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.     

Photosensitive neurogenic heart of the isopod crustacean Ligia exotica

The heart of animals is regulated through the central nervous system in response to external sensory stimuli. We found, however, that the adult neurogenic heart of the isopod crustacean Ligia exotica has photosensitivity. The beat frequency of the isolated heart decreased in response to a light stimulus. Magnitude of the response was stimulus intensity dependent and the heartbeat frequency decreased to less than 80% of the dark value during illumination of the white light with an intensity of 6.0 mW cm-2. The spectral sensitivity curve of the heart photoresponse peaked at a wavelength around 520 nm. In response to 530 nm monochromatic light, the relationship between light intensity and response magnitude was linear and the threshold intensity was 7.26 x 1012 quanta cm-2 s-1. Bursting activity of the cardiac ganglion, which is located in the heart and acts as the cardiac pacemaker deceased in frequency in response to illumination by white light. This fact suggests that the heart photoresponse of L. exotica results from the photosensitivity of the cardiac ganglion neurons. The photoresponse of the heart therefore contributes to regulation of cardiac output in addition to other regulatory systems.     

Dual effects of dopamine on the adult heart of the isopod crustacean Ligia exotica

In the adult heart of the isopod crustacean Ligia exotica, the cardiac ganglion acts as the primary pacemaker with the myocardium having a latent pacemaker property. We show several lines of evidence that dopamine modulates the heartbeat of adult L. exotica affecting both pacemaker sites in the heart. Dopamine caused positive chronotropic (frequency increase) and inotropic (amplitude increase) effects on the heartbeat in a concentration dependent manner. The time courses of these effects were considerably different and the inotropic effect appeared later and lasted longer than the chronotropic effect. Dopamine rapidly increased the frequency of the bursting activity in the cardiac ganglion neurons and each impulse burst of the cardiac ganglion was always followed by a heartbeat. Moreover, dopamine slowly increased the amplitude and duration of the action potential plateau (plateau potential) of the myocardium. When the myocardial pacemaker activity was induced by application of tetrodotoxin, which suppresses cardiac ganglion activity, dopamine slowly increased the amplitude and duration of the myocardial plateau potential while decreasing its frequency. These results suggest that dopamine modulates the heartbeat in adult L. exotica producing a dual effect on the two pacemaker sites in the heart, the cardiac ganglion and myocardium.     

Cardioacceleratory Neurons of the Isopod Crustacean, Ligia exotica: Visualization of Peripheral Projection onto the Heart Muscle

Innervation of the heart muscle by the cardioacceleratory neurons was morphologically and electrophysiologically examined in the isopod crustacean, Ligia exotica. Intracellular injection of neurobiotin into the first and second cardioacceleratory neurons (CA1 and CA2) revealed their peripheral axonal projections. Inside the heart, the CA1 and CA2 axons ran along the trunk of the cardiac ganglion. Finely arborized branches with many varicosities arose from the axon and projected over the heart muscle. Stimulation of either the CA1 or CA2 axon caused an overall depolarization in the muscle of a quiescent heart. The amplitude of the depolarization increased with increasing stimulus frequency. During stimulation, the membrane resistance of the heart muscle decreased. In a beating heart, the cardioacceleratory nerve stimulation caused multiple effects on the heart muscle activity and the heartbeat. The results suggest that the cardioacceleratory neurons of Ligia exotica regulate the amplitude of the heartbeat (inotropic effect) and the heart tonus (tonotropic effect) via the synaptic contacts on the heart muscle, while the heartbeat frequency (chronotropic effect) is regulated via the synapses on the cardiac ganglion neurons.     

Acceleratory nervous regulation of juvenile myogenic hearts in the isopod crustacean Ligia exotica

We examined regulation of the myogenic heart by two identified cardioacceleratory neurons (CA1, CA2) in early juveniles of the isopod Ligia exotica. Repetitive stimulation of either the CA1 or CA2 axon increased the frequency and plateau amplitude of the action potential and decreased the maximum hyperpolarization of the cardiac muscle. These effects were larger with increasing stimulus frequency. The rate of increase in the frequency caused by CA1 stimulation was significantly larger than that by CA2. No impulse activity of the cardiac ganglion was induced by acceleratory nerve stimulation. The frequency of the muscle activity was decreased by injection of a hyperpolarizing current into the muscle during stimulation of the acceleratory nerve. In a quiescent heart, acceleratory nerve stimulation caused an overall depolarization in the muscle membrane and the amplitude of the depolarization induced by CA1 stimulation was significantly larger than that by CA2. These results suggest that CA1 and CA2 neurons regulate the myogenic heart affecting directly the cardiac muscle; the CA1 neuron produces more potent effects than does the CA2 neuron.     

