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Amine and amino acid transmitters in the eye of the mollusc Bulla gouldiana: an immunocytochemical study.

We identified putative transmitters of the photoreceptors and circadian pacemaker neurons and found candidates for efferent control in the eye of the marine mollusc Bulla gouldiana. Established antisera against octopamine, dopamine, serotonin, histamine, glutamate, gamma-aminobutyric acid (GABA), and taurine were used, and central ganglia were processed in parallel to evaluate general staining quality. Photoreceptors and circadian pacemaker cells both expressed immunoreactivity for glutamate and taurine. The eye and its sheath were devoid of GABA-like immunoreactive material, and none of the antisera directed against biogenic amines labelled cells or processes in the nervous tissue of the eye. However, dopamine and octopamine antisera stained large spherical granules (diameter 2-3 microm) contained in granular cells that are located in the connective tissue encapsulating the eye and the optic nerve. The serotonin antiserum revealed a sparse distribution of varicose axon fibers in the optic nerve and eye sheath. No histamine-immunoreactive processes were revealed in the eye. The functional significance of these findings for the molluscan eye and its circadian clock is discussed.

Amines↗

Analysis of the venom of the Sydney funnel-web spider, Atrax robustus using gas chromatography mass spectrometry.

Thirteen compounds have been identified using gas chromatography mass spectrometry in the venom of the Sydney funnel-web spider, Atrax robustus. The compounds were identified as their trimethylsilyl or pentafluoropropionate derivatives and were citric acid, lactic acid, phosphoric acid, glycerol, urea, glucose, gamma-aminobutyric acid, glycine, spermidine, spermine, tyramine and octopamine. Female venom contained trace quantities of 5-methyoxytryptamine which was not detected in male venom. Quantitative determination of tyramine and octopamine was achieved using chemical ionization (CH4) gas chromatography mass spectrometry and deuterated internal standards.

Arthropod Venoms↗

Serotonergic and octopaminergic systems in the squat lobster Munida quadrispina (Anomura, Galatheidae).

Immunocytochemical mapping of serotonergic and octopaminergic neurons in the central nervous system of the squat lobster Munida quadrispina reveal approximately 120 serotonin-immunoreactive cell bodies (distributed throughout the neuromeres except in abdominal ganglion 5) and 48 octopamine-immunoreactive cell bodies (in brain and thoracic neuromeres but none in the circumesophageal or abdominal ganglia). Immunopositive neuropils for both amines are distributed in multiple areas in each neuromere and overlap extensively. Serotonergic and octopaminergic neurons have extensive bilateral projections in abdominal ganglia, whereas the majority of projections in thoracic and subesophageal ganglia are unilateral (contralateral to soma). This difference correlates with typical differences between abdominal and thoracic motor system coordination. Processes of immunoreactive cells for both amines form extensive, peripheral, neurosecretory-like structures. Serotonin seems to be released peripherally in more segments, and from more nerves per segment, than octopamine. M. quadrispina has fewer serotonergic and octopaminergic immunoreactive cells, in particular, fewer segmentally repeated cells, than other species studied to date. Nevertheless, the general organization of the aminergic systems is similar, and several aminergic cells have locations and morphologies that strongly suggest homology with identified aminergic cells in other crustaceans. Among these are segmentally repeated neurons that, in M. quadrispina, form serotonin-immunopositive tubular structures in the thoracic hemiganglia innervating pereiopods 1-3 that are unlike anything reported previously for any species. Comparisons of immunocytochemical maps within one species and between species exhibiting different behaviors provide insights into possible sites of action, functional differences between, and evolution of biogenic aminergic systems.

Animals↗

Enantiomers resolution in capillary zone electrophoresis by using cyclodextrins.

