaker shift resulting from that differences and possible role of pacemaker shift in the modulation of the sinus rhythm. Mechanisms of sinus rhythm regulation under the action of acetylcholine and noradrenalin are also discussed in detail.

Aims: Changes in the rabbit sinoatrial node (SAN) activation sequence with the cholinergic and adrenergic factors were studied. The correlation between the sinus rhythm rate and the leading pacemaker site shift was determined. The hypothesis concerning the cholinergic suppression of nodal cell excitability as one of the mechanisms associated with pacemaker shift was tested., Methods: A high-resolution optical mapping technique was used to register beat-to-beat changes in the SAN activation pattern under the influence of the cholinergic and adrenergic factors., Results: Acetylcholine (10 microm) and strong intramural parasympathetic nerve stimulation caused a pacemaker shift as well as rhythmic slowing and the formation of an inexcitable region in the central part of SAN. In this region the generation of action potentials was suppressed. The slowing of the sinus rhythm (which exceeded 12.8 +/- 3.1% of the rhythm control rate) always accompanied the pacemaker shift. Isoproterenol (10, 100 nm, 1 microm) and sympathetic postganglionic nerve stimulation also evoked a pacemaker shift but without formation of an inexcitable zone. The acceleration of the sinus rhythm, which exceeded 10.5 +/- 1.3% of the control rate of the rhythm, always accompanied the shift., Conclusions: Both cholinergic and adrenergic factors cause pacemaker shifts in the rabbit SAN. While modest changes in the sinus rhythm do not coincide with the pacemaker shift, greater changes always accompany the shift and may be caused by it, according to one hypothesis. The formation of an inexcitable zone at the place where the leading pacemaker is situated is one of the mechanisms associated with pacemaker shift.

Physiological effects of kiotorphyn (KT) and neokiotorphyn (NKT) has been reviewed. These peptides were isolated from the brain of hibernating ground squirrel Citellus undulates (CU) and assumed to be endogenous regulators of hibernation NKT does not alter the process of entering to hibernation, but induces arousal from the sleeping. KT administration in squirrel delays the rise of heart rate and body temperature associeated with awakening provoked in the middle but not at the final stage of hibernation bout. Neither KT nor NKT induce physiological effects in rats and mice under normal environmental conditions. In mice with induced hypothermia (17-18 degrees C) NKT accelerates restoration of body temperature, whereas KT induces progressive hypothermia. In addition, NKT modulates cardiotropic effects of nucleotides found in physiologically active fractions of hibernating squirrel tissues. NKT as well as KT modify sensitivity of cardiovascular system to adrenaline and AMP in rats one day after hypothermia.

Changes of electric activity induced by acetylcholine were studied in atrial myocardium of fishes (cod and carp) and reptilians (lizard and grass-snake). Standart microelectrode technique and novel method of optical mapping were used in the study. Acetylcholine (1-50 microM) provoked decrease of the action potential amplitude down to full inhibition of electrical activity in wide regions of atrium of cod and carp. We define this phenomenon as cholinergic inexcitability. In other regions excitation persisted even during action of 500 microM acetylcholine. In atria of lizard and grass-snake acetylcholine caused shortening of action potential without changes in it's amplitude. Local cholinergic inexcitability, shown in the atrial myocardium of fishes, is quite similar to the phenomenon, that was described earlier in the atria of frogs. It presents the heart of fish as an interesting model for study of mechanisms of cholinergic atrial arrhythmias initiation.

We used high resolution optical mapping for the study of activation sequence in a rabbit sinoatrial node in normal state and during artificial stimulation as well as changes of excitation chronotopography under cholinergic influences. In the norm excitation originates in central region of sinoatrial node. During acetylcholine action (10, 50, 100 M) and activation of intramural parasympathetic nerves suppression of electrical activity in the zone of initial pacemaker localization occurs. Suppression of activation is associated with pacemaker migration within sinoatrial node and lowering of the rate of generation of excitation. Termination of cholinergic influences has been followed by restoration of initial chronotopography of excitation. During artificial stimulation of sinoatrial node nonexitable (refractory) area has been formed in the region of initial localization of the natural pacemaker.

