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Autoradiographic localization of newly synthesized octopamine to retinal efferents in the Limulus visual system.

The biogenic amine octopamine is synthesized from both tyrosine and tyramine in the lateral, median, and ventral eyes of Limulus. The autoradiographic studies presented here were designed to locate the sites of octopamine synthesis in the ventral and lateral eyes. We found that efferent fibers, which project to ventral and lateral eyes from the central nervous system, became intensely and selectively labeled during in vitro incubations with 3H-tyramine. In the ventral eye, more than 95% of the efferent fibers were labeled. Results of biochemical analyses suggested that most of the radioactive substance within these efferent fibers was newly synthesized octopamine. The selective labeling of efferent fibers during incubation with 3H-tyramine was used as an anatomical tool to study the number and distribution of efferent fibers within the ventral eye. Light microscopic (LM) reconstructions of the distribution of label in serial longitudinal sections through ventral optic nerves together with electron microscopic (EM) autoradiographic analyses revealed between 70 and 200 efferent axons. The results of these studies and of reconstructions of efferent innervation to photoreceptor somata suggest that each ventral photoreceptor cell or each small cluster of cells is innervated by a separate efferent fiber. Both LM reconstructions and EM analyses showed that efferent fibers ramify extensively and specifically in and near the internal rhabdom of ventral photoreceptor cells. In EM autoradiographs of lateral eyes incubated with 3H- tyramine, the silver grains that were located over ommatidia were concentrated exclusively over efferent fibers. All of these efferent fibers, which lay near rhabdoms and in partitions between retinular cells, were labeled. The results of our present studies support our hypothesis that octopamine is a neurotransmitter in Limulus retinal efferent fibers. This amine may modulate the biochemistry and physiology of ventral photoreceptor cells and may mediate many of the known effects of circadian efferent innervation to the lateral eye.

Animals↗

Central nervous sensitization and dishabituation of reflex action in an insect by the neuromodulator octopamine.

Habituation of excitatory synaptic inputs onto identified motor neurons of the locust metathoracic ganglion, driven electrically and by natural stimuli, was examined using intracellular recording. Rapid progressive reduction in amplitude of EPSPs from a variety of inputs onto fast-type motor neurons occurred. The habituated EPSPs were quickly dishabituated by iontophoretic release of octopamine from a microelectrode into the neuropilar region of presumed synaptic action. The zone within which release was effective for a given neuron was narrowly-defined. With larger amounts of octopamine applied at a sensitive site the EPSP became larger than normal, and in many instances action potentials were initiated by the sensitized response. Very small EPSPs onto a motor neuron, which were associated with proprioceptive feedback, and which were originally too small to be detected above the noise, were potentiated to a level of several mV by the iontophoresed octopamine. A DUM neuron (presumed to be octopaminergic) was found, whose direct stimulation was followed by a strong dishabituating and sensitizing action leading to spikes, of inputs to an identified flexor tibiae motor neuron. The action and its time course were closely similar to those evoked by octopamine iontophoresed into the neuropil in the region of synaptic inputs to the motor neuron. It is concluded that DUM (octopaminergic) neurons exert large potentiating actions on central neuronal excitatory synaptic transmission in locusts.

Animals↗

Dopamine- and octopamine-sensitive adenylate cyclase in the brain of adult Culex pipiens mosquitoes.

The effects of dopamine and octopamine on adenylate cyclase activity were studied on the head homogenate of adult Culex pipiens mosquitoes in vitro. Both dopamine and octopamine were shown to increase the cyclic AMP content in the homogenate. The antagonist haloperidol blocked the production of cyclic AMP induced from dopamine but had no effect on the production of cyclic AMP induced by octopamine at the concentrations tested. The opiate agonist etorphine was ineffective at reducing cyclic AMP levels induced by either dopamine or octopamine at the concentrations tested.

Adenylyl Cyclases↗

Octopamine receptor subtypes and their modes of action.

