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PubMed1993

Modulation of identified stomatogastric ganglion neurons in primary cell culture.

1. We studied the properties of identified stomatogastric ganglion (STG) neurons grown in complete isolation in primary cell culture. 2. STG neurons isolated with a short piece of primary neurite adhered to the culture dishes and extended neurites. Outgrowth was apparent within several hours, and continued for < or = 5 days. 3. After 1 day in culture, most STG neurons were not capable of producing action potentials or oscillations. After 3-5 days in culture, most STG neurons regained the ability to fire action potentials, and some became endogenous bursters. Neurons in culture 3-5 days possessed many of the physiological properties of STG neurons in situ, including postinhibitory rebound, a hyperpolarization-activated depolarizing voltage sag, and the ability to burst in the presence of the potassium channel blocker tetraethyl-ammonium. 4. Identified cultured neurons responded appropriately to a variety of neuromodulators, including the monoamines dopamine and octopamine, the muscarinic agonist pilocarpine, and the peptide proctolin. These data suggest that the maintenance of receptor expression in fully differentiated STG neurons is not affected by isolation from all synaptic and modulatory influences. 5. In contrast to the other modulators tested, the effects of serotonin on cultured neurons differed from those reported in situ. Two cell types that are reported to be hyperpolarized by serotonin in situ, the lateral pyloric and pyloric neurons, were depolarized by serotonin in culture.

Action Potentials↗

PubMed1993

Serotonin-containing neurons in lobsters: origins and characterization of inhibitory postsynaptic potentials.

1. The serotonin-containing neurons in the A1 ganglion of the lobster have been shown to act as "gain setters" in neuronal circuits that control the adoption of behaviorally relevant postures. These neurons are subject to tonic inhibition, which has been proposed as an important regulator of their activity. This study explores the pharmacological nature and anatomic location of the neurons responsible for inhibition of these A1 cells; the role played by inhibitory inputs in controlling the firing rates of these neurons is also examined. 2. Three classes of inhibitory postsynaptic potentials (IPSPs) are distinguished in the somata of A1 serotonin-containing neurons. The most common (type I) has amplitudes ranging from 0.4 to 1.5 mV; types II (2-5 mV) and III (< 0.4 mV) are less often seen. 3. Type I IPSPs are reversibly blocked by picrotoxin, a gamma-aminobutyric acid antagonist, but not by serotonin or octopamine antagonists known to act at other lobster synapses. 4. Elimination of type I IPSPs by reversible or irreversible blockage of conduction from the A3 ganglion results in a firing rate increase of approximately 50% in A1 serotonin-containing neurons; IPSP recovery results in a firing rate decrease of corresponding magnitude. Connective transections that do not affect IPSPs do not cause a firing rate increase. 5. Lesions studies suggest that type I IPSPs originate in neurons whose somata are located near the midline of the A3 ganglion; a cell impaled in this region showed action potentials that correlated with IPSPs in an A1 serotonin-containing neuron.

Aggression↗

PubMed1997

Amine modulation of glutamate responses from pyloric motor neurons in lobster stomatogastric ganglion.

The amines dopamine (DA), serotonin (5-HT), and octopamine (Oct) each elicit a distinctive motor pattern from a quiescent pyloric network in the lobster stomatogastric ganglion (STG). We previously have demonstrated that these amines alter the synaptic strength at multiple, distributed sites within the pyloric network that could contribute to the amine-induced motor patterns. Here, we examined the postsynaptic contribution to these changes in synaptic strength by determining how the amines modify responses of pyloric motor neurons to glutamate (Glu), one of the network transmitters, applied iontophoretically into the STG neuropil. Dopamine reduced the Glu responses of the pyloric dilator (PD), ventricular dilator (VD), and inferior cardiac (IC) neurons and enhanced the Glu responses of the lateral pyloric (LP) and pyloric constrictor (PY) neurons. The only effect of 5-HT was to reduce the Glu response of the VD neuron. Oct enhanced the Glu responses of the LP and PY neurons but did not affect the PD, VD, and IC responses. We also examined amine effects on the depolarizing responses to iontophoresed acetylcholine (ACh) in the PD and VD and found that they paralleled the amine effects on Glu responses in these neurons. This suggests that amine modulation of PD and VD responses to Glu and ACh may be explained by general changes in the ionic conductance of these neurons. We compare our results with our earlier work describing amine effects on synaptic strength and input resistance to show that amines act at both pre- and postsynaptic sites to modify graded synaptic transmission in the pyloric network.