Experimental facts, proving the presence of M3-cholinoreceptros in parasympathetic regulation of heart, are reviewed. Results of molecular biological and biochemical studies, showing existence of M3-cholinoreceptors in myocardium, are discussed. Physiological data, that demonstrates changes of action potential configuration due to M3-receptors activation, specific potassium current, linked with M3-receptors stimulation, and other evidence of functional importance of M3-receptros in myocardium, is analyzed. Studies dealing with cardioprotective role of M3-receptors are reviewed with special attention. The review widens our knowledge about molecular mechanisms of parasympathetic heart regulation.

We used high resolution optical mapping for the study of changes of activation sequence of a rabbit sinoatrial node induced by adrenergic stimulation during natural and paced rhythm. Activation of adrenoreceptors with isoproterenol (10, 100 nanoM, 1 microM) as well as stimulation of intramural postganglionic sympathetic nerves caused pacemaker migration within sinoatrial node and increase of the rate of generation of excitation. Pacemaker migration in cases of pronounced acceleration of rhythm could proceed in two stages. Termination of adrenergic influences has been followed by restoration of initial chronotopography of excitation of sinoatrial node.

The aim of work was to investigate effects of adenosine, AMP, GMP and ADP-ribose on bioelectric activity of bat heart. Purine nucleotides decreased action potential duration at level of 90% (APD90) repolarization in bat ventricular myocardium. When preparation of right ventricle was paced with frequency of 6 Hz, APD50 and APD90 were 7 +/- 2 ms and 60 +/- 5 ms, respectively. Adenosine decreased APD90 by 50 +/- 10% (n=6), AMP - by 30 +/- 10% (n=6), GMP - by 38 +/- 5% (n=6), ADP-ribose - by 24 +/- 6% (n=6) (concentration of all compounds - 10 ). Effects of purine nucleotides and nucleoside in bat ventricular myocardium exceed effects of same compounds in rat and ground squirrel ventricular myocardium. Inhibitory effects of purine nucleotides and adenosine in bat heart could be mediated by A1 adenosine receptors.

The aim of work was to investigate effects of adenosine, AMP and ADP-ribose (1x10(-5)) on bioelectric activity of atrium and papillary muscle of nonhibernating (rat) and hibernating (Yakutian ground squirrel) animals. Action potential (AP) was registered with use of standard microelectrode technique. AP duration (APD) at level of 90% repolarisation in rat atrium in control experiments was 30+/-5 ms, APD at level of 50% repolarisation was 12+/-2 ms. APD at level of 90% repolarisation in rat papillary muscle was 56+/-7 ms, at level of 50% repolarisation was 18+/-2 ms. APD at level of 90% repolarisation in ground squirrel atrium was 77+/-6, APD at level of 50% repolarisation was 38+/-6 ms. APD at level of 90% repolarisation in ground squirrel papillary muscle was 105+/-9 ms, APD at level of 50% repolarisation was 42+/-8 ms. Purine nucleotides and nucleoside, that were tested in work, except ADP-ribose, act as inhibitory factors and decrease APD both in rat and hibernating ground squirrel heart. ADP-ribose decreases APD in papillary muscle of hibernator but did not in its atrium. In ground squirrel atrium AMP and adenosine decrease APD at level of 50% repolarisation by 10+/-3% and 18+/-3% respectively. AMP and adenosine decrease APD at level of 90% repolarisation by 9+/-2% and 11+/-2% respectively. In ground squirrel papillary muscle ADP-ribose, AMP and adenosine decrease APD at level of 50% repolarisation by 26+/-8%, 23+/-8% and 26+/-7%. ADP-ribose, AMP and adenosine decrease APD at level of 90% repolarisation by 12+/-3%, 10+/-3%, 13+/-3%. Thus, decrease of APD in ground squirrel papillary muscle at level of 90% repolarisation during nucleotides and adenosine action was 2-2.5 fold less, than the rat.

The excitation of the sinoatrial unit from heart auricles of the frog has been studied. Potentials were recorded by means of microelectrodes inserted to pacemaker of the sinoatrial unit. It has been established that auricles can impart the rhythm to the sinoatrial unit due to electric and electromechanical influence, and electromechanical influence is of greater significance. Specific transitions accompanying the establishment of the stationary rhythm have been studied. A mathematical model of transitions of the establishment of the rhythm of the sinoatrial unit, which is based on diophant methods is offered. The catculations performed by means of the mathematical model coincide well with results of experimental studies. The stabilizing role of auricles in the formation of the rhythm of the sinoatrial unit has been established.