Cardiac pacemaker mechanisms in the ostracod crustacean Vargula hilgendorfii

The heart of the ostracod crustacean Vargula hilgendorfii has a single intrinsic neuron that morphologically appears to innervate the myocardium. We, therefore, examined the heart activity electrophysiologically to determine whether the heartbeat is neurogenic. Each heartbeat is associated with a myocardial action potential composed of a spike potential followed by a plateau potential. The frequency of the action potential is not stable but changes successively over a wide range. The action potential is not preceded by a pacemaker potential and has an inflection in its rising phase. The myocardial cells couple electrically and fire almost simultaneously. The frequency of the action potential was unchanged by injection of depolarizing or hyperpolarizing current into the myocardium. However, slow oscillatory potentials appeared during the depolarization and its frequency was higher with increasing current intensity. Application of 1-microM tetrodotoxin (TTX) depolarized the myocardial membrane and completely prevented the action potential. During this depolarization, slow oscillatory potentials often appeared spontaneously. These results suggest that, although the myocardium has a property of conditional oscillator, the heartbeat is driven by the single cell cardiac ganglion that has both pacemaker and motor functions.     

TEMPERATURE AND FREQUENCY OF CARDIAC CONTRACTIONS IN EMBRYOS OF LIMULUS

Temperature characteristics for frequency of myogenic heart beat in Limulus embryos, before the onset of nervous control of the heart, were found to be 11,500; 16,400; 20,000; 25,500. The two first values are the best established. The different values pertain to the hearts of different individuals outwardly similar, and to the hearts of single embryos in different parts of the temperature range. These values differ from that known in connection with the control of the heart beat through the cardiac ganglion. The occurrence of critical temperatures, also, is not the same in all embryos. These facts are employed in a discussion of temperature relations in pulsating explants of chick myocardium.     

Ontogeny of neurohormonal regulation of the cardiovascular system in the crayfish Procambarus clarkii

Adult crayfish have a neurogenic heart which is modulated via inputs from the central nervous system and neurohormones, which act on the cardiac ganglion or directly on the myocardium. This study investigates the ontogeny of cardiac regulation by exploring the temporal sequence of cardiac sensitivity to injections of cardioactive neurohormones (proctolin, serotonin and octopamine) and the neurotransmitter gamma-aminobutyric acid. The cardiac response (delta in heart rate, stroke volume, or in cardiac output) to each neurohormone at each developmental stage was assessed. The observed response elicited by each cardioactive drug was stage dependent and changed as the animals progressed from embryonic through larval and juvenile periods. During early developmental stages, octopamine, serotonin, and proctolin (10(-9)-10(-3) M) did not result in a modulation of stroke volume, yet in later developmental stages they caused significant increases in stroke volume, at comparable concentrations. Early developmental stages are capable of regulating cardiac function, however, the mechanisms appear to be quite different from those used by adults. Evidence is also provided to support the hypothesis that cardiac function is initiated prior to the establishment of an adult-like regulatory system.     

Contributions on cardiac physiology in Diplopoda (Myriapoda)

Investigations into cardiac physiology in Myriapoda are rare, but heart beat generation is not considered to be uniform throughout this taxon. Although cardiac automatism in Chilopoda is neurogenic, superimposed onto a myogenic automatism, the present study reveals, on the basis of electrophysiological experiments including electrocardiograms and the first intracellular recordings from dorsal vessel muscle cells of Archispirostreptus gigas Peters, 1855 (Diplopoda: Spirostreptida, Spirostreptidae), that heartbeat generation in Diplopoda is clearly myogenic. Experiments with tetrodotoxin confirm this result, and also show that proctolin, acetylcholine and octopamine have no effect. The results are discussed from the perspective of comparative cardiac physiology in arthropods.     