Acetylcholine↗

PubMed1999

Long-term effects of prior heat shock on neuronal potassium currents recorded in a novel insect ganglion slice preparation.

Brief exposure to high temperatures (heat shock) induces long-lasting adaptive changes in the molecular biology of protein interactions and behavior of poikilotherms. However, little is known about heat shock effects on neuronal properties. To investigate how heat shock affects neuronal properties we developed an insect ganglion slice from locusts. The functional integrity of neuronal circuits in slices was demonstrated by recordings from rhythmically active respiratory neurons and by the ability to induce rhythmic population activity with octopamine. Under these "functional" in vitro conditions we recorded outward potassium currents from neurons of the ventral midline of the A1 metathoracic neuromere. In control neurons, voltage steps to 40 mV from a holding potential of -60 mV evoked in control neurons potassium currents with a peak current of 10.0 +/- 2.5 nA and a large steady state current of 8.5 +/- 2.6 nA, which was still activated from a holding potential of -40 mV. After heat shock most of the outward current inactivated rapidly (peak amplitude: 8.4 +/- 2.4 nA; steady state: 3.6 +/- 2.0 nA). This current was inactivated at a holding potential of -40 mV. The response to temperature changes was also significantly different. After changing the temperature from 38 to 42 degrees C the amplitude of the peak and steady-state current was significantly lower in neurons obtained from heat-shocked animals than those obtained from controls. Our study indicates that not only heat shock can alter neuronal properties, but also that it is possible to investigate ion currents in insect ganglion slices.

Animals↗

PubMed2001

Amine modulation of the transient potassium current in identified cells of the lobster stomatogastric ganglion.

The pyloric network of the stomatogastric ganglion of the lobster Panulirus interruptus is a model system used to understand how motor networks change their output to produce a variety of behaviors. The transient potassium current (I(A)) shapes the activity of individual pyloric neurons by affecting their rate of postinhibitory rebound and spike frequency. We used two electrode voltage clamp to study the modulatory effects of dopamine (DA), octopamine (OCT), and serotonin (5-HT) on I(A) in the anterior burster (AB), inferior cardiac (IC), and ventricular dilator (VD) neurons of the pyloric circuit. DA significantly reduced I(A) in the AB and IC neurons and shifted their voltages of activation (V(act)) and inactivation (V(inact)) in a depolarized direction. These ionic changes contribute to the depolarization and increased firing rate of the AB and IC neurons produced by DA. Likewise, 5-HT significantly reduced I(A) and shifted V(inact) in the depolarized direction in the IC neuron, consistent with 5-HT's enhancement of IC firing. None of the amines evoked significant changes in I(A) in the VD neuron, suggesting that other currents mediate the amine effects on this neuron.

Algorithms↗

PubMed1978

Presence of hormonally-sensitive adenylate cyclase receptors in capillary-enriched fractions from rat cerebral cortex.

The 10 000 g particulate fraction from capillary-enriched fractions isolated from rat cerebral cortex was shown to possess an adenylate cyclase highly sensitive to activation by sodium fluoride, norepinephrine, epinephrine, isoproterenol and dopamine. To a lesser extent histamine and three dopamine agonists, namely M-7 (5,6-dihydroxy-2-dimethylamino tetralin), ET-495 (methane sulfonate of pyribedil), and S-584 (metabolite of pyribedil) stimulated the enzyme preparation. The action of norepinephrine was blocked by propanolol while phenotolamine and haloperidol were relatively ineffective except at highest concentrations. Phentolamine and propanolol at only highest concentrations (10(-4) M) antagonized the action of dopamine. Haloperidol was seen to be a potent inhibitor of either dopamine- or dopamine agonist-sensitive adenylate cyclase. No effects on the enzyme were observed with methoxamine, octopamine or serotonin. These preliminary data suggest the presence of a mixed population of receptors for adenylate cyclase in rat brain capillaries.

Adenylyl Cyclases↗

PubMed2004

Norepinephrine increases alveolar fluid reabsorption and Na,K-ATPase activity.