We studied acetylcholine induced changes of action potentials (AP) in ventricles and atria of bat, hedgehog, yakut ground squirrel, and laboratory mice. In atria of all these species of mammals acetylcholine caused pronounced dose dependent shortening of AP. However sensitivity of ventricles of studied animals to acetylcholine was different. Acetylcholine induced AP shortening was 29.4, 42.1, 6.6 and 11% in ventricles of bat, hedgehog, laboratory mice and ground squirrel, respectively. Ventricles of none of previously studied mammals had sensitivity to acetylcholine comparable by magnitude to acetylcholine sensitivity which we found in hedgehog and bat. It is suggested that these animals have unique nervous regulation of cardiac action, principally different from that of other mammals, which do not require parasympathetic regulation of ventricles.

Functions and possible mechanisms of action of adenosine diphosphate ribose (ADP-ribose) and its cyclic analogue--cycle-ADP-ribose (cADP-ribose)-- substances pretend to role of novel second messengers are reviewed. Possible mechanisms of ADP-ribose and cADP-ribose synthesis regulation and metabolism are analyzed. Prospective mechanisms of ADP-ribose and cADP-ribose transportation into the cell are described. ADP-ribose and cADP-ribose are considered as endogenous regulators of ionic balance (in particular regulators of calcium balance) in different tissues. Effects of ADP-ribose on isolated heart of frog and warm-blooded animal (rat), as well as on rat heart in vivo are discussed. The role of ionic channels and receptors, gated by ADP- and cADP-ribose in regulation of cardiomyocyte ionic balance in normal and pathological condition are analyzed. The possible role of purine receptors in ADP-ribose and cADP-ribose effects on heart are discussed.

Cardiotropic effects of endogenous regulator of intracellular ionic balance adenosine diphosphate ribose (ADP-ribose) was studied on isolated rat heart using standard retrograde perfusion and on rat heart in vivo by means of implanted arterial catheters. Parameters registered were heart rate, pressure developed in left ventricle of isolated heart and blood pressure of rats in vivo. It was shown that ADP-ribose exerted cardiotropic action on isolated rat heart and that its effects did not depend on effects of its metabolite AMP. In concentrations 10-70 microM ADP-ribose caused negative dose dependent chronotropic effect. Amplitude of contraction of isolated heart changed equivocally. In concentration 70 microM ADP-ribose in 50% of cases produced biphasic action: negative inotropic effect was preceded by positive inotropic effect. In other cases this concentration exerted negative inotropic effect. In concentration 10 microM only negative inotropic effect was observed. Contrary to ADP-ribose its metabolite AMP exerted only monophasic positive inotropic effect on isolated heart. The preparations studied differed in dynamics of cardiotropic action. In experiments in vivo ADP-ribose and AMP caused monophasic negative chronotropic and monophasic positive inotropic effects.

Direct effects of cardiotropic preparations on the hearts isolated from Wistar rats were examined. Deenergization of cardiomyocytes was modeled under conditions of hypoxic perfusion. Recovery of cardiac function during reperfusion was assessed by changes in the heart rate and contraction amplitude.

Heart rate (HR) of ground squirrel C. undulatus was studied in dependence of season, level of activity, physiological state and air temperature (T). In summer HR varies from 110-130 beat/min in sleep up to 420 beat/min at flight from danger. During winter hibernation HP was minimal (3-5 beat/min) at T 1-4 degrees C, the increase in T induced the growth of HP in correspondence to the Arrenius van't Hoff law. The temperature of the body in hibernation exceeded T on 1.5-3 degrees C. The time of getting off the hibernation increased with the decrease in T (6-7 hours at -1 degree C and 2.5-3 hours at +18 degrees C). At phase of increased thermogenesis during arousal heart temperature exceeded rectal one on 10-12 degrees C and heart rate run up to 360-420 beat/min i.e. 2-3 time higher than in active state. The decrease in T stimulated the increase in HP up to 3.8 in winter and 5.3 beat/min/degree C in summer. The highest values of Q10 for HP were revealed at the beginning of hibernation (15-20) and at the beginning of arousal (6-7), in other periods Q10 was similar to the normal biological values (2-2.5). Thus, at the beginning of transitional periods changes of HP were determined mainly by endogenic mechanisms that inhibited myocardium at the beginning of hibernation and activated in arousal. Some mechanisms of coordination between activities of heart and other systems of organism are considered. The features of hit exchange providing the hibernation in wide range of T are discussed.

