The effects of small cardioactive peptide B on the isolated heart and gill of Aplysia californica

Effects of small cardioactive peptide B on the physiology of the isolated heart and gill preparations from the mollusc Aplysia californica were examined. In addition, the effects of small cardioactive peptide B and FMRFamide (Phe-Met-Arg-Phe-NH2) on adenylate cyclase activity were compared in particulate fractions of heart and gill tissues, respectively. Small cardioactive peptide B was found to exert dose-dependent, reversible changes in cardiac activity when perfused through the isolated heart. The EC50 values effecting changes in heart rate and force of contraction were 3 X 10(-11) and 3 X 10(-10) M, respectively; minimum concentrations found to effect changes in heart rate and force of contraction were normally 10(-15) and 10(-12) M, respectively. However, some winter hearts demonstrated threshold sensitivity to small cardioactive peptide B at concentrations as low as 10(-17) M. When perfused through the isolated gill, small cardioactive peptide B was found to suppress the gill withdrawal response amplitude with a threshold concentration of 10(-14) M and an EC50 value of 3 X 10(-11) M. Suppression of the gill withdrawal response amplitude by small cardioactive peptide B was found to be dose dependent and reversible up to a concentration of 10(-9) M. At higher concentrations, the suppression tended to persist irreversibly. Small cardioactive peptide B stimulated adenylate cyclase activity in particulate fractions of both heart and gill tissues with an EC50 of 0.1 and 1.0 microM, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Regional, cellular, subcellular distribution of neurospecific protein-hormonal complexes

A sensitive radioimmunological assay (RIA) has been developed to detect the tissue specificity and subcellular localization of three specific protein-hormonal complexes (PHC) of the hypothalamus which have a regulatory function in the brain and visceral organs. Using the highly specific rabbit antisera to beef PHC an order of increasing immunoreactivity in different areas of CNS is as follows: hypothalamus, cerebellum, occipital cortex. The PHC-like immunoreactivity (IR) is found in the neurosecretory granules of the hypothalamo-neurohypophyseal system (about 30%) and in the synaptosomal fraction (70%). In the myelin, mitochondrial and nuclear fractions the PHC-like IR is not revealed. IR of PHC is 1000-fold higher in the brain than in visceral organs (heart, skeletal muscles, adrenals, pancreas and blood serum). A possible role of the PHC as markers of the neuroendocrine cells is discussed.     

Cardioactive effects of hornet venom, Vespa mandarinia

In the experiment of electrocardiogram, the crude venom of giant hornet (Vespa mandarinia) showed cardioactive effects on rat heart. The heart rate was accelerated within 5 min after injection of the venom intraperitoneally, then the heart beat was blocked, resulting in conduction delay. The cardioactive constituent was separated into two components by gel filtration. One which was high molecular species such as protein showed complete atrioventricular block. Another component, having a low molecular weight, was fractionated in 5 peaks, which accelerated heart rate.     

Regulating the activity of a cardioacceleratory peptide

We measured the effect of crustacean cardioactive peptide on Drosophila heart rate in the animal and in a tissue preparation. Crustacean cardioactive peptide increased in vivo basal heart rate 1%, 6%, and 19% and increased in vitro basal heart rate 52%, 25%, and 35% in larvae, pupae, and adults, respectively. In the tissue preparation, the acceleratory period was followed by decreased in vitro heart rates of 42%, 16%, and 13% in larvae, pupae, and adults, respectively. The effects observed in the animal and tissue and in larvae, pupae, and adults suggest that Drosophila crustacean cardioactive peptide cardiac signaling is modulated and developmentally regulated.     

Mathematical model of cardiac mechanogram rhythmicity (based on mollusc heart)

1. The time course of force development by the heart is modelled by Gompertz kinetics from the product of two terms: a cumulative increase in relative number of activated "contractile units", and an exponential decrease in contractile force. 2. For each beat, an "initial condition" is specified by an "intrinsic tension" parameter, and a specific rate of change of tension; cardioactive agents change these specifications. 3. Depending on parameter values, heartbeats are predicted that are constant, or in which the frequency, amplitude and baseline tension are appropriate to inhibited or augmented cardiac activity.     

