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PubMed1987

The metabolism of ingested deuterium-labelled p-tyramine in normal subjects.

Eight normal subjects ingested 125 mg of p-tyramine-beta,beta-2H2 hydrochloride and the 3 h and following 21 h urine collections were analysed by mass spectrometry with selected ion monitoring for the deuterated metabolites: free and conjugated p-tyramine-beta,beta-2H2, free p-octopamine-beta-2H1, free and conjugated p-hydroxyphenylacetic acid-alpha-2H1 and -alpha,alpha-2H2, and free p-hydroxymandelic acid-alpha-2H1. These metabolites accounted for 72% of the ingested label, of which conjugated p-tyramine and free p-hydroxyphenylacetic acid constituted 90%. Approximately 50% of the total deuterated tyramine and 70% of the total deuterated p-hydroxyphenylacetic acid were excreted in the first three hours, although there was considerable variation between individuals. The presence of a small amount of p-hydroxyphenylacetic acid-alpha-2H1 suggests that some exchange of deuterium occurred at the intermediate p-hydroxyphenylacetaldehyde stage. These results, based as they are on metabolites labelled with the stable isotope deuterium, ought to be more reliable than earlier studies in which unlabelled tyramine was ingested with the resultant metabolites, indistinguishable from their endogenous metabolic counterparts, being measured by fluorimetry or gas chromatography.

Administration, Oral↗
PubMed2003

Mesothoracic ventral unpaired median (mesVUM) neurons in the blowfly Calliphora erythrocephala.

The study describes five ventral unpaired median neurons in the mesothoracic neuromere of the fused thoracic ganglion of Calliphora identified by biocytin staining (mesVUM neurons). The group comprises four efferent neurons and one interneuron which are characterized by a common soma cluster in the ventral midline of the neuromere, bifurcating primary neurites and bilaterally symmetrical arborizations. Respective soma clusters of not-yet-identified VUM neurons were also found in the prothoracic, metathoracic, and abdominal neuromeres. The efferent mesVUM neurons are associated with the flight system. Their main arborizations are located in the mesothoracic wing neuropil and their bilateral axons terminate at the flight control muscles, the flight starter muscles, the flight power muscles, or at myocuticular junctions of the latter. In contrast, an association of the interneuron with a particular functional system is not apparent. The arborizations of the neuron are intersegmental and invade all thoracic neuromeres. A further difference between the two types of neurons regards their somatic action potentials, which are overshooting in the efferent neurons and strongly attenuated in the interneuron. Immunocytochemical stainings revealed four clusters of octopamine-immunoreactive (OA-IR) somata in the thoracic ganglion, which reside in the same positions as the VUM somata. We regard this as strong evidence that all groups of VUM neurons contain OA-IR cells and that, in particular, the identified efferent mesVUM neurons are OA-IR. Our results demonstrate that the mesVUM neurons of Calliphora have similar morphological, electrophysiological, and presumably also immunocytochemical characteristics as the unpaired median neurons of other insects.

Animals↗
PubMed2004

Distribution of tyrosine hydroxylase, dopamine, and serotonin in the central nervous system of amphioxus (Branchiostoma lanceolatum): implications for the evolution of catecholamine systems in vertebrates.

To investigate the evolutionary transition that has shaped the catecholaminergic systems of vertebrates, the organization of catecholamine-synthesizing neurons and the nature of the catecholamines were examined in the central nervous system of adult amphioxus (Branchiostoma lanceolatum), a cephalochordate. We isolated a gene transcript encoding tyrosine hydroxylase (TH), the limiting enzyme of catecholamine biosynthesis, and studied its distribution together with that of dopamine and serotonin. Dopamine and TH are found in the same neurons of which they are three separate populations. Two are located in the anterior brain, the first being dorsal and lying in a row and the second being more posterior and lateral. A third population comprising a few dorsal commissural neurons was found in the posterior brain. The anterior dopaminergic cells innervate the ventral commissure of the cephalic vesicle, the hindbrain, and the spinal cord. A serotonin-containing cell group is located in the same plane as the second dopaminergic cell population but is more caudal, marking the probable transition between anterior brain and hindbrain, as deduced from gene expression patterns. The overall distribution of dopaminergic and serotoninergic systems is similar in amphioxus and vertebrate central nervous system and could be an ancestral character of chordates. As assayed by high-performance liquid chromatography and electrochemical detection, significant amounts of dopamine and octopamine, but not of noradrenaline, are present in amphioxus head. This finding is consistent with data obtained from most prostomian species. We conclude that the noradrenergic system is probably an innovation of vertebrates that appeared along with the neural crest and specific hindbrain nuclei.