Cyclodextrins added to the background electrolyte are shown to be useful for the resolution of racemic compounds in their enantiomers. Several parameters have to be controlled in order to achieve resolution, e.g., cyclodextrin type, concentration, analyte shape, as well as column temperature. The resolution of nor-epinephrine, epinephrine and isoproterenol in their enantiomers decreased by increasing the column temperature. Octopamine and ketamine have been resolved by supporting the background electrolyte with 2, 6-di-O-methyl-beta-cyclodextrin. In spite of the stronger inclusion-complex of ketamine than octopamine with the modified cyclodextrin its resolution was not satisfactory.

Cyclodextrins↗

Evidence that insect dorsal unpaired medican (DUM) neurons are octopaminergic.

The dorsal unpaired median neuron innervating the extensor tibiae (DUMETi) of locusts and grasshoppers, as well as some other DUM neurons, cause long-term inhibition of the slow intrinsic rhythm of contraction of the muscle by releasing a neurohumoral agent. Large, (600 - 1900 A), dense-core vesicles are present in the nerve terminals. The inhibitory action is mimicked by infusioninto the leg of a minute (0.01 ml.) drop of locust saline containing a very low concentration (10(-6)M) of dopamine or noradrenaline, or an extraordinarily low concentration of octopamine (2.5 x 10(-9) M). The amplitude and frequency of the contractions slowly return to normal. Neither the cell body nor the neuron fluoresces with the Falck-Hillarp treatment, even after a period of ligature has caused an accumulation of vesicles. Octopamine does not fluoresce with this treatment. Accordingly, it is suggested that this neuron, and the other efferent DUM neurons, are octopaminergic.?23Author

Action Potentials↗

Wasp venom injected into the prey's brain modulates thoracic identified monoaminergic neurons.

The wasp Ampulex compressa injects a cocktail of neurotoxins into the brain of its cockroach prey to induce an enduring change in the execution of locomotory behaviors. Our hypothesis is that the venom injected into the brain indirectly alters the activity of monoaminergic neurons, thus changing the levels of monoamines that tune the central synapses of locomotory circuits. The purpose of the present investigation was to establish whether the venom alters the descending control, from the brain, of octopaminergic neurons in the thorax. This question was approached by recording the activity of specific identified octopaminergic neurons after removing the input from the brain or after a wasp sting into the brain. We show that the activity of these neurons is altered in stung and "brainless" animals. The spontaneous firing rate of these neurons in stung and brainless animals is approximately 20% that in control animals. Furthermore, we show that an identified octopamine neuron responds more weakly both to sensory stimuli and to direct injection of current in all treated groups. The alteration in the activity of octopamine neurons is likely to be part of the mechanism by which the wasp induces a change in the behavioral state of its prey and also affects its metabolism by reducing the potent glycolytic activator fructose 2,6-bisphosphate in leg muscle. To our knowledge, this is the first direct evidence of a change in electrical activity of specific monoaminergic neurons that can be so closely associated with a venom-induced change in behavioral state of a prey animal.

Animals↗

Elicitation and abrupt termination of behaviorally significant catchlike tension in a primitive insect.

Sustained steady contractural or catchlike tension (CT) occurs in the metathoracic extensor tibiae muscle of the primitive insect the weta (Orthoptera: Stenopelmatidae) during its characteristic leg-extension defense behavior or following leg-position conditioning. Similar action occurs occasionally in semi-intact preparations and is abruptly turned off by a single peripheral inhibitory impulse. These phenomena were reproduced routinely by first infusing saline containing 10(-8) M (or stronger) octopamine into the muscle for 12 min, and then stimulating the slow excitatory motor neuron SETi with a brief burst. Direct stimulation of the dorsal unpaired median neuron, innervating the extensor tibiae (DUMETi) prior to SETi stimulation, also led to CT. Both octopamine and DUMETi markedly enhanced the tension developed in response to a burst of impulses in SETi.

Animals↗

Identification of novel inhibitors of calling and in vitro [14C]acetate incorporation by pheromone glands of Plodia interpunctella.