A quantitative comparison of the effects of intracellular calcium injection and light adaptation on the photoresponse of Limulus ventral photoreceptors

Calcium ions were iontophoretically injected into ventral photoreceptors of Limulus by passing current between two intracellular pipettes. Changes in sensitivity and photoresponse time course were measured for both light adaptation and Ca++ injection. We found for some photoreceptors that there was no significant difference in the photoresponse time course for desensitization produced by light adaptation or by Ca++ injection. In other photoreceptors, the time delay of photoresponse for Ca++ injection was slightly longer than for light adaptation. The variability of threshold response amplitude and time delay decreases when the photoreceptor is desensitized by either light adaptation or Ca++ injection. The peak amplitude versus log stimulus intensity relationships for controls, light adaptation, and Ca++ injection all could be described very closely by a single template curve shifted along the log intensity axis. A 40- to 50-fold change in sensitivity is associated with a 2-fold change in photoresponse time delay for both light adaptation and Ca++ injection.     

Inhibitory Synapses on Pacemaker Neurons in the Heart Ganglion of a Stomatopod, Squilla oratoria

The pacemaker neurons of the Squilla heart ganglion are innervated from the CNS through three pairs of extrinsic nerves. One of them, the α-nerve, is inhibitory to the heart beat. The effect of α-nerve stimulation on the pacemaker potential was examined with intracellular electrodes. Without extrinsic nerve stimulation the membrane potential of the pacemaker cell fluctuated spontaneously. On application of a tetanic train of stimuli to the α-nerve the membrane potential was shifted and fixed to a steady level, which with K₂SO₄-filled electrodes was near the peak of hyperpolarization after a spontaneous burst, but was less negative with KCl-filled electrodes. The shift of the membrane potential was due to the summated IPSP's. By changing the level of the membrane potential with injection of the polarizing current the IPSP could be reversed in sign, and the size of the IPSP was linearly correlated with the membrane potential level. During inhibition the membrane conductance increased. The increase depended on divalent cation concentrations in the outside medium. In Ca-rich saline the IPSP was greatly enhanced. In Mg-rich saline it was suppressed. The amplitude of antidromic spikes was reduced during inhibition especially when the spike frequency was high.     

Photoresponse and Learning Behavior of Ascidian Larvae, a Primitive Chordate, to Repeated Stimuli of Step-Up and Step-Down of Light

Ascidians are lower chordates and their simple tadpole-like larvae share a basic body plan with vertebrates. Newly hatched larvae show no response to a stimulus of light. 4 h after hatching, the larvae were induced to swim upon a step-down of light and stop swimming upon a step-up of light. At weaker intensity of light, the larvae show the same response to a stimulus after presentation of repeated stimuli. When intensity of actinic light was increased, the larvae show sensitization and habituation of the swimming response to a stimulus after repeated stimuli of step-down and step-up of the light. Between 2 h 20 min and 3 h 40 min after hatching the larvae did not show any response to the first stimulus, but after several repeatedstimuli they show swimming response to a step-down of light. A repeated series of stimulus cause sensitization. Between 4 h and 7 h after hatching, the larvae show photoresponse to the first stimulus, but after several repetition of the stimuli, the larvae could not stop swimming to a stimulus of a step-up of the actinic light. A repeated series of stimulus cause greaterhabituation. Both sensitization and habituation depend upon intensity ofactinic light.     

Aminergic modulation of the myogenic heart in the branchiopod crustacean Triops longicaudatus

Although crustaceans typically have a neurogenic heart, the primitive crustacean Triops longicaudatus has a myogenic heart with the heartbeat arising from the endogenous rhythmic activity of the myocardium. In the present investigation, the effects of six biogenic amines, epinephrine, norepinephrine, dopamine, octopamine, serotonin and histamine, on the myogenic heart of T. longicaudatus were examined. Epinephrine, norepinephrine, dopamine and octopamine accelerated the heartbeat, increasing both the frequency and amplitude of the action potential of the myocardium in a concentration dependent manner. The ability of epinephrine and norepinephrine to produce the acceleratory effects was more potent than that of dopamine and octopamine; the threshold concentrations of epinephrine and norepinephrine were approximately 10(-10) M and those of dopamine and octopamine approximately 10(-7) M. Serotonin weakly inhibited the heartbeat, decreasing both the frequency and amplitude of the myocardial action potential in a concentration dependent manner with a threshold concentration of approximately 10(-6) M. Histamine exhibited no effect on the heartbeat. The results provide the first evidence for direct effects of amines on the crustacean myocardium and suggest neurohormonal regulation of the myogenic heart in T. longicaudatus.     