Amino Acid Sequence↗
PubMed1990

Serotoninergic innervation of the locust mandibular closer muscle modulates contractions through the elevation of cyclic adenosine monophosphate.

The mandibular closer muscles of the locust receive innervation that is immunoreactive for the putative transmitter 5-hydroxytryptamine (5-HT). Cobalt-labelling suggests that the origin of this innervation is a group of cells located anteriorly in the suboesophageal ganglion. Bath application of 5-HT while the muscles are active produces marked changes in the contractions, increasing their amplitude, rate of contraction, and rate of relaxation. Incubation of isolated muscles with 5-HT shows that this amine elevates the levels of the cyclic nucleotide cyclic adenosine monophosphate (cAMP). In addition compounds that artificially elevate the levels of cAMP in the muscle--3-isobutyl-1-methylxanthine (IBMX), forskolin, and the cAMP analogue 8-(4-chlorophenylthio) cAMP--mimic the actions of 5-HT, whereas a potent inhibitor of insect adenylate cyclase, adenosine, considerably delays the onset of the effects produced by 5-HT. The effects observed with 5-HT in the mandibular muscle are similar to those of octopamine in the locust extensor tibiae muscle, and it is possible that this is an analogous modulatory system.

Animals↗
PubMed1993

Fate of abdominal ventral unpaired median cells during metamorphosis of the hawkmoth, Manduca sexta.

Each of the unfused abdominal ganglia in the larval, pupal, and adult stages of the hawkmoth, Manduca sexta, has two large ventral median neurons with axons that bifurcate to innervate targets on both sides of the abdomen. Although the dendritic structures of the two neurons are similar, their axons branch to innervate distinct sets of target muscles. During metamorphosis both neurons undergo dendritic regression, followed by growth of new arborizations during adult development. The neurons must innervate different targets in the larva and adult, since many larval muscles degenerate and are replaced during metamorphosis. Both neurons were reactive with an antibody to the neuromodulatory compound, octopamine, in the larval and adult stages. Pairwise intracellular recordings in isolated nerve cords revealed spontaneous excitatory synaptic potentials that occurred in the ventral median neurons of each ganglion in an anterior-to-posterior sequence. The synaptic potentials were eliminated when the interganglionic connective was interrupted posterior to the subesophageal ganglion. The ventral median neurons were also excited by tactile stimulation of the body surface in larvae, pupae and adults.

Abdomen↗
PubMed2006

Strategies for enantioseparations of catecholamines and structurally related compounds by capillary zone electrophoresis using sulfated beta-cyclodextrins as chiral selectors.

Strategies for simultaneous enantioseparations of three catecholamines (DL-norepinephrine, DL-epinephrine, and DL-isoproterenol) and three structurally related compounds (DL-octopamine, DL-synephrine, and DL-norephedrine) by CZE using sulfated beta-CDs as chiral selectors were investigated. Four different separation modes were attempted: (I) using randomly sulfate-substituted beta-CD (MI-S-beta-CD) at relatively low concentrations in a high-concentration phosphate buffer at low pH in the normal polarity mode, (II) using MI-S-beta-CD at high concentrations at low pH in the reversed polarity mode, (III) using MI-S-beta-CD at moderately high concentrations in a phosphate buffer at neutral pH in the normal polarity mode, and (IV) using the single isomer heptakis(2,3-dihydroxy-6-O-sulfo)-beta-CD (SI-S-beta-CD) at low to moderately high concentrations in a high-concentration BGE at low pH in the normal polarity mode. Among them, enantioseparation of these cationic solutes was best achieved under the conditions of mode (II). In mode (II) and mode (III), temperature is an important factor affecting the enantioresolution of norepinephrine. In mode (I) and mode (IV), the use of a high-concentration BGE (150-200 mM) is crucial for effective enantioseparation of these cationic solutes with sulfated beta-CDs. Comparative studies of enantioseparations of these cationic solutes with MI-S-beta-CD and SI-S-beta-CD reveal that the sulfate substituents of MI-S-beta-CD located at the C(2)- position interact strongly with the diol moiety of catecholamines.