Octopamine receptor subclasses were first proposed to explain differences in the pharmacological profiles of a range of physiological responses to octopamine obtained in the extensor-tibiae neuromuscular preparation of the locust. Thus, OCTOPAMINE1 receptors which inhibit an endogenous myogenic rhythm, increase intracellular calcium levels. Also OCTOPAMINE2 receptors which modulate neuromuscular transmission in this preparation, increase the level of adenylate cyclase activity. The current status of this classification is reviewed by examining the pharmacology of responses to octopamine in a range of preparations. It is concluded that the distinction between OCTOPAMINE1 and OCTOPAMINE2 receptor types is still valid, but that OCTOPAMINE2 receptors exhibit some tissue specific variations. Studies on a cloned Drosophila octopamine/tyramine (phentolamine) receptor are discussed and illustrate many of the difficulties presently encountered in making a definitive classification of octopamine receptors. These include the possibilities that single receptors may activate multiple second messenger systems and that different agonists may differentially couple the same receptor to different second messenger systems.

Animals↗

Octopamine levels during the moult cycle and adult development in the migratory locust, Locusta migratoria.

Octopamine content of the head of the locust Locusta migratoria has been determined during the last larval stage, moulting and adult life of 3 groups of insects: female and male gregarious, solitary and CO2 solitarized. An important difference was found between these 3 groups. Octopamine contents increased in the middle of the larval life and during the adult life. The moulting time is characterized by a sharp decrease of the octopamine content which becomes identical in the 3 groups of insects. The relation between octopamine content, hormone cycles and motility is discussed.

Animals↗

Octopamine, dopamine and noradrenaline content of the brain of the locust, Schistocerca gregaria.

The octopamine, dopamine and noradrenaline content of the brain of the locust, Schistocerca gregaria has been determined using sensitive radiochemical-enzymatic assays. Octopamine and dopamine are present in high concentration but the noradrenaline content is only 1/25 that is octopamine. Both reserpine and fusaric acid (a dopamine-beta-hydroxylase inhibitor) produced a significant depletion of the octopamine stores.

Animals↗

Behavioural and neurochemical effects of intracerebroventricular administrations of p-octopamine in rats.

Intracerebroventricular administration of p-octopamine (250 micrograms) had opposite effects on locomotor activity of rats depending on whether or not the animals were submitted to electric shocks in the experimental situation. When rats were not shocked, their locomotor activity was significantly decreased by the injection. On the other hand, when rats were trained in a shuttle-box, administration of p-octopamine significantly increased avoidance responding and intertrial crossings. The neurochemical effects of the injections were relatively specific: they significantly increased p-octopamine levels in hypothalamus and brainstem but little effect was observed on noradrenaline and dopamine brain contents. These data suggest that octopamine may play the role of a neuromodulator in the central nervous system of mammals.

Animals↗

Octopamine reduces potassium permeability of the glia that form the insect blood-brain barrier.

Octopamine caused only a slight reduction in the potential across the perineurial glia of the cockroach, had no effect upon sodium-induced changes in potential, but did reduce potassium-induced changes (at 10(-7) M and above). The effect of 10(-7) M octopamine was accompanied by a rise in resistance, was mimicked by 10(-7) M synephrine and blocked by 10(-6) M phentolamine. Transperineurial potassium permeability was reduced by 10(-6) M octopamine. It is concluded that octopamine receptors mediate a reduction in potassium conductance of the basolateral membrane of these glia, and a reduction in the net potassium permeability of the barrier.

Animals↗

Colocalization of octopamine and FMRFamide related peptide in identified heart projecting (DUM) neurones in the locust revealed by immunocytochemistry.

Immunocytochemical techniques are employed to reveal colocalization of octopamine with FMRFamide related peptide in the locust ventral nervous system. In each unfused pregenital abdominal ganglia (A4-A6) there are 3 octopamine-like immunoreactive neurones. By combining intracellular Lucifer yellow staining with subsequent immunocytochemistry these are individually identified as the efferent dorsal unpaired median (DUM) neurones DUM-1 and DUM-2, which innervate abdominal tergal and respectively sternal skeletal muscles, and DUM heart-1, an FMRFamide-like immunoreactive neurone which projects to the heart and associated alary muscles. Colocalization of octopamine- and FMRFamide-like immunoreactivity in DUM heart-1 is verified by alternate staining of consecutive sections. With respect to locust ventral ganglia, this investigation shows that colocalization of octopamine with an FMRFamide related peptide is restricted to a single DUM cell occurring in each abdominal ganglion 2-7, which most likely corresponds to segmental homologues of DUM heart-1.