Peptidergic modulation of cardiovascular dynamics in the Dungeness crab,Cancer magister

Decapod crustacean pericardial organs contain extensive neurohormonal reserves which can be released directly into the haemolymph to act as physiological modulators. The present paper concerns the in vivo effects of two pericardial peptides, proctolin and crustacean cardioactive peptide, on cardiovascular dynamics in the crab Cancer magister. Infusion of proctolin into the pericardial sinus caused a slight decrease in heart rate concurrent with a large increase in cardiac stroke volume. It decreased haemolymph flow anteriorly through the paired anterolateral arteries and increased flow posteriorly and ventrally through the posterior aorta and sternal artery, respectively. The threshold for responses occurred at circulating concentrations of 10^-9 mol·l^-1, and haemolymph flows remained elevated for up to 30 min after peptide infusion. The effects of crustacean cardioactive peptide were less dramatic. Heart rate was not affected but a significant increase in stroke volume was observed. Crustacean cardioactive peptide increased haemolymph flow through the anterolateral arteries and increased scaphognathite rate. The threshold for crustacean cardioactive peptide activity was higher than for proctolin (10^-7 mol·l^-1 and 10^-6 mol·l^-1) but the responses to crustacean cardioactive peptide were of longer duration. The effects of proctolin on regional haemeolymph distribution in Cancer magister closely resemble the cardiovascular responses of this species when exposed to hypoxic conditions. These peptides may be implicated as cardiovascular regulators during environmental perturbations.     

Coronary-active bovine heart proteins induce relaxation in the rabbit aorta]

Seven cardioactive polypeptides have been identified in the precardiac and auricular regions of the bovine heart. Those polypeptides were studied for their action on the function of isolated preparations of the vessel. It has been found that compounds isolated from the precardiac region induce different (20-55%) relaxation of smooth muscles in a strip of the aorta of rabbit. The possibility of formation of endogenic cardioactive compounds in model experiments imitating processing is studied. The data obtained permit assuming and alternatively explaining existence of multiple forms of the mentioned compounds as a result of partial proteolytic splitting of larger molecules.     

Neuroendocrine heart and hypothalamus

Data on the endocrine heart—neurosecretory cells of heart, producing coronarydilatory, metabolically active glycopeptides with physico-chemical and biological properties similar to those of previously discovered cardioactive hypothalamic neurohormones—are summarized. Heart hormones participate in both local and distant regulation of heart metabolism and function. Formation and action of these heart hormones is closely related to hypothalamic cardioactive neurohormones K, C, and G and their protein precursors. Neurohormones from heart and hypothalamus comprise a system of neurohumoral connections between these two organs. A possible role of APUD cells in the generation of a number of heart peptides and glycopeptides exerting hormonal activity is discussed.Abbreviations Used NC Neurohormone C - NK Neurohormone K - NC-H Hypothalamic NC - NK-H Hypothalamic NK - PC-NC NC cleaved from protein-carrier - PC-NK NK cleaved from protein-carrier - BB-NC Brain blood NC - BB-NK Brain blood NK - H₁ Substrate 1 from heart - H₂ Substrate 2 from heart - H₃ Substrate 3 from heart - HB₁ Substrate 1 from heart blood - HB₂ Substrate 2 from heart blood - PC-H₁ H₁ cleaved from protein-carrier - PC-H₂ H₂ cleaved from protein-carrier - PC-H₃ H₃ cleaved from protein-carrier - PC-CC Coronaroconstrictory substance cleaved from protein-carrier - GPG Gomori-positive granules - MLC Mast-like cells     