Catecholamines↗
PubMed2002

Neurotransmitters and neuropeptides in the brain of the locust.

As part of continuous research on the neurobiology of the locust, the distribution and functions of neurotransmitter candidates in the nervous system have been analyzed particularly well. In the locust brain, acetylcholine, glutamate, gamma-aminobutyric acid (GABA), and the biogenic amines serotonin, dopamine, octopamine, and histamine most likely serve a transmitter function. Increasing evidence, furthermore, supports a signalling function for the gaseous molecule nitric oxide, but a role for neuroptides is so far suggested only by immunocytochemistry. Acetylcholine, glutamate, and GABA appear to be present in large numbers of interneurons. As in other insects, antennal sensory afferents might be cholinergic, while glutamate is the transmitter candidate of antennal motoneurons. GABA is regarded as the principle inhibitory transmitter of the brain, which is supported by physiological studies in the antennal lobe. The cellular distribution of biogenic amines has been analyzed particularly well, in some cases down to physiologically characterized neurons. Amines are present in small numbers of interneurons, often with large branching patterns, suggesting neuromodulatory roles. Histamine, furthermore, is the transmitter of photoreceptor neurons. In addition to these "classical transmitter substances," more than 60 neuropeptides were identified in the locust. Many antisera against locust neuropeptides label characteristic patterns of neurosecretory neurons and interneurons, suggesting that these peptides have neuroactive functions in addition to hormonal roles. Physiological studies supporting a neuroactive role, however, are still lacking. Nitric oxide, the latest addition to the list of neurotransmitter candidates, appears to be involved in early stages of sensory processing in the visual and olfactory systems.

Animals↗
PubMed1995

Effects of annetocin, an oxytocin-related peptide isolated from the earthworm Eisenia foetida, and some putative neurotransmitters on gut motility of the earthworm.

Annetocin, an oxytocin-related peptide recently isolated from the lumbricid earthworm Eisenia foetida, and putative transmitter substances were examined for their effects on rhythmic, spontaneous contractions of isolated gut preparations of the earthworm. Significant, dose-dependent effects of the following substances were observed: acetylcholine (ACh), gamma-aminobutyric acid (GABA), and dopamine were excitatory, while serotonin (5-HT) and octopamine were inhibitory. Annetocin, oxytocin, and vasotocin stimulated spontaneous contraction of the earthworm gut, annetocin being approximately 10-fold more potent than oxytocin or vasotocin. However, arginine-vasopressin (Arg-vasopressin), lysine-vasopressin (Lys-vasopressin), tocinoic acid (N-terminal hexapeptide fragment of oxytocin), and MSH release-inhibiting factor (MIF; C-terminal tripeptide fragment of oxytocin) did not show any effect on the earthworm gut motility. On the other hand, oxytocin, vasotocin, Arg-vasopressin, Lys-vasopressin, and tocinoic acid caused spontaneous contractions of isolated rat uterine preparations, where the potency was in this order, while annetocin and MIF exerted no oxytocic activity on the uterus. Dose-response relationship of the effects of annetocin and its related peptides on the annelid and mammalian systems shows that amino acid residue at the third position of these peptides is important for exertion of excitatory action on the smooth muscle systems. The results in the present study suggest that receptors for annetocin and for GABA on the earthworm gut, unlike those for ACh, desensitize during continuous exposure to these substances.

Amino Acid Sequence↗
PubMed1989

Sodium-independent transport of noradrenaline in mouse and rat astrocytes in primary culture.

