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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.

Adrenergic alpha-Agonists↗

Innervation pattern of suboesophageal ventral unpaired median neurones in the honeybee brain.

In honeybees (Apis mellifera), the biogenic amine octopamine has been shown to play a role in associative and non-associative learning and in the division of labour in the hive. Immunohistochemical studies indicate that the ventral unpaired median (VUM) neurones in the suboesophageal ganglion (SOG) are putatively octopaminergic and therefore might be involved in the octopaminergic modulation of behaviour. In contrast to our knowledge about the behavioural effects of octopamine, only one neurone (VUMmx1) has been related to a behavioural effect (the reward function during olfactory learning). In this study, we have investigated suboesophageal VUM neurones with fluorescent dye-tracing techniques and intracellular recordings combined with intracellular staining. Ten different VUM neurones have been found including six VUM neurones innervating neuropile regions of the brain and the SOG exclusively (central VUM neurones) and four VUM neurones with axons in peripheral nerves (peripheral VUM neurones). The central VUM neurones innervate the antennal lobes, the protocerebral lobes (including the lateral horn) and the mushroom body calyces. Of these, a novel mandibular VUM neurone, VUMmd1, exhibits the same branching pattern in the brain as VUMmx1 and responds to sucrose and odours in a similar way. The peripheral VUM neurones innervate the antennal and the mandibular nerves. In addition, we describe one labial unpaired median neurone with a dorsal cell body, DUMlb1. The possible homology between the honeybee VUM neurones and the unpaired median neurones in other insects is discussed.

Animals↗

Forskolin-insensitive adenylate cyclase in cultured cells of Choristoneura fumiferana (Insecta).

Adenylate cyclase from a spruce budworm cell line, IBRI-Cfl, is activated by octopamine (Ka = 50 microM), guanine nucleotides and sodium fluoride but not by forskolin. In addition, forskolin does not potentiate the octopamine-sensitive response. To our knowledge, this is the first published report of a hormone-sensitive adenylate cyclase, with a functional guanine nucleotide regulatory protein, that is insensitive to forskolin both in intact cells and washed membrane preparations.

1-Methyl-3-isobutylxanthine↗

Leydig cells: octopaminergic neurons in the leech.

Leydig cells are electrically-coupled neurons in the segmental ganglia of the leech. This study reports that they stain specifically with high concentrations (0.05 mg/ml) of the dye neutral red, and that they contain octopamine, as demonstrated by radioenzymatic assay. Individual cell bodies were pooled and found to contain 0.37 pmol octopamine/cell body, giving an approximate intracellular concentration of 7.75 mM. Leydig cell bodies contain approximately 75% of the octopamine content of a segmental ganglia. Intracellular injection of large amounts of Lucifer yellow, coupled with long diffusion times, revealed a previously-undescribed Leydig cell process. The probability that Leydig cells are octopaminergic neurosecretory cells is discussed.

Action Potentials↗

Effects of catecholamines, monophenolamines and phenylamines on identifiable giant neurons of an African giant snail (Achatina fulica Férussac).

The effects of catecholamines, monophenolamines and phenylamines on eight identifiable giant neurons of an African giant snail (Achatina fulica Férussac) were examined to classify these neurons into several categories according to their sensitivities to the various substances. The five neurons, PON (periodically oscillating neuron), TAN (tonically autoactive neuron), 1-VMN (left-visceral multiple spike neuron), d-RPeAN (dorsal-right pedal autoactive neuron) and VIN (visceral intermittent firing neuron), were sensitive to catecholamines. Of these neurons, PON was excited most markedly by dopamine (dopamine-sensitive); TAN, 1-VMN and d-RPeAN were inhibited most markedly by epinine (epinine-sensitive); and VIN was excited equally by the four catecholamines, dopamine, epinine, L-noradrenaline and L-adrenaline (widely sensitive). The three other neurons, FAN (frequently autoactive neuron), d-LPeLN (dorsal-left pedal large neuron) and d-LCDN (dorsal-left cerebral distinct neuron) were sensitive to monophenolamines. DL-Octopamine was the most inhibitory on FAN and d-LCDN, but was the most excitatory on d-LPeLN. DL-Synephrine had the same but somewhat weaker effects on the three neurons as did DL-octopamine. The three phenylamines, L-phenylalanine, beta-phenylethylamine and DL-beta-phenylethanolamine, had no effect on any of the eight neurons examined.

