Distribution of [3H]noradrenaline and [3H]serotonin in photophores of Porichthys notatus

Summary Photophores of Porichthys notatus were examined by electron-microscopic radioautography following incubation in tritiated noradrenaline ([³H]NA) or serotonin ([³H]5-HT). Nerve varicosities surrounding the photocytes were found to accumulate [³H]NA but not [³H]5-HT, providing compelling evidence for the catecholaminergic nature of the monoaminergic innervation of photophores. The photocytes themselves appeared selectively labelled with both tracers, but the intensity of labelling after [³H]5-HT incubation was considerably greater than after [³H]NA. Stereological sampling of organelle content in photocytes showed ultrastructural differences between [³H]NA- and [³H]5-HT-labelled cells, probably related to light emission induced by NA. The main changes noted after incubation with [³H]NA were mitochondrial swelling and disorganization, increased coalescence of photocytic vesicles and extrusion of vesicular material into the extracellular matrix. With respect to the subcellular localization of [³H]NA and [³H]5-HT within the photocytes, statistical analysis of the distribution of silver grains disclosed a preferential affinity of both labels for appositional zones between mitochondria and coalescent vesicles. Moreover, in the case of 5-HT, selective affinity was also exhibited by sites comprising vesicular membrane and adjacent cytoplasm, suggesting binding of this biogenic amine to the entire membrane of photocytic vesicles.Supported by grants from the Natural Sciences and Engineering Research Council (M.A.), and Medical Research Council of Canada (L.D.). Dr. Pierre Legendre kindly provided advice on statistical methods     

Co-existence of immunoreactivity to anti-dopamine,anti-serotonin and anti-vasotocin in the cerebral giant neuron of the pond snail Lymnaea stagnalis

Summary Consecutive sections of certain neurons in the central ganglia of the pond snail Lymnaea stagnalis appear to be immunoreactive to anti-dopamine and anti-serotonin. The Cerebral Giant Neurons stain in addition with antivasotocin. The observations indicate the presence of two biogenic amines within the same neuron and in addition their co-existence with a biologically active peptide.     

Interplay of JH, 20E and biogenic amines under normal and stress conditions and its effect on reproduction

Juvenile hormone (JH) and 20-hydroxyecdysone (20E) are well known to play a gonadotropic role in adult insects. In Drosophila the mechanism of reciprocal regulation of JH and 20E is shown to be responsible for their proper balance. Dopamine is a mediator in this JH and 20E interplay. A proper balance between JH and 20E is crucial for the normal progress of oogenesis. An imbalance of gonadotropins leads to reproductive defects: a rise in JH titre leads to oviposition arrest, a rise in 20E level, to the degradation of vitellogenic oocytes. Upon a change in the level of one of the gonadotropins, the balance is restored owing to the relative change in the titre of the other.     

A biochemical characterization of the photophore lenses of the midshipman fish,Porichthys notatus Girard

The present study is a biochemical characterization of the photophore lenses of the midshipman fish, Porichthys notatus, a species that bears 800 photophores distributed over the body surface. The biochemical properties of the photophore lenses were compared with those of the eye lens with which they share a similar developmental origin and analogous function. To achieve a high refractive index, the vertebrate eye lens has a relatively high concentration of structural proteins (20–50%, depending on species) and a simple protein composition, that is, relatively few proteins are synthesized in comparison to other tissues. Similarly, the photophore lenses of P. notatus had a relatively high protein concentration (average = 29%, n = 5) and approximately 60% of the total soluble protein was represented by two subunit species of 33 kD and 35 kD on denaturing polyacrylamide gels. The structural proteins of the eye lens are of two principle types: 1) and polypeptides which belong to vertebrate lens-specific crystallin families, and, 2) enzymes recruited into the lens which take on the function of structural proteins. Here, we report that the two major photophore lens subunits of 33 kD and 35 kD are biochemically similar to each other, but are clearly distinct from any of the previously characterized crystallins. Therefore, we propose that photophore lenses appear to recruit a novel protein.     

Distribution of biogenic amines in the slug,Limax maximus

Summary The distribution of monoamines inLimax maximus was studied by the histochemical fluorescent method of Falck and Hillarp. The number of 5-HT-containing and catecholamine-containing perikarya in the central nervous system is small compared with the non-fluorescent perikarya. However, all the ganglia except the proto-cerebral ganglia have some amine-containing neurons. There are relatively larger numbers of fluorescent cells in the cerebral, visceral, pedal and right parietal ganglia than in the other ganglia. A single, giant 5-HT-containing neuron was observed in each meta-cerebral ganglion.Monoamine neurons are localised in a number of peripheral tissues (heart, integument, tentacles, penis retractor muscle, sole of foot, kidney, alimentary canal, reproductive organs and tentacular, pharyngeal and cephalic retractor muscles). Neurons containing catecholamine are mostly associated with sensory structures such as the statocysts, the retina of the eye and the integument of the tentacles, whereas 5-HT-containing nerve fibres are mainly observed in muscle tissues.We wish to thank the Wellcome Trust for financial support.     