The uptake of noradrenaline by primary cultures of mouse and rat astrocytes was investigated in order to examine whether an inhibition of extraneuronal noradrenaline uptake was the mechanism whereby some trace biogenic amines potentiate neuronal responses to noradrenaline. In the presence of inhibitors of the enzymes monoamine oxidase and catechol-O-methyl transferase, it was found that astrocytes took up noradrenaline by a temperature-dependent, sodium-independent mechanism that was saturable with a Km = 3.4 x 10(-7) M and a Vmax = 1.6 pmole/mg protein/2 min. This uptake mechanism did not concentrate noradrenaline within the cell. The uptake of noradrenaline was inhibited by ascorbic acid (IC50 = 3.4 x 10(-7) M), adrenaline (IC50 = 7.9 x 10(-7) M), and dopamine (IC50 = 1.5 x 1.0(-6) M). It was not inhibited by the tricyclic antidepressants amitriptyline and desmethylimipramine or the trace biogenic amines beta-phenylethylamine, phenylethanolamine, p- and m-tyramine and p- and m-octopamine. Nor was the uptake inhibited by fluoxetine or 5-hydroxytryptamine. It is concluded that astrocytes take up noradrenaline by a facilitated-diffusion mechanism and that this uptake resembles the extraneuronal uptake described in preparations of brain tissue. It is also concluded that the trace biogenic amines do not potentiate neuronal responses to noradrenaline by inhibiting extraneuronal uptake.

Animals↗
PubMed1976

Binding characteristics of drugs to synthetic levodopa melanin.

To define binding characteristics of drugs, levodopa melanin was prepared with the aid of mushroom tyrosinase. The binding of radiolabeled substances was studied with increasing concentrations of melanin in a fixed volume of potassium phosphate buffer (pH 7.4) at 37 degrees. The affinity and capacity of the drug binding were calculated according to Langmuir's adsorption isotherm. The affinity constant of various sympathomimetic amines such as (-)-amphetamine, (+)-amphetamine, (-)-ephedrine, (+/-)-octopamine, and (+)-norepinephrine ranged from 1.1 to 2.8 X 10(5) M-1. The binding capacity for the amines ranged from 1.4 to 3.2 X 10(-9) mole/mg. Although the capacity of (+/-)-cocaine for binding was similar to that of the amines, the affinity was slightly higher, 8.9 X 10(5) M-1. The binding of atropine to the synthetic melanin appeared to be a saturable process with the affinity and capacity values of 0.2 X 10(5) M-1 and 7.6 X 10(-9) mole/mg, respectively. Although the binding lacks stereoselectivity, the drugs vary in their capacity and affinity to bind with melanin. The observed differential pharmacological and toxicologic properties of drugs in the pigmented tissues may in part be related to their differential bi binding characteristics.

Adsorption↗
PubMed1986

Facile synthesis of glycol metabolites of phenethylamine drugs.

High yields of potential glycol metabolites of p-synephrine, epinephrine, octopamine, and normacromerine can be obtained from the readily available monosubstituted and disubstituted acetophenones. The general procedure involves alpha-bromination followed by displacement with acetate ion and reduction with lithium aluminum hydride. Yields ranged from 46 to 91%. Furthermore, the procedure minimizes some problems inherent in aromatic glycol synthesis which include dimerization and pinacol-pinacolone rearrangement.

Chemical Phenomena↗
PubMed1990

Regulation of high affinity choline uptake.

High affinity uptake of choline, the rate-limiting, regulatory step for the synthesis of acetylcholine (ACh), was found to be regulated via presynaptic auto- and heteroreceptors. The transport rate was reduced by a muscarinic agonist and neuropeptides, but was significantly enhanced by octopamine. Intracellular messengers, including cyclic nucleotides, appear to modulate the transport activity, apparently by activating specific protein kinases.

Acetylcholine↗
PubMed1993

Cellular mechanisms governing synaptic development in Drosophila melanogaster.