Cardioactive neuropeptides in gastropods

At least five neuropeptides that are active on an isolated snail heart can be recovered from extracts of gastropod nervous tissue. These peptides have been divided into three classes. The class of the lowest molecular weight, termed the small cardioactive peptides (SCPs), is made up of two peptides. SCPs have been found in all gastropods studied and appear to be involved in the control of the gut. They have been localized by microdissection and bioassay to several identified central neurons that send their axons out to innervate the gut. These neurons act centrally to enhance the motor output of the ganglia responsible for the control of feeding, and peripherally to modulate gut activity. In one pair of these neurons, the classical transmitter acetylcholine coexists with an SCP. The next larger peptide class (medium cardioactive peptide), found only in Aplysia, shares both its mode of cardiac activity and tissue distribution with the SCPs. As yet, there is no evidence that either of these peptide classes acts as a physiological modulator of cardiac activity. The class of the highest molecular weight (large cardioactive peptide [LCPs]) is made up of two peptides and is found only in Helix. The LCPs are circulating neurohormones involved in the regulation of heart, gut, and neuromuscular activity. Their primary release site is a neurohemal region in the auricle. The significance of these findings is discussed in light of recent advances in the study of mammalian neuropeptides.     

Isolation and structure of corazonin, a cardioactive peptide from the American cockroach

Corazonin, a new cardioaccelerating peptide, has been isolated from the corpora cardiaca of the American cockroach, Periplaneta americana, and its structure determined to be Glp-Thr-Phe-Gln-Tyr-Ser-Arg-Gly-Trp-Thr-Asn-amide. The peptide stimulated heart beat at concentrations as low as 0.2 nM, which makes it the most potent insect cardioactive neuropeptide.     

The Role of Small Cardioactive Peptide, SCPB, in the Regulatory Responses of Terrestrial Slugs

The feeding motor program in Limax maximus is the neural correlateof feeding and consists of a discrete pattern of cyclical efferentactivity generated by the buccal ganglia in response to stimulationof chemosensory pathways. The small cardioactive peptide, SCP_B(10^–6 to 10^–9 M), increases the responsiveness ofthe FMP and the endogenous activity of specific feeding motoneuronssuch as the fast salivary burster. Stimulation of buccal neuron,B1, which contains SCP_B-like immunoreactive substance, similarlyincreases the activity of feeding motoneurons. Furthermore,both exogenous SCP_B and stimulation of Bl increase the contractileforce of the heart. Thus it appears that the peptidergic neuronBl is a multifunctional interneuron that is involved in thecontrol of both peripheral and central targets.     

Spin Trapping Evidence for Radical Generation by Isolated Hearts and Cultured Heart Cells

The spin trap 5,5-dimethyl-l-pyrroline-l-oxide (DMPO) has been applied to monitor the generation of free oxy-radicals in samples derived from isolated hearts and heart cells. · OH was trapped in the effluent of isolated hearts in the early phase of reperfusion following an ischemia time of only 10min. Radical detection was possible even when the cardioactive DMPO was added to the effluent after draining off the heart, demonstrating that the short-lived · OH was generated by components released from the affected heart. These results support the hypothesis that radicals are of relevance for reperfusion injury.

By omitting antioxidants commonly used for incubation media of cultured cells, it was possible for the first time to demonstrate the formation of · OH in the incubation solution of cardiac cells.     

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.     

Spin Trapping Evidence for Radical Generation by Isolated Hearts and Cultured Heart Cells

The spin trap 5,5-dimethyl-l-pyrroline-l-oxide (DMPO) has been applied to monitor the generation of free oxy-radicals in samples derived from isolated hearts and heart cells. · OH was trapped in the effluent of isolated hearts in the early phase of reperfusion following an ischemia time of only 10min. Radical detection was possible even when the cardioactive DMPO was added to the effluent after draining off the heart, demonstrating that the short-lived · OH was generated by components released from the affected heart. These results support the hypothesis that radicals are of relevance for reperfusion injury.