The purpose of this study was to determine whether alpha-adrenergic receptor agonists have a role in alveolar fluid reabsorption, via Na,K-ATPase, in the alveolar epithelium. Alveolar fluid reabsorption increased approximately twofold with increasing concentrations of norepinephrine (NE) as compared with control rats. Treatment with the nonselective alpha-adrenergic receptor agonist, octopamine, and the specific alpha(1) agonist, phenylephrine, increased alveolar fluid reabsorption by 54 and 40%, respectively, as compared with control. The specific alpha(1)-adrenergic receptor antagonist, prazosin, inhibited the stimulatory effects of NE by approximately 30%, whereas alpha(2)-adrenergic antagonist, yohimbine, did not prevent the stimulatory effects of NE. The administration of ouabain, Na,K-ATPase inhibitor, prevented the NE-mediated increase in alveolar fluid reabsorption. In parallel with these changes, NE increased Na,K-ATPase activity and protein abundance in alveolar epithelial type II cells via the alpha(1)- and beta-adrenergic receptor. We report here that NE increased alveolar fluid reabsorption via the activation of both alpha(1)- and beta-adrenergic receptors, but not alpha(2)-adrenergic receptors. These effects are due to increased activity and abundance of the Na,K-ATPase in the basolateral membrane of ATII cells.

Adrenergic alpha-Agonists↗

PubMed2003

Neurohormonal and glutamatergic neuronal control of the cardioarterial valves in the isopod crustacean Bathynomus doederleini.

The heart of Bathynomus doederleini gives rise to an anterior median artery (AMA), one pair of anterior lateral arteries (ALAs) and five pairs of lateral arteries (LAs). Cardioarterial valves are located at the junctions between the heart and arteries, each composed of a pair of muscular flaps. All valves of the AMA and the ALAs receive valve excitatory (constrictor) nerves (VEs). The valves of the ALAs receive dual innervation from both constrictor and inhibitor (dilator) nerves, while the valves of the AMA receive innervation from a constrictor nerve alone. The effects of candidate neurohormones on cardioarterial valves were examined by measuring the pressure in each artery at which haemolymph flows out of the heart through the valve. Serotonin, octopamine, norepinephrine, glutamate (Glu) and proctolin constricted the cardioarterial valves and thus decreased the arterial pressure in all the arteries. Dopamine also decreased the arterial pressure of arteries except for the ALAs, in which pressure was increased. Among the neurohormones exerting excitatory effects on the valves, only Glu depolarized the membrane potential of valve muscle cells. The glutamatergic agonists kainate and quisqualate also depolarized the valve muscle cells of the AMA. Excitatory junctional potentials produced in the valves of the AMA in response to the stimulation of a VE were blocked by the glutamatergic antagonists Joro spider toxin and MK-801. Glu is the likeliest candidate for a neurotransmitter for the VEs.

Acetylcholine↗

PubMed2004

Neuronal and neurohormonal control of the heart in the stomatopod crustacean, Squilla oratoria.

The heart of Squilla oratoria contains a cardiac ganglion that consists of 15 intrinsic neurons, supplied by a pair of inhibitory nerves and two pairs of excitatory nerves, arising from the central nervous system. These comprise the extrinsic cardiac innervation. The paired cardio-inhibitor (CI) nerves run out in the 10th pair of nerve roots emerging from the subesophageal ganglion (SEG). The cell bodies of the CI neurons are found in the hemisphere of the 1st segment of the SEG contralateral to the nerve roots in which the CI axons emerge. The two pairs of 1st and 2nd cardio-accelerator (CA1 and CA2) nerves run out in the 16th and 19th pairs of nerve roots of the SEG. The cell bodies of the CA1 and CA2 neurons are found in the hemispheres of the 3rd and 4th segments of the SEG ipsilateral to the nerve roots in which the CA1 and CA2 axons are found. The heartbeat was activated by application of glutamate, serotonin, dopamine, octopamine or acetylcholine, which were applied to the heart by perfusion into an organ bath. Joro-spider toxin (JSTX) blocked myocardial excitatory junctional potentials evoked by the cardiac ganglion. Neuronal cell bodies and processes in the heart were examined using immunocytochemical techniques. All 15 neurons of the cardiac ganglion showed glutamate-like immunoreactivity. Glutamate may be a neurotransmitter of the cardiac ganglion neurons. JSTX also blocked cardiac acceleration by activation of CA1 and CA2 axons. CA1 and CA2 axons showed glutamate-like immunoreactivity. It is likely that glutamate is a neurotransmitter for the cardio-acceleratory neurons. The heartbeat was inhibited by application of gamma-amino-butyric acid (GABA). Cardiac inhibition induced by activation of CI axons was blocked by picrotoxin. CI axons showed GABA-like immunoreactivity. These results may support the identification of GABA as an extrinsic inhibitory neurotransmitter.