The neuromuscular connections of Drosophila are ideally suited for studying synaptic function and development. Hypotheses about cell recognition can be tested in a simple array of pre- and postsynaptic elements. Drosophila muscle fibers are multiply innervated by individually identifiable motoneurons. The neurons express several synaptic cotransmitters, including glutamate, proctolin, and octopamine, and are specialized by their synaptic morphology, neurotransmitters, and connectivity. During larval development the initial motoneuron endings grow extensively over the surface of the muscle fibers, and differentiate synaptic boutons of characteristic morphology. While considerable growth occurs postembryonically, the initial wiring of motoneurons to muscle fibers is accomplished during mid-to-late embryogenesis (stages 15-17). Efferent growth cones sample multiple muscle fibers with rapidly moving filopodia. Upon reaching their target muscle fibers, the growth cones rapidly differentiate into synaptic contacts whose morphology prefigures that of the larval junction. Mismatch experiments show that growth cones recognize specific muscle fibers, and can do so when the surrounding musculature is radically altered. However, when denied their normal targets, motoneurons can establish functional synapses on alternate muscle fibers. Blocking synaptic activity with either injected toxins or ion channel mutants does not derange synaptogenesis, but may influence the number of motor ending processes. The molecular mechanisms governing cellular recognition during synaptogenesis remain to be identified. However, several cell surface glycoproteins known to mediate cellular adhesion events in vitro are expressed by the developing synapses. Furthermore, enhancer detector lines have identified genes with expression restricted to small subsets of muscle fibers and/or motoneurons during the period of synaptogenesis. These observations suggest that in Drosophila a mechanism of target chemoaffinity may be involved in the genesis of stereotypic synaptic wiring.

Animals↗
PubMed1995

Diversity and modulation of ionic conductances in leech neurons.

A complete understanding of animal behavior at the cellular level requires detailed information on the intrinsic biophysical properties of neurons, muscles, and the synaptic connections they make. In the past 10 to 15 years, electrophysiological studies of leech neurons have revealed a diverse array of voltage-gated ionic conductances distinguished by their pharmacological sensitivity to classic ion channel blockers. Voltage-clamp studies have provided new information about the kinetics and voltage-dependence of Na+ conductances, several K+ currents, including IA, IK and IK(Ca.), and high- and low-voltage-gated Ca2+ conductances. These studies showed that the action potentials of most leech neurons result from the usual sequence of permeability changes to Na+, K+, and Ca2+ ions. They also added insight as to the role played by particular combinations of conductances in providing individual neurons with electrical properties appropriate for the particular information they encode. Evidence is accumulating on the modulatory actions fo endogenous neurotransmitters such as FMRFamide, serotonin, and octopamine on motor behaviors in the animal. Parallel studies suggest that changes in behavior can be explained, at least in part, by the alteration of firing patterns of selected neurons and muscles resulting from modulation of multiple ion conductances. This makes the leech exceptionally attractive for neuroethological studies because it is one of the simplest organisms in which the methods of psychology and neurobiology can be combined. Information gathered from this animal will therefore increase our understanding regarding general principles underlying the cellular basis of behavior.

Animals↗
PubMed2004

Atmospheric pressure matrix-assisted laser desorption/ionization with analysis by ion mobility time-of-flight mass spectrometry.

The use of an atmospheric pressure matrix-assisted laser desorption/ionization (AP-MALDI) source was employed with an atmospheric pressure ion mobility spectrometer (APIMS) and an orthogonal acceleration reflector time-of-flight mass spectrometer (TOFMS) to analyze dipeptide and biogenic amine mixtures from a liquid glycerol 2,5-dihydroxybenzoic acid (DHB) matrix. Improved sensitivities were obtained by the addition of a localized electrical (corona) discharge in conjunction with the AP-MALDI source. Enhanced sample ionization efficiency created by this combination provided an overall elevation in signal intensity of approximately 1.3 orders in magnitude. Combinations of three dipeptides (Gly-Lys, Ala-Lys, and Val-Lys) and nine biogenic amines (dopamine, serotonin, B-phenylethylamine, tyramine, octopamine, histamine, tryptamine, spermidine, and spermine) were resolved in less than 18 ms. In many cases, reduced mobility constants (K(o)) were determined for these analytes for the first time. Ion mobility drift times, flight times, arbitrary signal intensities, and collision-induced dissociation (CID) fragmentation product signatures are reported for each of the samples.