2-Hydroxyphenethylamine↗

Effects of catecholaminergic agonists and antagonists on serum gonadotropin concentrations and ovulation in goldfish: evidence for specificity of dopamine inhibition of gonadotropin secretion.

The elevated serum gonadotropin (GtH) levels in goldfish receiving two injections of des Gly10, [D-Ala6] LH-RH ethylamide (LH-RH-A), given 12-hr apart, were reduced by apomorphine, a dopamine agonist, injected at either the first or the second LH-RH-A injection. Serum GtH concentrations in goldfish given two injections of LH-RH-A at a 3-hr interval were also depressed by bromocriptine, a specific D-2 dopamine receptor agonist, administered simultaneously with both LH-RH-A injections. Injections of dopamine antagonists, pimozide or metoclopramide (a specific D-2 antagonist) caused increased serum GtH concentrations in normal goldfish, but no changes were found following injections of the alpha-adrenergic antagonist phentolamine, the beta-adrenergic antagonist propranolol, or the sympathomimetic agent octopamine. Injection of pimozide or metoclopramide at the time of the second of two LH-RH-A injections given at an interval of 12 hr potentiated the LH-RH-A-induced increase in serum GtH concentrations; injections of phentolamine, propranolol, or octopamine did not alter the response to LH-RH-A. Injections of pimozide or metoclopramide also increased the frequency of ovulation in LH-RH-A-injected gravid female goldfish. These results suggest that stimulation of dopamine receptors can block the potentiating effect of multiple injections of GtH-releasing hormone, as well as ongoing LH-RH-A-stimulated release. The results also indicate that the dopamine inhibition of GtH secretion is specific and may be mediated by receptors resembling the D-2 type receptors in mammals.

Animals↗

Down regulation of striatal dopamine receptors in experimental hepatic encephalopathy.

Dopamine receptors were studied in striatal synaptosomes prepared from rat brain with hepatic encephalopathy induced by galactosamine-HCl and documented by visual evoked potential recordings. In order to further characterize the model, plasma amino acid levels and striatal catecholamines and octopamine levels were assayed. In agreement with previous reports in animal and in man, plasma amino acids were increased both in mild and severe stage of this pathology. Striatal levels of norepinephrine and dopamine fell during the development of coma while octopamine rose. Dopamine binding studies showed a decrease in the affinity during the mild stage and a reduction of receptor numbers in the severe stage of encephalopathy. The overall results, in the light of previous reports on GABA receptor studies, seem to indicate the presence in the development of encephalopathy of an imbalance between the dopaminergic and the GABAergic system leading to a prevalence of GABAergic inhibitory system.

Amino Acids↗

Influences of exogenous intake and nitrogen balance on plasma and brain aromatic amino acid concentrations.

In an effort to determine the sum of the factors influencing the plasma and brain concentrations of aromatic amino acids in rats with liver disease, rats with end-to-side portacaval shunts and their sham-operated controls were placed on various oral and intravenous diets by the techniques of parenteral nutrition. In sham-operated animals with normal hepatic function, the source of amino acids and their composition appeared to play little role in the plasma and brain aromatic amino acids and a derivative of brain tyrosine, octopamine. After end-to-side portacaval shunt, however, plasma and brain concentrations of aromatic amino acids were responsive to exogenous intake in the presence of hepatic impairment. In a group of rats with end-to-side portacaval shunt and presumed hepatic impairment, graded increments of protein equivalent were given with isocaloric hypertonic dextrose by the techniques of parenteral nutrition. When the animals achieved positive nitrogen balance, there were decreased levels in plasma and brain of the aromatic amino acids, phenylalanine and tyrosine, and a derivative of tyrosine, octopamine. A clear statistical relationship between plasma and brain tryptophan and nitrogen balance was not observed. The results suggest that in animals with hepatic impairment secondary to end-to-side portacaval shunt both exogenous intake and the level of catabolism (nitrogen balance) influence plasma and brain aromatic amino acids.