In the frog isolated heart, cyclic perfusion of ADP-ribose induced a dose-dependent decrease in the heart rate and the contraction force, a decrease in the AP duration as well as in the rate of rise in the sinus node. It also shortened the atrial AP and exerted no significant effect upon multicellular ventricular preparations. In conditions of systemic administration in unanesthetised frogs, the ADP-ribose induced a reversible increase in the heart rate due, probably, to a sympathetic effect.

It was supposed that the transitional period from euthermia to hibernation rather than the period of winter hibernation is optimal for the secretion of biologically active substances in rodent tissues. This hypothesis was tested in experiment. The maximal O2 consumption suppression was noted in mice after I. P. Injection of blood plasma from suslik Citellus undulatus at the beginning of its entering into hibernation. Low-molecular peptide fraction appearance in C. undulatus blood before hibernation was revealed by electrophoresis in gel and autoradiography. But its traces disappear by the end of bout. The endogenic inhibitory factor's maximal influence in early bout was shown by experiments with TRH and neoklotorphin (NKT). I.P. injections of these substances to C. undulatus were ineffective in the beginning of entering into hibernation, but clearly caused awakening after hibernation, has started. KT, being a fragment of NKT, inhibited suslik's heart rate when the awakening was provoked in the middle, but not in the end of bout. Possible transformation of biologically active substances in winter-hibernating rodents tissues is discussed. The KT "inhibitor" and NKT "stimulator" are supposed to pertain to the special type of regulatory peptides, that manage the hibernation cycle. One of the ways of inactivation of endogenic "hibernation trigger" is its removal from the organism with urine. The urine, taken from suslik immediately after hibernation proved to produce the greatest hypothermic effect on mice, I.P. injected with it.

More than 20 peptides have been isolated and sequenced from the brain of hibernating ground squirrel Citellus undulatus. Some of the isolated peptides were chemically synthesized and investigated for the spectrum of biological activity. One of the isolated peptides, neokyotorphin (Thr-Ser-Lys-Tyr-Arg), earlier known as a weak analgetic, is found to have a cardiotropic and thermoregulatory activity. Neokyotorphin activates in vitro voltage-dependent calcium and blocks ATP-dependent potassium currents in the frog atrial fibres. Intraperitoneal injection of this peptide in hibernating ground squirrels speeds up the arousal of animals increasing sharply the heart rate and oxygen consumption. Intraperitoneal and intranasal administrations of neokyotorphin in rats raises body temperature in thermoneutral conditions (26-28 degrees C) exerting no effects at low (4-6 degrees C) and high (31-32 degrees C) environmental temperatures. Another isolated peptide, Asp-Tyr, blocks the inward voltage-dependent calcium current in the frog atrial fibres and slightly increases the outward potassium current. Sulfated analogue of this dipeptide (aspartyl-O-sulfate-tyrosine) more effectively blocks the inward voltage-dependent calcium current in the frog atrial fibres and have no effects on the outward potassium current.

Decrease of ambient temperature (Ta) leads to the increase of the heart rate (HR) in active ground squirrels C. undulatus by 5.3/min/1 degree C in summer and by 3.8/min/1 degree C in winter. In a hibernation state, the dependence of the HR on Ta was in a good agreement with equation HT = 2.53.exp.(0.1.Ta). On entering into hibernation and on arousal, the HR change outruns the corresponding body temperature (Tb) change by 1.5-2 hours. A maximum HR level (up to 400/min and more) was registered on arousal when Tb reached 17-20 degrees C. A minimal HR level (4-5/min) was observed during hibernation at Ta 2-5 degrees C. The maximum Ta, at witch C. undulatus was hibernating, reached 23-24 degrees C, the HR being 23-25/min.

The effect of intraabdominal injection of the blood serum from ground squirrels Citellus undulatus obtained at various stages of the baut has been studied on the level of oxygen consumption in albino mice. The strongest hypometabolic effect exhibited the blood plasma from animals at the beginning of the baut. The onset of baut was used in further experiments on C. undulatus for investigation of activating effects of neokiotorphin (Thr-Ser-Lys-Tyr-Arg) which was earlier found in the brain of the ground squirrel. It was shown that intraabdominal injection of this peptide to hibernating animals facilitates their arousal stimulating both the cardiac activity and respiration. The strongest effect was noted with neokiotorphin of [D-Ser] 2-modification which is more stable to proteolysis.