Some octopamine agonists were found to suppress in vitro biosynthesis of the calling pheromone of the Indian meal moth, Plodia interpunctella. Isolated pheromone-gland preparations incorporated sodium [14C]acetate at a linear rate for 3 h when incubated with the pheromone biosynthesis activating neuropeptide (PBAN). This incorporation was dependent on the dose of PBAN (up to 0.5 microM). Thin-layer chromatography of a pheromone-gland extract revealed quantitative incorporation of radioactivity into a product exhibiting the same mobility as (Z,E)-9,12-tetradecadienyl acetate, the main component of the calling pheromone of P interpunctella. Twenty-seven octopamine agonists were initially screened using a calling behaviour bioassay of female P interpunctella. Four derivatives with activity in the nanomolar range were identified which were, in order of decreasing pheromonostatic activity: 2-(2,6-diethylphenylimino)thiazolidine > 2-(2,6-diethylphenylimino)oxazolidine > 2-(2,6-dimethylphenylimino)thiazolidine > 2-(2-ethylphenylimino)oxazolidine. These compounds also showed in vitro inhibitory activity in intracellular de novo pheromone biosynthesis. The results of the present study indicate that these derivatives could provide useful information in the characterization and differentiation of octopaminergic receptor types and subtypes.

Adenylyl Cyclases↗

Activation of octopaminergic receptors by essential oil constituents isolated from aromatic plants: possible mode of action against insect pests.

As a result of screening a large number of essential oils from Israeli aromatic plants and their biologically active constituents, we isolated two oils with high activity against several stored-product insects. In this study the effect of these compounds on the acetylcholinesterase and the octopamine systems in insects was studied in order to elucidate their mode of action. Inhibition of acetylcholinesterase activity in vitro was evident only at high concentrations (10(-3) M) and could not account effectively for the low-dose mortality for some stored-product insects observed in vivo. However, the essential oil constituents were found to cause a significant increase in the levels of the intracellular messenger, cyclic AMP of abdominal epidermal tissue in the model insect, Helicoverpa armigera Hübn. The effect was significant even at low, physiological concentrations (10(-8) M) when tested directly on abdominal epidermal tissue preparations in vitro. This intracellular response was found to resemble closely the significant increases in the levels of the cyclic AMP of abdominal epidermal tissue due to treatment with the neurotransmitter/neuromodulator, octopamine. Subsequent treatment with the octopaminergic antagonist, phentolamine, effectively inhibited the cyclic AMP levels induced by essential oil treatment, indicating possible competitive activation of octopaminergic receptors by essential oil constituents.

Acetylcholinesterase↗

Differential modulation of chemical and electrical components of mixed synapses in the lobster stomatogastric ganglion.

1. Two pairs of neurons in the pyloric network of the spiny lobster, Panulirus interruptus, communicate through mixed graded chemical and rectifying electrical synapses. The anterior burster (AB) chemically inhibits and is electrically coupled to the ventricular dilator (VD); the lateral pyloric (LP) and pyloric (PY) neurons show reciprocal chemical inhibition and electrical coupling. We examined the effects of dopamine (DA), serotonin (5HT) and octopamine (Oct) on these mixed synapses to determine the plasticity possible with opposing modes of synaptic interaction. 2. Dopamine increased net inhibition at all three pyloric mixed synapses by both reducing electrical coupling and increasing chemical inhibition. This reversed the sign of the net synaptic interaction when electrotonic coupling dominated some mixed synapses, and activated silent chemical components of other mixed synapses. 3. Serotonin weakly enhanced LP-->PY net inhibition, by reducing electrical coupling without altering chemical inhibition. Serotonin reduced AB-->VD electrical coupling, but variability in its effect on the chemical component made the net effect non-significant. 4. Octopamine enhanced LP-->PY and PY-->LP net inhibition by enhancing the chemical inhibitory component without altering electrical coupling. 5. Differential modulation of chemical and electrical components of mixed synapses markedly changes the net synaptic interactions. This contributes to the flexible outputs that modulators evoke from anatomically defined neural networks.