Acetylcholine↗

PubMed2006

Beyond the central pattern generator: amine modulation of decision-making neural pathways descending from the brain of the medicinal leech.

The biological mechanisms of behavioral selection, as it relates to locomotion, are far from understood, even in relatively simple invertebrate animals. In the medicinal leech, Hirudo medicinalis, the decision to swim is distributed across populations of swim-activating and swim-inactivating neurons descending from the subesophageal ganglion of the compound cephalic ganglion, i.e. the brain. In the present study, we demonstrate that the serotonergic LL and Retzius cells in the brain are excited by swim-initiating stimuli and during spontaneous swim episodes. This activity likely influences or resets the neuromodulatory state of neural circuits involved in the activation or subsequent termination of locomotion. When serotonin (5-HT) was perfused over the brain, multi-unit recordings from descending brain neurons revealed rapid and substantial alterations. Subsequent intracellular recordings from identified command-like brain interneurons demonstrated that 5-HT, especially in combination with octopamine, inhibited swim-triggering neuron Tr1, as well as swim-inactivating neurons Tr2 and SIN1. Although 5-HT inhibited elements of the swim-inactivation pathway, rather than promoting them, the indirect and net effect of the amine was a reliable and sustained reduction in the firing of the segmental swim-gating neuron 204. This modulation caused cell 204 to relinquish its excitatory drive to the swim central pattern generator. The activation pattern of serotonergic brain neurons that we observed during swimming and the 5-HT-immunoreactive staining pattern obtained, suggest that within the head brain 5-HT secretion is massive. Over time, 5-HT secretion may provide a homeostatic feedback mechanism to limit swimming activity at the level of the head brain.

Adrenergic alpha-Agonists↗

PubMed2006

Division of labor in the honey bee (Apis mellifera): the role of tyramine beta-hydroxylase.

The biogenic amine octopamine (OA) is involved in the regulation of honey bee behavioral development; brain levels are higher in foragers than bees working in the hive, especially in the antennal lobes, and treatment causes precocious foraging. We measured brain mRNA and protein activity of tyramine beta-hydroxylase (T betah), an enzyme vital for OA synthesis, in order to begin testing the hypothesis that this enzyme is responsible for the rising levels of OA during honey bee behavioral development. Brain OA levels were greater in forager bees than in bees engaged in brood care, as in previous studies, but T betah activity was not correlated with bee behavior. T betah mRNA levels, however, did closely track OA levels during behavioral development, and T betah mRNA was localized to previously identified octopaminergic neurons in the bee brain. Our results show that the transcription of this neurotransmitter synthetic enzyme is associated with regulation of social behavior in honey bees, but other factors may be involved.

Amino Acid Sequence↗

PubMed2006

Ancestry of neuronal monoamine transporters in the Metazoa.

Selective Na(+)-dependent re-uptake of biogenic monoamines at mammalian nerve synapses is accomplished by three types of solute-linked carrier family 6 (SLC6) membrane transporter with high affinity for serotonin (SERTs), dopamine (DATs) and norepinephrine (NETs). An additional SLC6 monoamine transporter (OAT), is responsible for the selective uptake of the phenolamines octopamine and tyramine by insect neurons. We have characterized a similar high-affinity phenoloamine transporter expressed in the CNS of the earthworm Lumbricus terrestris. Phylogenetic analysis of its protein sequence clusters it with both arthropod phenolamine and chordate catecholamine transporters. To clarify the relationships among metazoan monoamine transporters we identified representatives in the major branches of metazoan evolution by polymerase chain reaction (PCR)-amplifying conserved cDNA fragments from isolated nervous tissue and by analyzing available genomic data. Analysis of conserved motifs in the sequence data suggest that the presumed common ancestor of modern-day Bilateria expressed at least three functionally distinct monoamine transporters in its nervous system: a SERT currently found throughout bilaterian phyla, a DAT now restricted in distribution to protostome invertebrates and echinoderms and a third monoamine transporter (MAT), widely represented in contemporary Bilateria, that is selective for catecholamines and/or phenolamines. Chordate DATs, NETs, epinephrine transporters (ETs) and arthropod and annelid OATs all belong to the MAT clade. Contemporary invertebrate and chordate DATs belong to different SLC6 clades. Furthermore, the genes for dopamine and norepinephrine transporters of vertebrates are paralogous, apparently having arisen through duplication of an invertebrate MAT gene after the loss of an invertebrate-type DAT gene in a basal protochordate.