In experiments on active ground squirrels, it has been shown that the effect of intraperitoneal injection of serotonin on the heart rate depends on the dose and the environmental temperature. At relatively low doses and high environmental temperature, serotonin increases the heart rate and body temperature; on the contrary, with the increase in the dosage and the decrease in the environmental temperature the heart rate and body temperature of animals decrease. It is suggested that this relationship serves as one of the factors which enhance the transition of animals from one level of activity to another during the beginning and the end of hibernation.

The main physiologic hypotheses in establishing a common rhythm of the heart sino-atrial node (SAN) have been considered. A mathematical model of SAN which takes into account the interaction between pace-makers in pairs has been proposed. A quantitative description of common rhythm establishing taking into account interaction in pairs has been obtained, the expressions for the period of common rhythm, a delay and the rate of stimulation distribution have been found. A comparison of calculated and experimental data is given.

This study was designed to examine the changes of the isolated frog's heart rate as a function of the time in the two time intervals: 20-95 min and 95-170 min after hearts were prepared. The heart rate decreased in these intervals quite linear but average slope between 20-75 min was significantly steeper than in the time interval 95-170 min. It was shown that the difference in the heart rate response induced by the increasing temperature in intact animals and isolated hearts partly might be explored by this decrease in the isolated frog's heart rate with time.

From tissues of hibernating and active long-tailed ground squirrels and from the brain of cold-adapted Yakut horses, low molecular peptide fractions were obtained which, after injection to albino mice, decreased oxygen consumption and rectal temperature in them. The same fractions exhibited negative chrono- and inotropic effects on isolated hearts of ectothermic and endothermic animals. Fractions from the brain of ground squirrels and the brain of horse exhibited similar pattern of the activity. The activity of fractions was subjected to seasonal changes and depended on the degree of their purification. Provisional intracellular microelectrophysiological analysis of the effect of these fractions on the frequency and strength of contractions in isolated heart was made.

Acceleration of the heart rate can be elicited by stimulation of the extracardiac paraysmpathetic pathways (previous investigations). In this study, changes of the pacemaker APs were continually recorded with microelectrode in the sinus node cell. The data showed acceleration of the slope of slow diastolic depolarization (distolic prepotential) of the pace-maker AP during the parasympathetic positive chronotrct. The parasympathetic acceleration of the slow diastolic depolarization and of the heart rate was shown to be cholinergic by its nature.

Variations in the sinus node rate of the frog heart isolated together with the atria were studied during suprathreshold atrial stimulation at a frequency exceeding the intrinsic sinus node rate. If the stimulation frequency surpassed the intrinsic sinus node rate by 20%, the sinus node rate became equal to the pacing rate. Provided the pacing rate was increased, the sinus node rhythm became irregular. If the pacing rate exceeded the sinus node rate more than two-fold, the latter became half the pacing rate. Variations in the sinus node rate during suprathreshold atrial stimulation were also observed after blockade of possible retrograde conduction by application of the necrotic tissue on the sinoatrial border. The results allow the conclusion that variations in the sinus node rate seen during high-frequency suprathreshold atrial stimulation are not connected with retrograde conduction.

The cardiac pacemaker and atria were removed from the frog heart and placed into a chamber with Ringer solution. Atrial pacing was performed at a suprathreshold milliamperage. The mechanism of changes in the cardiac pacemaker during atrial pacing was studied both under the conditions of the blockade of the intracardiac nervous system and elimination of the bioelectrical and mechanical effects of the atria on the cardiac pacemaker. It is inferred that the change in the rate of pacemaker stimulation during atrial pacing is largely determined by the mechanical activity of the atria.

The automatic activity of the venous sinus was studied in respect to the atrial bioelectrical activity in frogs. Each of the two parts of the divided venous sinus had different rhythms. Extra-cellular recording revealed that the APs of both parts of the sinus were closely synchronized if the parts were connected with the atrium. The data obtained suggests that this synchronization was due to atrial bioelectrical activity.

Two pacemakers localized in sinoatrial and atrioventricular valves were revealed in the tench heart. In their absence, the ventricular and sometimes the atrial myocardium are capable of initiating the automaticity. Pacemaker's cells of the sinoatrial valve are surrounded by specific area similar to sinoauricular ring in amphibia or sino-caval region in mammals. Characteristics of the action potential in different parts of the tench heart are presented.