Animals↗

Dopamine-beta-hydroxylase-like immunoreactive neurons in two insect species, Calliphora erythrocephala and Periplaneta americana.

The localization of dopamine-beta-hydroxylase in the cephalic central nervous system of the blowfly (Calliphora erythrocephala) and the cockroach (Periplaneta americana) was investigated. Immunoreactive neurons were demonstrated in both species. The results were compared with the known distribution of catecholamines in the brain of both species. In certain cell groups and neuropilar regions of both species D beta H-immunoreactivity coincides with the presence of catecholamines. Additionally D beta H immunoreactivity was found in several cell bodies and neuropilar regions in which no catecholamines could be detected. A correlation between the presence of octopamine and anti-D beta H labelling was not found. Thus it seems that the D beta H-immunoreactivity neither indicates the presence of octopamine nor is it limited to noradrenaline-containing neurons. Parallel findings in vertebrates are discussed.

Animals↗

Characterization of the tyraminergic system in the central nervous system of the locust, Locusta migratoria migratoides.

Tyramine occurs in the central nervous system (CNS) of the migratory locust, Locusta migratoria migratoides. The distribution of tyramine within the CNS does not parallel that of octopamine. Tyramine is synthesised from tyrosine in the presence of tyrosine decarboxylase. A second decarboxylase in the CNS is active against 5HTP and DOPA. The locust ganglia incorporate tyramine by high- and low-affinity uptake processes that appear to be independent of dopamine and octopamine. Depolarisation of the locust ganglia by high potassium concentration results in calcium-dependent release of incorporated [3H]tyramine.

Animals↗

Analysis of biogenic amines in plasma of hypertensive patients and a control group.

Procedures were developed for the determination of 17 circulating amines using gas chromatography-negative ion chemical ionisation mass spectrometry. The amines were quantified against their appropriate deuterated isotopomers. The mean concentrations and ranges of catecholamines and trace amines were high compared with previous studies. In comparison with nonhypertensives, plasma from hypertensives had higher concentrations of the following amines: noradrenaline (t = 4.0%); normetanephrine (t = 6.1%) and metanephrine (t = 1.9%). There were no significant differences between 5HT levels in plasma from hypertensives and controls. The following trace amines could be detected in variable amounts in plasma: p-tyramine, m-tyramine, p-octopamine, m-octopamine, p-synephrine, m-synephrine, and salsolinol. The trace amines melatonin, N-acetyl 5HT, tryptamine, 6-hydroxymelatonin and 5-methoxytryptamine could not be detected in plasma with limits of detection lying in the range 20-100 pg ml-1.

Biogenic Amines↗

The inactive mutation leads to abnormal experience-dependent courtship modification in male Drosophila melanogaster.

Flies carrying the inactive mutation of Drosophila melanogaster have only 15% wild-type titers of the putative neurotransmitter octopamine. With a view to discovering whether the inactive mutation impairs learning, I describe the effect of the inactive mutation on experience-dependent courtship modification (EDCM). Wild-type males rapidly condition to immature males and modify their behaviour toward subsequent target flies. The weaker EDCM phenotype of inactive males is similar to that of dunce males. Using time-sampling, habituation is most rapid for wild-type males and slowest for dunce males, with inactive males showing an intermediate phenotype. These results support the notion that octopamine plays a significant role in the manifestation of Drosophila learning behavior.

Animals↗

Effects of biogenic amines and adrenergic drugs on oviposition in the cattle tick Boophilus: evidence for octopaminergic innervation of the oviduct.

Oviposition in the cattle tick Boophilus microplus was arrested by octopamine and the alpha-adrenergic agonists clonidine, tolazoline, and naphazoline, at doses of 25 micrograms per tick. The same effect was obtained with a sublethal dose of Amitraz of 10 micrograms per tick. In contrast, the alpha-adrenergic agonists apomorphine, synephrine, phentolamine, metaclopromide and chlorpromazine were ineffective. No mortality was observed at any of the doses tested. Reserpine, which depletes endogenous adrenergic terminals, mimicked the effects of octopamine in blocking oviposition. Acetylcholine agonists also blocked oviposition but dopaminergic agonists did not. Evidence is presented suggesting that control of the oviduct is mediated by octopaminergic receptors.