A change of heart: cardiovascular development in the shrimp Metapenaeus ensis

The larval development of penaeid shrimp is among the most complicated in crustaceans. In Metapenaeus ensis, there are six naupliar, three protozoeal and three mysid larval instars, followed by postlarval development. Irregular heartbeat begins late in naupliar instar 6. Co-ordinated beating at 400-600 beats min(-1) commences in the first protozoeal instar and continues throughout larval life. Initially, the contractile region is located more posteriorly in the cephalothorax and has a single pair of ostia, and the arterial distribution is limited to a single anterior vessel. In later mysid instars, a second cardiac pumping site develops posterior to, but connected with, the original site. This extension is more muscular, contains additional ostia and develops additional distribution vessels supplying the cephalothorax and abdominal areas. The original site is gradually merged into the new extension and only small refinements in the circulation occur in postlarval and juvenile life. Changes in physiological responses of the heart also occur throughout development. Responses to intra-pericardial microinjection of 5-hydroxytryptamine change drastically during development, as do cardiac responses to ambient hypoxia. Similarly, heartbeat of later juvenile instars is inhibited by injection of tetrodotoxin, while heartbeat of larval and early juvenile instars is not, suggesting that neurogenic regulation via the cardiac ganglion arises later in development. Our present studies attempt to integrate the anatomical and physiological changes in the development of the crustacean heart.     

Ontogenetic changes in pacemaker activity in chick heart

Pacemaker activity of the isolated chick heart changed considerably during development. The spontaneous impulse frequency increased up to the 11th–15th incubation day. This period of acceleration was followed by a marked deceleration around the time of hatching. Both acceleration and deceleration appeared to be caused by changes intrinsic to the pacemaker. Deceleration was related to a decrease in rate of slow diastolic depolarization. The results were compared with well-known ontogenetic changes in heart rate $\text{in vivo}$. This comparison indicates that the deceleration of pacemaker firing around hatching is counteracted by the development of sympathetic cardiac reflexes $\text{in vivo}$.     

Beyond Bowditch: the convergence of cardiac chronotropy and inotropy

The ability of the heart to acutely beat faster and stronger is central to the vertebrate survival instinct. Released neurotransmitters, norepinephrine and epinephrine, bind to beta-adrenergic receptors (beta-AR) on pacemaker cells comprising the sinoatrial node, and to beta-AR on ventricular myocytes to modulate cellular mechanisms that govern the frequency and amplitude, respectively, of the duty cycles of these cells. While a role for sarcoplasmic reticulum Ca(2+) cycling via SERCA2 and ryanodine receptors (RyR) has long been appreciated with respect to cardiac inotropy, recent evidence also implicates Ca(2+) cycling with respect to chronotropy. In spontaneously beating primary sinoatrial nodal pacemaker cells, RyR Ca(2+) releases occurring during diastolic depolarization activate the Na(+)-Ca(2+) exchanger (NCX) to produce an inward current that enhances their diastolic depolarization rate, and thus increases their beating rate. beta-AR stimulation synchronizes RyR activation and Ca(2+) release to effect an increased beating rate in pacemaker cells and contraction amplitude in myocytes: in pacemaker cells, the beta-AR stimulation synchronization of RyR activation occurs during the diastolic depolarization, and augments the NCX inward current; in ventricular myocytes, beta-AR stimulation synchronizes the openings of unitary L-type Ca(2+) channel activation following the action potential, and also synchronizes RyR Ca(2+) releases following depolarization, and in the absence of depolarization, both leading to the generation of a global cytosolic Ca(i) transient of increased amplitude and accelerated kinetics. Thus, beta-AR stimulation induced synchronization of RyR activation (recruitment of additional RyRs to fire) and of the ensuing Ca(2+) release cause the heart to beat both stronger and faster, and is thus, a common mechanism that links both the maximum achievable cardiac inotropy and chronotropy.     