Amino Acid Sequence↗

PubMed1985

Neurohormonal modulation of the Limulus heart: amine actions on neuromuscular transmission and cardiac muscle.

The responses of Limulus cardiac neuromuscular junctions and cardiac muscle cells to four endogenous amines were determined in order to identify the cellular targets underlying amine modulation of heartbeat amplitude. The amines increased the amplitude of the Limulus heartbeat, with dopamine (DA) being more potent than octopamine, epinephrine or norepinephrine. The effect of DA on heartbeat amplitude was not blocked by phentolamine. DA enhanced the contractility of deganglionated heart muscle, with time course and dose-dependence similar to its effect on the intact heart. The amines also enhanced neuromuscular transmission, with time course and dose-dependence similar to their effects upon the intact heart. The amplitude of unitary excitatory junction potentials (EJPs) and frequency of miniature excitatory junction potentials (mEJPs) were increased by DA, while mEJP amplitude was unchanged. Thus DA, and probably the other amines, had a presynaptic effect. Combined actions upon cardiac muscle and cardiac neuromuscular transmission account for the ability of these amines to increase the amplitude of the Limulus heartbeat.

Amines↗

PubMed2002

Neurochemical fine tuning of a peripheral tissue: peptidergic and aminergic regulation of fluid secretion by Malpighian tubules in the tobacco hawkmoth M. sexta.

The actions of various peptides and other compounds on fluid secretion by Malpighian tubules in the tobacco hawkmoth Manduca sexta sexta are investigated in this study. Using a newly developed pharate adult Malpighian tubule bioassay, we show that three tachykinin-related peptides (TRPs), leucokinin I, serotonin (5-HT), octopamine, the cardioacceleratory peptides 1a, 1b and 2c, cGMP and cAMP each cause an increase in the rate of fluid secretion in pharate adult tubules. Whereas the possible hormonal sources of biogenic amines and some of the peptides are known, the distribution of TRPs has not been investigated previously in M. sexta. Thus we performed immunocytochemistry using an anti-TRP antiserum. We show the presence of TRP-like material in a small subset of cells in the M. sexta central nervous system (CNS). The larval brain contains approximately 60 TRP-immunopositive cells and there are approximately 100 such cells in the adult brain including the optic lobes. Every ganglion of the ventral nerve cord also contains TRP-like immunoreactive cells. No TRP-containing neurosecretory cells were seen in the CNS, but endocrine cells of the midgut reacted with the antiserum. We propose the hypothesis that the control in insects of physiological systems by hormones may not always involve tissue-specific hormones that force stereotypical responses in their target systems. Instead, there may exist in the extracellular fluid a continuous broadcast of information in the form of a chemical language to which some or all parts of the body continuously respond on a moment-to-moment basis, and which ensures a more effective and efficient coordination of function than could be achieved otherwise.

Adrenergic alpha-Agonists↗

PubMed2002

Drosophila as a new model organism for the neurobiology of aggression?

We report here the effects of several neurobiological determinants on aggressive behaviour in the fruitfly Drosophila melanogaster. This study combines behavioural, transgenic, genetic and pharmacological techniques that are well established in the fruitfly, in the novel context of the neurobiology of aggression. We find that octopamine, dopamine and a region in the Drosophila brain called the mushroom bodies, all profoundly influence the expression of aggressive behaviour. Serotonin had no effect. We conclude that Drosophila, with its advanced set of molecular tools and its behavioural richness, has the potential to develop into a new model organism for the study of the neurobiology of aggression.