By omitting antioxidants commonly used for incubation media of cultured cells, it was possible for the first time to demonstrate the formation of · OH in the incubation solution of cardiac cells.     

Cardioacceleratory action of tachykinin-related neuropeptides and proctolin in two coleopteran insect species

Several cardioactive peptides have been identified in insects and most of them are likely to act on the heart as neurohormones. Here we have investigated the cardioactive properties of members of a family of insect tachykinin-related peptides (TRPs) in heterologous bioassays with two coleopteran insects, Tenebrio molitor and Zophobas atratus. Their effects were compared with the action of the pentapeptide proctolin. We tested the cardiotropic activity of LemTRP-4 isolated from the midgut of the cockroach Leucophaea maderae, CavTK-I and CavTK-II isolated from the blowfly Calliphora vomitoria. The semi-isolated hearts of the two coleopteran species were strongly stimulated by proctolin. We observed a dose dependent increase in heartbeat frequency (a positive chronotropic effect) and a decrease in amplitude of contractions (a negative inotropic effect). In both beetles the TRPs are less potent cardiostimulators and exert lower maximal frequency responses than proctolin. LemTRP-4 applied at 10(-9)-10(-6) M was cardiostimulatory in both species inducing an increase of heart beat frequency. The amplitude of contractions was stimulated only in Z. atratus. CavTK-I and CavTK-II also exerted cardiostimulatory effects in Z. atratus at 10(-9)-10(-6) M. Both peptides stimulated the frequency, but only CavTK-II increased the amplitude of the heart beat. In T. molitor, however, the CavTKs induced no significant effect on the heart.Immunocytochemistry with antisera to the locust TRPs LomTK-I and LomTK-II was employed to identify the source of TRPs acting on the heart. No innervation of the heart by TRP immunoreactive axons could detected, instead it is possible that TRPs reach the heart by route of the circulation. The likely sources of circulating TRPs in these insects are TRP-immunoreactive neurosecretory cells of the median neurosecretory cell group in the brain with terminations in the corpora cardiaca and endocrine cells in the midgut.In conclusion, LemTRP-4, CavTK-I and CavTK-II are less potent cardiostimulators than proctolin and also exert stimulatory rather than inhibitory action on amplitude of contractions. The differences in the responses to proctolin and TRPs suggest that the peptides regulate heart activity by different mechanisms.     

Cardioactive peptides isolated from the brain of a Japanese octopus, Octopus minor

Octopus cardioactive peptides (Ocp-1: Gly-D-Phe-Gly-Asp, and Ocp-3: Gly-Ser-Trp-Asp) were isolated from brain extracts of the octopus, Octopus minor, using the isolated systemic heart as a bioassay. These peptides showed both positive chronotropic and inotropic effects on the heart. The stereoisomers at position 2 were also isolated, but their activities were only 1/10(3)-1/10(4) those of the corresponding isomers. The presence of the peptides in the systemic heart was confirmed by time-of-flight mass spectrometry (MS) and tandem MS analysis. The results suggested that Ocp-1 and Ocp-3 might be involved in excitatory control of the octopus cardiovascular system as neuropeptides and/or neurohormones.     

A cardioactive peptide from the southern armyworm, Spodoptera eridania

A cardioactive peptide was isolated from extracts of whole heads of the southern armyworm, Spodoptera eridania. This peptide has the sequence ENFAVGCTPGYQRTADGRCKPTF (Mr = 2516.8), determined from both Edman sequencing and tandem mass spectrometry in combination with off-line micropreparative capillary liquid chromatography. This peptide, termed Spoer-CAP23, has excitatory effects on a semi-isolated heart from larval Manduca sexta, causing an inotropic effect at low concentrations of peptide and chronotropic and inotropic effects at high doses. The threshold concentration for stimulatory effects of the synthetic peptide on the semi-isolated heart was about 1 nM, suggesting a physiological role as a neuropeptide.     