Abdomen↗

Influence of phenylethanolamine on octopamine plasma determination in hepatic encephalopathy.

Octopamine and phenylethanolamine levels were measured by a radioenzymatic procedure in 30 cirrhotic patients with and without hepatic coma and in 15 normal controls. Octopamine data were obtained either by direct extraction with 40% isoamyl alcohol in toluene according to Molinoff et al. (Molinoff, P.B., Landsberg, L. and Axelrod, J. (1969) J. Pharm. Exp. Ther. 170, 253), or after pre-extraction of phenylethanolamine with 3% isoamyl alcohol in toluene. Phenylethanolamine was statistically correlated with the grade of hepatic encephalopathy. Octopamine levels also appeared to parallel the grade of coma, although the values obtained after pre-extraction were lower and less significant than those obtained with 40% isoamyl alcohol in toluene extraction. The higher values of directly extracted octopamine are due to contamination of other beta-hydroxylated phenylethylamines, among which is phenylethanolamine.

2-Hydroxyphenethylamine↗

Noradrenaline-octopamine interactions on cortical neurones in the rat.

The interactions of noradrenaline and octopamine were tested on spontaneously active neurones in the rat cerebral cortex. When applied with weak iontophoretic currents such that no change in baseline firing rate occurred, octopamine profoundly enhanced both depressant and excitatory responses to noradrenaline. Similar applications of octopamine did not alter depressant responses to dopamine or depressant or excitatory responses to iontophoretically applied 5-hydroxytryptamine. The results could support a role of octopamine as a co-transmitter or modulator of noradrenaline mediated neurotransmission.

Animals↗

Isolation and N-terminal amino acid sequence of an octopamine ligand binding protein.

An octopamine receptor photoaffinity probe was used to label membranes from the light organs of Photinus pyralis, a tissue highly enriched in octopamine receptors. Labeling was concentrated in a glycoprotein of 75 +/- 2 kDa with lesser labeling of a 79 +/- 2 kDa component. Labeling could be displaced by prior incubation with octopamine, mianserin, cyproheptadine, phentolamine or propranolol, with a relative potency that correlated with the ability of these same agents to modulate light organ octopamine-sensitive adenylate cyclase. The 75 kDa binding protein was isolated and its N-terminal amino acid sequence was determined.

Affinity Labels↗

The effects of imipramine and iprindole on the metabolism of octopamine in the rat.

Acute injections of imipramine and iprindole in rats produced significant decreases in the concentration of p-hydroxyphenylglycol (pHPG), a neutral metabolite of octopamine in brain at 6 and 24 hr after the administration of drugs. The 24-hr urinary levels of both free and total pHPG were reduced to 25-29% of control with acute administration of imipramine, while iprindole produced a 30% decrease in free pHPG. With chronic administration of imipramine, concentrations of pHPG in brain returned to normal, while the 24-hr urinary levels were still decreased (to 24%). Octopamine in brain was unaltered after both single and repeated injections of imipramine. Thus, these data suggest that the turnover of octopamine in brain is reduced after acute administration of imipramine and iprindole, while after chronic treatment with imipramine, turnover of octopamine in brain has returned to control levels.

Animals↗

Modulation of octopamine-mediated production of cyclic AMP by phorbol-ester-sensitive protein kinase C in an insect cell line.