Amino Acids↗

Psychophysiological profiles of the Roman strains of rats.

Male rats of the Roman High and Roman Low Avoidance strains were submitted to four principal behavioral tests: food-motivated acquisition of bar-pressing in Skinner box, delayed reinforced alternation, locomotor activity in a "multibox apparatus," conditioned suppression, and two biological measures: blood pressure and brain octopamine level. Performances of RLA and RHA rats were significantly different in each behavioral test. Blood pressure was higher in RHA, but the difference only approached the 0.05 threshold. As previously reported, brain octopamine levels of RHA rats were significantly higher than those of RLA. A multivariate treatment (analysis of correspondences) was applied to the data of 10 behavioral tests. The main factor extracted by this treatment clearly separated the two strains. Among the variables which best differentiate RHA and RLA rats, several do not involve stress (working memory, acquisition of bar-pressing, and locomotor activity).

Animals↗

Properties of an arylalkylamine N-acetyltransferase from the nematode, Ascaridia galli.

1. An arylalkylamine N-acetyltransferase (NAT) of the parasitic nematode, Ascaridia galli was studied using either [14C]serotonin (5-HT) or [14C]octopamine (OA) as substrates and with acetyl-CoA as the donor of the acetate group. 2. The NAT activity towards 5-HT and OA co-eluted from a size-exclusion column and appeared to have an M(r) of around 30,000. The enzyme had apparent Km values of 540 +/- 100 microM (+/- SEM) and 33 +/- 4 microM (+/- SEM) for 5-HT and octopamine, respectively, when assayed in the presence of 1 mM acetyl-CoA. 3. High levels of NAT were found in the gonads of male and female worms and the muscle/body wall. 4. N-acetylation was strongly inhibited by Cu2+ but not by other divalent metal ions and the effect of a number of compounds including biogenic amines, formamidines, hydrazines, and beta-carbolines on the arylalkylamine N-acetyltransferase activity was studied.

Acetyl Coenzyme A↗

Some pharmacological properties of the oviduct muscularis of the stable fly Stomoxys calcitrans.

1. Spontaneous and rhythmic contractions were measured in 80% of the preparations of the stable fly oviduct which were separated from the central nervous system and other tissues. Measurements of the changes in the amplitude and frequency of contractions and changes in the baseline tonus were taken separately, even though they often occurred together during chemical treatments. 2. L-glutamate, at a concentration of 10(-4) to 10(-3) M, caused an increase in the frequency of contractions and in muscle tonus. Contraction frequency showed the highest percentage of change while amplitude and tonus had lower values. 3. The excitation threshold for proctolin on the oviduct was between 10(-13) and 10(-10) M. The initial responses to the peptide showed increases in the amplitude, frequency, and tonus of muscle contractions. However, tonic changes dominated the response profile when the concentration reached 10(-10) or 10(-9) M. In 12 experiments in which the oviduct was exposed to 10(-7) M leucomyosuppressin, no change in the amplitude, frequency, or tonus of muscle contractions was detected. 4. Octopamine caused an inhibition of spontaneous muscle contractions of the oviduct. Spontaneous contractions showed a graded drop in activity over the concentration range tested (10(-8)-10(-6) M). Also 57% of the preparations tested showed a substantial drop in baseline tonus after the addition of octopamine. 5. Stable fly oviducts exposed to extracts from the male reproductive tract showed a noticeable change in the amplitude frequency and tonus of muscle contraction at levels of 2 to 3 equivalents.

Animals↗

Pharmacological sensitivity of the articular capsule of the primary spines of Eucidaris tribuloides.