Multiple polymer architectures of human polyhomeotic homolog 3 sterile alpha motif

The self‐association of sterile alpha motifs (SAMs) into a helical polymer architecture is a critical functional component of many different and diverse array of proteins. For the Drosophila Polycomb group (PcG) protein Polyhomeotic (Ph), its SAM polymerization serves as the structural foundation to cluster multiple PcG complexes, helping to maintain a silenced chromatin state. Ph SAM shares 64% sequence identity with its human ortholog, PHC3 SAM, and both SAMs polymerize. However, in the context of their larger protein regions, PHC3 SAM forms longer polymers compared with Ph SAM. Motivated to establish the precise structural basis for the differences, if any, between Ph and PHC3 SAM, we determined the crystal structure of the PHC3 SAM polymer. PHC3 SAM uses the same SAM–SAM interaction as the Ph SAM sixfold repeat polymer. Yet, PHC3 SAM polymerizes using just five SAMs per turn of the helical polymer rather than the typical six per turn observed for all SAM polymers reported to date. Structural analysis suggested that malleability of the PHC3 SAM would allow formation of not just the fivefold repeat structure but also possibly others. Indeed, a second PHC3 SAM polymer in a different crystal form forms a sixfold repeat polymer. These results suggest that the polymers formed by PHC3 SAM, and likely others, are dynamic. The functional consequence of the variable PHC3 SAM polymers may be to create different chromatin architectures. Proteins 2014; 82:2823–2830. © 2014 Wiley Periodicals, Inc.     

Comparison of the roles of neurohormones in the regulation of blood distribution from the hearts of American and Japanese lobsters

The decapod cardiovascular system consists of a single ventricle that pumps blood into seven arteries; previous work has shown that the outflow distribution patterns of intact animals are variable. In the present study, flow recordings were made from pairs of arteries in semi-isolated hearts whilst different cardioactive hormones were infused into the heart. Each hormone (5-hydroxytryptamine, octopamine, dopamine, proctolin and F1) changed the outflow pattern, heart rate and ventricular pressure in a unique way. The probable sites of hormone action are the cardioarterial valves located at the origin of each artery except one, the dorsal abdominal. Outflow from the dorsal abdominal is controlled downstream by valves located at the origin of the segmental lateral arteries. The responses to a particular hormone were sometimes different between the hearts of American and Japanese lobsters. Accepted: 11 May 1998     

FlyNap (Triethylamine) Increases the Heart Rate of Mosquitoes and Eliminates the Cardioacceleratory Effect of the Neuropeptide CCAP

FlyNap (triethylamine) is commonly used to anesthetize Drosophila melanogaster fruit flies. The purpose of this study was to determine whether triethylamine is a suitable anesthetic agent for research into circulatory physiology and immune competence in the mosquito, Anopheles gambiae (Diptera: Culicidae). Recovery experiments showed that mosquitoes awaken from traditional cold anesthesia in less than 7 minutes, but that recovery from FlyNap anesthesia does not begin for several hours. Relative to cold anesthesia, moderate exposures to FlyNap induce an increase in the heart rate, a decrease in the percentage of the time the heart contracts in the anterograde direction, and a decrease in the frequency of heartbeat directional reversals. Experiments employing various combinations of cold and FlyNap anesthesia then showed that cold exposure does not affect basal heart physiology, and that the differences seen between the cold and the FlyNap groups are due to a FlyNap-induced alteration of heart physiology. Furthermore, exposure to FlyNap eliminated the cardioacceleratory effect of crustacean cardioactive peptide (CCAP), and reduced a mosquito’s ability to survive a bacterial infection. Together, these data show that FlyNap is not a suitable substitute to cold anesthesia in experiments assessing mosquito heart function or immune competence. Moreover, these data also illustrate the intricate biology of the insect heart. Specifically, they confirm that the neurohormone CCAP modulates heart rhythms and that it serves as an anterograde pacemaker.