Aggression↗

PubMed1976

The actions of some putative neurotransmitters on the cockroach salivary gland.

1. Certain putative transmitters were applied to the innervated cockroach salivary gland and their effects on the resting potential and the neurally evoked secretory potential of the acinar cells were observed. 2. gamma-Aminobutyric acid, glutamate, glycine, aspartate and alanine had no significant effect on the resting potential. However, gamma-aminobutyric acid and glutamate reduced the neurally evoked secretory potential but only at concentrations above 10(-3) M3. Acetylcholine and carbachol appeared to act by modifying transmitter output from the salivary nerves. These substances failed to have any effect on the resting potential. 4. The biogenic amines, adrenaline, dopamine, noradrenaline, 5-hydroxy-tryptamine and octopamine, produced hyperpolarizing responses, graded according to concentration. 5. It is suggested that dopamine, the most potent of the biogenic amines tested, is the transmitter at this junction.

Acetylcholine↗

PubMed1980

Burst reset and frequency control of the neuronal oscillators in the cardiac ganglion of the crab, Portunus sanguinolentus.

1. The five large and four small neurones in the cardiac ganglion of the crab, Portunus, are electrotonically coupled and behave as a single relaxation oscillator, exhibiting periodic bursting activity in vitro. Recorded from the large neurone somata, this activity consists of 200-400 ms slow depolarizations called 'driver potentials' (Tazaki & Cooke, 1979a), accompanied by attenuated action potentials and EPSP's from small neurone input. 2. There is a strong positive correlation between the duration of the driver potential and the duration of the following interburst interval in the spontaneously active ganglion. This correlation is preserved during prolonged depolarization and hyperpolarization. 3. When a driver potential is prematurely terminated by an injected current pulse, the following interburst interval is shortened in direct proportion to the decrease in driver potential duration. 4. When a driver potential or a burst of high-frequency action potential activity is evoked by a depolarizing current pulse, the cardiac oscillator resets to the point of maximum hyperpolarization of the burst cycle, and the following interburst interval is of normal duration. Resetting following an evoked driver potential is complete. Partial resetting occurs only after short, evoked action potential bursts in the absence of a driver potential. 5. Reset of the oscillator causes phase shifts in the subsequent cycles of activity, which vary with the phase of application and duration of the injected current pulse. Response curves have been constructed for a comprehensive range of durations and intensities of hyperpolarizing and depolarizing current pulses applied at all phases of the oscillator cycle. 6. The phase shifts are composed of contributions from the duration of the current pulse, from the premature initiation of the slow depolarizing pacemaker potential due to early termination of the burst, and from the change in interburst interval correlated with truncation of the driver potential. 7. Considering the cardiac ganglion as a relaxation oscillator, frequencey control by entrainment to periodically applied current pulses was quantitatively predicted from the phase-response curves and experimentally confirmed. 8. A high concentration (10(-5) M) of octopamine can inhibit driver potential activity in the large neurones. This was used to examine possible frequency modulating effects of electrotonic feedback from the large neurone driver potentials onto the small neurone pacemaker activity. 9. The observations are discussed in relation to the ionic model for driver potentials and slow pacemaker potential activity in the cardiac ganglion, as proposed by Tazaki & Cooke (1979a, b).

Action Potentials↗

PubMedDate not supplied

An in vitro method for recording the electrical activity of the isolated heart of the adult Drosophila melanogaster.

A recording chamber for monitoring the electrophysiological properties of the isolated heart of adult Drosophila melanogaster has been developed. Spontaneously generated field potentials of constant amplitude can be recorded for 6-8 h (n = 14); in very few cases, records were maintained stable for over 10 h (n = 4), and in some cases below 6 h (n = 5). The chamber consists of the tip of a micropipette, which allows for monitoring the field potential generated by the spontaneously contracting heart. The method can produce accurate information about the heart rate and the amplitude of the cardiac action potential. The preparation can be used for pharmacological studies on the heart of D. melanogaster since it responds, with an increase in the heart rate, to unusually low concentrations of octopamine, 1 nM, a compound with cardioaccelerating properties for insect heart. The recording system can be easily modified for experiments on the heart of other insects. Finally, the isolated heart of D. melanogaster provides a simple method for identifying mutations that affect heart physiology.

Action Potentials↗