Biogene Amine im Gehirn vom Frosch (Rana esculenta)

Zusammenfassung Mit Hilfe der Methode zur fluoreszenzmikroskopischen Lokalisation von Catechol- und Tryptaminen wurde die Verteilung dieser Stoffe im ZNS von Rana esculenta untersucht. Catecholamin- und serotoninhaltige Neurone liegen im Nucleus reticularis mesencephali. Außerdem finden sich catecholaminhaltige Nervenzellen im Organon vasculosum hypothalami und in der Area praeoptica. Diese aminproduzierenden Zellen entsenden Zellfortsätze durch die Ependymschicht in den Ventrikel. Über diese Ausläufer erfolgt möglicherweise eine Sekretion biogener Amine in den Liquor cerebrospinalis. Catecholamin- und serotoninhaltige Axone erreichen voneinander verschiedene Kerngebiete und Areale. Neben dem periventrikulären Zellager im Tuber cinereum und in der Area praeoptica werden vor allem der ventrolaterale Teil des lateralen Septumkerns, Striatum ventrale und Epistriatum von Endstrecken catecholaminhaltiger Axone durchdrungen. Serotoninhaltige Varicositäten finden sich dagegen vor allem in Kerngebieten, die in sensorische Bahnen eingeschaltet sind (Nucleus isthmi, corpus geniculatum laterale, Area praetectalis, Tectum opticum, Thalamus dorsalis, Neostriatum). Weitere Ausbreitungsgebiete 5-Hydroxytryptamin-haltiger Fasern sind die Habenula und der Nucleus interpeduncularis, Kerngebiete, über die Erregungen aus dem limbischen System auf vegetative Zentren der Medulla oblongata geleitet werden.

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Biogenic amines in the brain of the frog (Rana esculenta)

Summary The distribution of biogenic amines in the central nervous system of Rana esculenta was investigated by means of the fluorescence-microscopical detection of catecholand tryptamines. The nucleus reticularis mesencephali was found to contain numerous neurones rich in catechol- and tryptamines. Apart from this nucleus nerve cells in the organon vasculosum hypothalami and in the area praeoptica were found to contain catecholamines. The clublike processes of these neurones penetrate the ependymal layer and extend into the ventricle. These structures are presumably responsible for a secretion of biogenic amines into the cerebrospinal fluid. Catecholamine- and serotonin-containing axons terminate on different nuclei and areas. Besides the periventricular cellular layer of the tuber cinereum and the area praeoptica, the pars ventrolateralis of the nucleus septalis lateralis, striatum ventrale and epistriatum are pervaded by terminals of catecholamine-containing neurons. Serotonincontaining varicosities are mainly to be found in nuclei, which are intercalated in sensory pathways (nucleus isthmi, corpus geniculatum laterale, area praetectalis, tectum opticum, thalamus dorsalis, neostriatum). Further areas of distribution of 5-hydroxytryptamine-fibers are the habenula and the nucleus interpeduncularis, nuclei which coordinate impulses from the limbic system projecting them on visceral centers of the medulla oblongata.

Mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Factors affecting tyramine production in <Emphasis Type="Italic">Enterococcus durans</Emphasis> IPLA 655

The decarboxylation of tyrosine by certain lactic acid bacteria leads to the undesirable presence of tyramine in fermented foods. Tyramine is the most frequent biogenic amine found in cheese and is also commonly found in other fermented foods and beverages. The tyramine-producing strain Enterococcus durans IPLA 655 was grown in a bioreactor under different conditions to determine the influence of carbon source, tyrosine and tyramine concentrations, and pH on tyramine production. The carbon source appeared to have no significant effect on the production of tyramine. In contrast, tyrosine was necessary for tyramine production, while the presence of tyramine itself in the growth medium inhibited such production. pH showed by far the greatest influence on tyramine synthesis; tyramine was produced in the greatest quantities at pH 5.0, although this was accompanied by a reduced growth rate.     

Adrenergic control of hypothalamic function during osmotic stress in the mallard duck (Anas platyrhynchos)