Atmospheric Pressure↗
PubMed1999

Evidence for adaptive changes in egg laying in crickets exposed to bacteria and parasites.

Animals should increase their present reproductive output if their chances for future reproduction are low. However, an animal's ability to make this adjustment may be constrained by the physiological mechanisms mediating the response. To examine this hypothesis, I infected 2- and 5-week-old female crickets, Acheta domesticus, with either a pathogen (the bacterium Serratia marcescens) that induces antimicrobial immune responses, or a parasite (larvae of the parasitoid fly, Ormia ochracea) that induces an encapsulation immune response. Females of both age groups infected with bacteria laid more eggs the day after injection than did saline-injected crickets. A similar increase was elicited by injecting components of the bacterial cell wall (lipopolysaccharides). The bacteria-induced increase in egg laying (1) was not the result of physical stress, (2) did not appear to be a nonspecific response to the infection, and (3) was probably not mediated by octopamine. Females did not increase egg laying when infested with O. ochracea, even though this parasitoid invariably kills its host. Injections of Sephadex beads, which induced an immune response similar to that created by the parasitoids, also had no effect on egg laying. These results are consistent with the hypotheses that crickets can increase egg laying in response to infection and that increased egg output correlates with the activation of some, but not all, immune responses. Copyright 1999 The Association for the Study of Animal Behaviour.

Journal Article↗
PubMed2000

Cloning of a novel G-protein-coupled receptor from the sea anemone nervous system.

We describe the cloning and analysis of genomic and cDNA copies of a gene from sea anemones that encodes a new member of the G-protein-coupled receptor family. The receptor shows similarity to previously described receptors for biogenic amines such as adrenaline, serotonin, and octopamine, as well as a variety of small molecule agonists and peptides, although we have been unable to determine which ligand is the natural agonist. Antibodies generated against the recombinant receptor protein identify a single protein with a molecular weight of 66 kDa in membrane preparations. Immunofluorescence studies using the same antibody have enabled localization of the receptor in the nervous system. Western blotting and RT-PCR analysis reveal that a homologue of this receptor is expressed in jellyfish and soft coral. We suggest that the receptor plays a role in neurotransmission in the sea anemone and other members of the phylum Cnidaria.

Amino Acid Sequence↗
PubMed1995

Adrenergic and cromolyn sodium modulation of ECL cell histamine secretion.

The histamine secreting enterochromaffin-like (ECL) cell is now recognized as the principal regulator of gastric acid secretion. Histamine is not only a primary modulator of acid secretion, but may be of relevance in gastritis and as a mitogen in gastric neoplasia. Study of the ECL cell has been limited since no pure preparation was available. We therefore developed a pure isolated ECL cell preparation with a purity of 90-95% as determined by total histamine content and chromogranin immunofluorescence. Trypan blue exclusion demonstrated > 95% viability. While gastrin and acetylcholine are known modulators of acid secretion, the role of adrenergic neurotransmitters has not been clearly delineated. The purpose of this study was to examine adrenergic modulation of ECL cell histamine release. To further define the inhibitory mechanisms of histamine secretion, we evaluated the mast cell histamine inhibitor sodium cromoglycate. Histamine secretion was determined by radioimmunoassay. Basal secretion was 0.6 +/- 0.2 nmol/10(3) cells. Gastrin stimulated histamine secretion with an EC50 of 3 x 10(-10) M. Octopamine (alpha-adrenergic agonist) (10(-11)-10(-4) M) failed to stimulate histamine secretion. Isoproterenol (beta-adrenergic agonist) stimulated histamine secretion (EC50, 6 x 10(-8) M) and was inhibited by propranolol (IC50 5 x 10(-10) M).(ABSTRACT TRUNCATED AT 250 WORDS)

Adenylyl Cyclases↗