Some observations on cardiac automatism in certain animals

Certain aspects of the acetylcholine hypothesis of cardiac automaticity have been tested in vitro with spontaneously beating cardiac tissue from rabbits, rats, dams, and hagfish. The beat of atria from rabbits and rats may be depressed or excited by acetylcholine, depending upon the state of the tissue. Proguanil and cocaine inhibition of the beat in the rat may be antagonized by acetylcholine so that reversal of the depression occurs. The action of acetylcholine on the hearts of clams was found to be strictly inhibitory. Proguanil and cocaine, in contrast to their action on mammalian atria, exert a stimulatory effect on the heart of the molluscs studied. In fact, cocaine stimulated these hearts when they were inhibited by acetylcholine. Studies on the non-innervated hagfish heart revealed that this tissue is completely insensitive to the action of acetylcholine. Extracts prepared from beating hearts of this species will accelerate hypodynamic hearts of the hagfish as well as of the mussel. An extract of the neurogenic lobster heart was without effect on the hagfish heart. Proguanil was likewise ineffective in concentrations which produced inhibition and excitation in rat and clam hearts respectively. It was concluded that acetylcholine does not play a role in the myogenic automatism of all species, and that another mechanism is responsible is suggested on the basis of results obtained in the hagfish hearts.     

A view of the cardiac rhythm control: Intrinsic regulation

Regulation of the cardiac rhythm is intricate and occurs at least at two major levels, intrinsic and extrinsic. In turn, each of these levels can be divided into several sublevels. The factors regulating the cardiac activity eventually affect the duration of spontaneous diastolic depolarization of pacemaker myocytes of the sinoatrial node and, to a far lesser extent, the conduction velocity in the conduction system of the heart. Intrinsic regulation of the heart rate (HR) includes the myogenic sublevel and the sublevels of cell-to-cell communication, the cardiac nervous system, and humoral factors produced within the heart. Myogenic regulation is considered to be the first sublevel in control of the cardiac function. The available data suggest myogenic regulation only for the contractility of the myocardium. The cell-to-cell regulation sublevel involves two principal mechanisms. One depends on the heterogeneous structure of the sinoatrial node and within-node shifts of the dominant pacemaker, which is a group of cells that determine the HR and govern all other cells of the sinoatrial node. The other mechanism is based on the effects of peptides produced by cardiomyocytes and endothelial cells on pacemaker cells of the sinoatrial node. Regulatory peptides are also produced by the cardiac nervous system, which includes sensory and effector autonomic fibers, represents the cardiac part of the metasympathetic system, and forms intramural ganglia. In addition to modulating the HR, these peptides affect the contractility, microcirculation, coronary blood flow, preload, and afterload. Currently available data demonstrate that the autonomic nervous system is far more intricate than believed earlier. Using various neuropeptides, this system provides for fine adjustment of the cell functions, subject to its immediate control.Translated from Fiziologiya Cheloveka, Vol. 31, No. 2, 2005, pp. 116–129.Original Russian Text Copyright © 2005 by Nozdrachev, Kotelnikov, Mazhara, Naumov.Deceased.     

Development of Cardiac Function in Crustaceans: Patterns and Processes

Patterns and mechanisms involved in the onset and developmentof cardiac function in a number of crustacean groups are criticallyreviewed. Irrespective of phylogeny, heart design and ecology,the onset of heart beat seems inextricably linked to the ontogenyof the thoracic segments where the heart is located. Initiallythe beat is erratic but soon becomes regular and the rate increasesas development proceeds. However, still early in developmentthe relationship between heart rate and body size shifts froma positive to a negative one. Nevertheless cardiac output continuesto increase with increasing development, via increasing strokevolume. Some species in more ‘primitive’ groupsdevelop and retain a myogenic heart beat. Others, with globularand tubular hearts, exhibit a shift from myogenicity to neurogenicityaround the time the body size vs. heart rate relationship becomesnegative. Very early cardiac function seems generally insensitiveto external factors, such as temperature, oxygen and pollutants.Sensitivity to environmental factors increases with development,perhaps over the same timescale as the cardiac regulatory mechanismsappear.