1. This paper describes the effects of several cholinergic agonists and antagonists, and of beta-phenylethylamine (PEA) and some of its derivatives, on the articular capsule, or ligament, of the primary spines of Eucidaris tribuloides. 2. Carbamylcholine (CCh), methacholine (MeACh), nicotine, and muscarine exert a stiffening effect similar to that of acetylcholine (ACh), although the time course of their actions varies widely. 3. Atropine induced stiffening and blocked and responses to muscarine and MeACh. The responses to MeACh were blocked also by 4-diphenylacetoxy-N-methylpiperidine, suggesting the presence in the ligament of type M3 muscarinic receptors, in addition to nicotinic ones. d-Tubocurarine induced stiffness of the ligament and failed to block the responses to ACh and nicotine. 4. While ACh induced only a slight desensitization, CCh caused a long-lasting blockade of the stiffening effects of the cholinergic agonists. This shows that the receptors for ACh have a site or sites that recognize the ester moieties of these molecules. 5. Eserine and neostigmine potentiate the responses to acetylcholine, indicating the presence of acetylcholinesterase in the ligament. 6. beta-Phenylethylamine, epinephrine, norepinephrine, and dopamine induce diphasic responses; usually a brief softening followed by a slow and irreversible stiffening of the ligament. 7. In contrast to the above, tyramine and octopamine elicit a simple softening of ligaments which are stiff as a result of handling or by exposure to cholinergic agonists. However, tyramine and octopamine do not soften ligaments which become stiff as a result of exposure to adrenergic agonists.

Acetylcholine↗

Polycyclic neuromodulation of the feeding rhythm of the pond snail Lymnaea stagnalis by the intrinsic octopaminergic interneuron, OC.

We have examined the role of the octopamine-containing buccal OC interneuron in the fictive feeding rhythm generated by depolarizing a modulatory interneuron, SO, in the isolated central nervous system (CNS) of Lymnaea stagnalis. Before stimulating the SO, the initial fictive feeding rate was 2.0+/-0.37 bites/min (mean+/-S.E.). When the SO was stimulated, the fictive feeding rate more than doubled, increasing by 5.4+/-2.6 bites/min. Prestimulation of OC facilitates the ability of the modulatory neuron SO to drive fictive feeding 4 s later. Following OC stimulation, the increase in SO-driven feeding rate was 10.8+/-1.6 bites/min, significantly more than when only the SO was stimulated (P<0.02, paired t-test on five preparations). OC activity is not required during the SO stimulation for this enhancement. The maximum of the SO driven rhythm occurs between 6 and 12 s after the end of the OC stimulation at 20 bites/min. This is the maximum feeding rate of intact Lymnaea in sucrose. Facilitation is mimicked by bath applied octopamine at 5 microM. Facilitation is specific to OC interneurons, as the same prestimulation of the electrically coupled neuron N3P (central pattern generator) interneurons does not affect the feeding rhythm. The OC interneuron acts as a long term, polycyclic modulator, which peaks several feeding cycles after the OC activity.

Adrenergic alpha-Agonists↗

High-performance liquid chromatography of biogenic amines in the corpus cardiacum of the American cockroach, Periplaneta americana.

The simultaneous determination of biogenic amines in the corpus cardiacum of the American cockroach, Periplaneta americana, was carried out using high-performance liquid chromatography with a Neurochem neurochemical analyser. Vanillic acid, dopamine, octopamine and tyramine were detected. Tyrosine and tryptophan were also detected at high levels. Octopamine levels in the corpus cardiacum were increased on injection of an acetone solution. The biological function of the biogenic amines detected is discussed.

Animals↗

Heterosynaptic modulation by the octopaminergic OC interneurons increases the synaptic outputs of protraction phase interneurons (SO, N1L) in the feeding system of Lymnaea stagnalis.