Adrenergic Agonists↗

Neuromodulation for behavior in the locust frontal ganglion.

Neuromodulators orchestrate complex behavioral routines by their multiple and combined effects on the nervous system. In the desert locust, Schistocerca gregaria, frontal ganglion neurons innervate foregut dilator muscles and play a key role in the control of foregut motor patterns. To further investigate the role of the frontal ganglion in locust behavior, we currently focus on the frontal ganglion central pattern generator as a target for neuromodulation. Application of octopamine, a well-studied insect neuromodulator, generated reversible disruption of frontal ganglion rhythmic activity. The threshold for the modulatory effects of octopamine was 10(-6) mol l(-1), and 10(-4) mol l(-1) always abolished the ongoing rhythm. In contrast to this straightforward modulation, allatostatin, previously reported to be a myoinhibitor of insect gut muscles, showed complex, tri-modal, dose-dependent effects on frontal ganglion rhythmic pattern. Using a novel cross-correlation analysis technique, we show that different allatostatin concentrations have very different effects not only on cycle period but also on temporal characteristics of the rhythmic bursts of action potentials. Allatostatin also altered the frontal ganglion rhythm in vivo. The analysis technique we introduce may be instrumental in the study of not fully characterized neural circuits and their modulation. The physiological significance of our results and the role of the modulators in locust behavior are discussed.

Action Potentials↗

Effects of neck and circumoesophageal connective lesions on posture and locomotion in the cockroach.

Few studies in arthropods have documented to what extent local control centers in the thorax can support locomotion in absence of inputs from head ganglia. Posture, walking, and leg motor activity was examined in cockroaches with lesions of neck or circumoesophageal connectives. Early in recovery, cockroaches with neck lesions had hyper-extended postures and did not walk. After recovery, posture was less hyper-extended and animals initiated slow leg movements for multiple cycles. Neck lesioned individuals showed an increase in walking after injection of either octopamine or pilocarpine. The phase of leg movement between segments was reduced in neck lesioned cockroaches from that seen in intact animals, while phases in the same segment remained constant. Neither octopamine nor pilocarpine initiated changes in coordination between segments in neck lesioned individuals. Animals with lesions of the circumoesophageal connectives had postures similar to intact individuals but walked in a tripod gait for extended periods of time. Changes in activity of slow tibial extensor and coxal depressor motor neurons and concomitant changes in leg joint angles were present after the lesions. This suggests that thoracic circuits are sufficient to produce leg movements but coordinated walking with normal motor patterns requires descending input from head ganglia.

Action Potentials↗

Modulation of Drosophila heartbeat by neurotransmitters.

The heart of Drosophila melanogaster is a simple muscular tube with a posterior pulsatile portion and a thoracic-cranial vessel. The pacemaker, located caudally, is myogenic. Its rate of firing is modulated by neurotransmitters. Serotonin, octopamine, norepinephrine, dopamine, and acetylcholine accelerate the heart, in that order of potency. Dihydroxyphenylalanine, gamma-aminobutyric acid, glutamate, and glycine have no effect. Generally, the regularity of the heartbeat is not adversely affected by treatment with any of these neurotransmitters. We show here that amnesiac, a neurological mutation, and Dihydroxyphenylalanine decarboxylasetemperature sensitive, a mutation that interferes with synthesis of dopamine, norepinephrine, and serotonin, result in slower heart rate and reduced regularity across a normal range of temperatures for these flies. Dopamine-N-acetyltransferase, which is on the catabolic route to dopamine, serotonin, and octopamine, has no effect. hypoactiveC reduces the rate of the heart, but its mechanism of action is unknown.

Acetylcholine↗