The presence of protein kinase C (EC 2.7.1.37) in an insect cell line has been demonstrated. Phorbol 12-myristate 13-acetate (PMA), in micromolar concentrations, activated protein kinase C with a translocation of the enzyme from the cytosol to the particulate fraction. Cyclic AMP production in the presence of PMA, octopamine and a combination of both increased in a dose-dependent and time-dependent fashion. The biologically inactive 4 alpha-phorbol 12,13-didecanoate had no effect on protein kinase C activity or on octopamine-mediated cyclic AMP production. Pretreatment of the cells with pertussis toxin had no effect on the response of cells to octopamine or PMA. However, pretreatment with cholera toxin resulted in increased cyclic AMP production which was further enhanced when both cholera toxin and PMA were used in combination. Our data indicate that the octopamine-mediated cyclic AMP production is modulated by protein kinase C.

Animals↗

Analysis of a long-duration hyperpolarization produced by octopamine in an identified effector neuron of Helisoma.

Recent studies have demonstrated that patterned activity in the buccal ganglion of Helisoma trivolvis can be modulated by a variety of neuroactive substances. This study examines the effect of one of these substances, octopamine, on the identified buccal neuron B5. Perfusion of B5 with octopamine produces a 10-20 mV, long-duration hyperpolarization which is associated with an increase in membrane conductance. The magnitude of the hyperpolarization is dose-dependent with a dissociation constant of approximately 5 microM. The reversal potential for the octopamine-induced hyperpolarization (-84 mV) is nearly identical to the predicted potassium equilibrium potential (-85 mV). This result, together with the results of experiments in which extracellular potassium concentrations were altered, demonstrates that octopamine modulated a potassium current in B5.

Animals↗

Prenatal ontogenesis of p-, m-octopamine and phenylethanolamine in relation to catecholamines and their metabolizing enzymes in the developing rat brain and heart.

Non-catecholamines such as phenylethanolamine and p-octopamine are present in many invertebrate nervous systems, sometimes in large amounts. These amines are normally present in the rat brain at much lower levels, p- and m-octopamine are present at trace levels in the mammalian brain. The prenatal development of these amines was studied in comparison with those of noradrenaline and dopamine. The activities of tyrosine hydroxylase, dopa decarboxylase, dopamine beta-hydroxylase and monoamine oxidase were determined in parallel. Phenylethanolamine and p-octopamine are more abundant in the brain between 13 and 17 fetal days than dopamine and noradrenaline but decrease after 17 days whereas the levels of m-octopamine and the two catecholamines increase afterwards. Dopa decarboxylase, dopamine beta-hydroxylase and tyrosine hydroxylase are detected early in fetal life (13, 15 and 14.5 days respectively) but monoamine oxidase activity was not found before 18 days.

2-Hydroxyphenethylamine↗

Octopamine action on the spontaneous contractions of the isolated nerve cord of Lumbricus terrestris.

Octopamine and synephrine were observed to effect the spontaneous rhythmic contractions displayed by the isolated ventral nerve cord of the earthworm, Lumbricus terrestris. octopamine and synephrine produced dose-dependent significant changes in the frequency, amplitude and basal tonus of the spontaneous contractions. Application of adrenergic receptor antagonists suggested the octopamine receptors to have some similarity to vertebrate alpha 1-adrenergic receptors. The spontaneous contractions were not abolished by tetrodotoxin (TTX) which suggested a myogenic origin for the contraction of the ventral nerve cord sheath muscles. Octopamine, in the presence of TTX, increased the basal tonus and maximum force of the spontaneous contractions.

Adrenergic alpha-Antagonists↗

In vitro occurrence of m-octopamine in the cultured cephalic ganglion of Locusta migratoria L. after L-dopa administration.

Cephalic ganglia of the locust Locusta migratoria L. were cultured in the presence of L-DOPA and inhibitors of dopa decarboxylase or dopamine beta hydroxylase. The addition of L-DOPA to the culture medium resulted in a marked increase of m-octopamine, following higher levels of dopamine. L-DOPA produced an increase of m-tyramine. RO 44602 considerably reduced the levels of p and m-octopamines, m-tyramine, dopamine and noradrenaline while fusaric acid reduced only those of p and m-octopamines, m-tyramine and noradrenaline. Though not normally found in the locust nervous system, m-octopamine appears to be closely related to catecholamines and to be produced via their biosynthesizing enzymes through a hypothetical dehydroxylation step.

Animals↗