We examined the cholinergic synapses between protraction phase interneurons (SO or N1L) and their targets (N1M interneuron, B1 motoneuron) in the buccal ganglia of the pond snail Lymnaea stagnalis. We have tested the hypothesis that the OC (octopamine-containing) interneuron, an intrinsic modulator of the feeding network, can increase the synaptic efficacy from the SO or N1L to their targets. Prestimulation of the OC interneuron, 4 s before the activation of the SO or N1L increases the strength of their output synapses by 75% (SO)-110% (N1L). The individual excitatory postsynaptic potentials evoked by SO or N1L stimulation increase in size. OC prestimulation also produces an increase in the firing rate of these presynaptic interneurons: SO 40%; N1L 33%. The facilitation lasts up to 6 s after the end of the OC burst. The enhancement of PSPs is seen at all the output synapses (both excitatory and inhibitory) of the SO and N1L interneurons. The output synapses of the non-cholinergic swallowing phase N3p interneuron are not affected, even when the same postsynaptic target is selected. The SO-->N1M, SO-->B1 and N1L-->N1M synapses are also strengthened by bath application of 1-5 microM octopamine (average increase 60%). The major effect is an increased excitability of the SO; the B1 motoneuron response to the main transmitter of the SO, acetylcholine, is unaffected. Increased synaptic outputs of the protraction phase SO and N1L interneurons is functionally significant for generation of feeding pattern in the Lymnaea CNS. Strengthening the connections of SO and N1L to the central pattern generator (N1M) interneurons enhances their ability to drive fictive feeding. Thus heterosynaptic facilitation by the octopaminergic OC interneurons in the central pattern generator network may contribute to the behavioral plasticity of feeding in the intact animal.

Acetylcholine↗

A tyramine receptor gene mutation causes a defective olfactory behavior in Drosophila melanogaster.

We characterized molecular profiles of a new olfactory mutant line, honoka (hono), which was found among 500 viable P-element insertion lines screened first by 5-bromo-4-chloro-3-indrolyl-beta-D-galactopyranoside (X-gal) staining on the third segment of the antenna, and then by behavioral assays to several pure chemicals. The behavioral responses of hono mutants to repellents such as ethyl acetate (EA), benzaldehyde (BZ) and 4-methylcycrohexanol (MCH), were reduced compared with those of a control strain. The location of the P-element insertion was determined to be about 100bp) upstream of the first exon of the tyramine receptor gene. The level of 3.6kb tyramine receptor mRNA expression was reduced in hono compared with that of wild-type flies. The tyramine receptor cDNA hybridized to the chromosomal division 79C-D, the same locus as the P-element insertion point in hono, and not to 99A-B, previously reported by Arakawa et al. (1990. Neuron 2, 343-354). Electrophysiological responses to octopamine and tyramine were examined by measuring the excitatory junctional potential (EJP) amplitude from larval body-wall muscles of the hono mutant. The hono was impaired with responding to tyramine, while displaying normal response to octopamine. These results indicate that tyramine has a functional role in the Drosophila olfactory system as a neurotransmitter or a neuromodulator, and hono is the first tyramine receptor mutant. This study provides the first step toward understanding of the molecular genetics of tyramine-mediated neural functions in Drosophila.

Acetates↗

Microassay for N-acetyltransferase activity using high-performance liquid chromatography with electrochemical detection.

A rapid, sensitive procedure has been developed for determination of N-acetyltransferase activity against octopamine, dopamine and 5-hydroxytryptamine. The assay, which is performed in a volume of 10 microliters, is based upon the separation and detection of monoamine substrates and their N-acetylated derivatives using high-performance liquid chromatography with electrochemical detection. The method has been used to measure N-acetyltransferase activity against octopamine, dopamine and 5-hydroxytryptamine in the cerebral ganglion of the American cockroach, Periplaneta americana and to study bi-substrate kinetics of the enzyme.

5-Hydroxytryptophan↗

Cloning of biogenic amine receptors from moths (Bombyx mori and Heliothis virescens).

Based on the similarity of genes which code for guanine-nucleotide binding protein- (G-protein-) coupled receptors, cDNA clones encoding new members of the receptor family have been isolated from Bombyx mori and Heliothis virescens. The deduced protein structures exhibit highest similarity to tyramine/octopamine and serotonin receptors of Drosophila. One of the receptor clones (K50Hel) was permanently expressed in the mammalian cell line LLC-PK1. In stimulation experiments its responded to octopamine leading to an inhibition of adenylate cyclase activity in a dose-dependent manner. Pharmacological studies revealed a higher affinity for mianserin than for yohimbine suggesting, that the K50Hel clone encoded a neuronal type 3 octopamine receptor. As revealed by in situ hybridization, this receptor type is expressed in the central nervous system and antennae of moth.

Amino Acid Sequence↗