Summary The role of the paraventricular nucleus (PVN) and biogenic amines (BA) in regulating the level of corticoids in the serum of osmotically stressed mallard ducks (Anas platyrhynchos) was analyzed employing three experimental approaches: 1) pharmacologic alteration of central BA levels, 2) microscopic evaluation of BA distribution, and 3) placement of electrolytic lesions into the PVN. Reserpine and -methyl-p-tyrosine (mpt), agents that decrease the amount of BA's in the central nervous system, produced a fivefold increase in the concentration of serum corticoids. Conversely, pargyline and amphetamine, agents that increase the functional pool of BA's, prevented the rise in serum corticoid concentration normally observed in birds challenged with an intraperitoneal injection of hypertonic saline. When the topographic distribution of BA's was analyzed in the brains of osmotically stressed and nonstressed ducks distinct changes in the intensity of catecholamine (CA) fluorescence were observed in only one location, the PVN of the hypothalamus. Additionally, electrolytic lesions stereotaxically placed in the PVN blocked the osmotic stress-induced rise in serum corticoid concentration. These data therefore indicate that the PVN in the mallard duck plays some role in regulating the observed stress-induced rise in serum corticoid concentration, and that this regulatory function is probably inhibited by catecholamines.This research was supported by research grant No. GB 33321 from the National Science Foundation. We wish to express our sincere thanks to Mr. Howard Funk, research director, Colorado Division of Wildlife, for the use of the State's animal facilitiesThis research was submitted as partial fulfillment for the degree of Doctor of Philosophy, Department of Physiology and Biophysics, Colorado State University, Ft. Collins, CO 80521     

Immunocytochemistry of histamine in the brain of the locust<Emphasis Type="Italic"> Schistocerca gregaria</Emphasis>

Histamine serves a neurotransmitter role in arthropod photoreceptor neurons, but is also present in a small number of interneurons throughout the nervous system. In search of a suitable model system for the analysis of histaminergic neurotransmission in insects, we mapped the distribution of histamine in the brain of the desert locust Schistocerca gregaria by immunocytochemistry. In the optic lobe, apparently all photoreceptor cells of the compound eye with projections to the lamina and medulla showed intense immunostaining. Photoreceptors of the dorsal rim area of the eye had particularly large fiber diameters and gave rise to uniform varicose immunostaining throughout dorsal rim areas of the lamina and medulla. In the locust midbrain 21 bilateral pairs of histamine-immunoreactive interneurons were found, and 13 of these were reconstructed in detail. While most neuropil areas contained a dense meshwork of immunoreactive processes, immunostaining in the antennal lobe and in the calyces of the mushroom body was sparse and no staining occurred in the pedunculus and lobes of the mushroom body, in the protocerebral bridge, and in the lower division of the central body. A prominent group of four immunostained neurons had large cell bodies near the median ocellar nerve root and descending axonal fibers. These neurons are probably identical to previously identified primary commissure pioneer neurons of the locust brain. The apparent lack in the desert locust of certain histamine-immunoreactive neurons which were reported in the migratory locust may be responsible for differences in the physiological role of histamine between both species.The study was supported by the Deutsche Forschungsgemeinschaft, grants Ho 950/13 and 950/14     

Comparing injection, feeding and topical application methods for treatment of honeybees with octopamine

Entomologists have used a range of techniques to treat insects with neuroactive compounds, but it is not always clear whether different treatment methods are equally effective in delivering a compound to a target organ. Here, we used five different techniques to treat honeybees with 3H-octopamine (3H-OA), and analysed the distribution of the 3H radiolabelled compound within different tissues and how it changed over time. All treatment methods, including injection of the median ocellus, resulted in 3H-OA detection in all parts of the honeybee. Injection through the median ocellus was the most effective method for delivering 3H-OA to the brain. Topical application of 3H-OA dissolved in dimethylformamide (dMF) to the thorax was as effective as thoracic injections of 3H-OA in delivering 3H-OA to the brain, but topical applications to the abdomen were less so. Most of the 3H-OA applied topically remained associated with the cuticle and the tissues of the body segment to which it had been applied. For all treatment methods, 3H-OA was rapidly lost from the brain and head capsule, and accumulated in the abdomen. Our findings demonstrate the value of thoracic topical treatment with compounds dissolved in dMF as an effective non-invasive method for short-term, systemic pharmacological treatments.     

The taxonomic position of Cyanidium,Cyanidioschyzon and Galdieria: an update

The ecophysiological, cytomorphological, biochemical and molecular data presently available for the acidophilic red algal species Cyanidium caldarium, Cyanidioschyzon merolae and Galdieria sulphuraria are summarised. The taxonomic position of the three genera is discussed and emendements to the generic diagnosis are presented.     

Regional Variations of 5HT Concentrations in Rora sg (staggerer) Mutants

Ataxic Rora ^sg (staggerer) mouse mutants, containing a deletion of the Rora gene which encodes a retinoid-like nuclear receptor, were compared to non-ataxic controls for concentrations of 5-hydroxytryptamine (HT), its main metabolite (5-hydroxy-indole acetic acid, 5HIAA), and its precursor (tryptophan) in cerebellum, brainstem, and forebrain. In Rora ^sg cerebellum, 5HT concentrations increased relative to controls, while tryptophan concentrations decreased. 5HIAA concentrations increased in mutant cerebellum and brainstem, but the 5HIAA/5HT ratio declined only in cerebellum. These results indicate that 5HT turnover decreased in cerebellum of an ataxic mutant, perhaps indicative of presynaptic accumulation and compromised neurotransmission and susceptible to be modified by 5HT pharmacotherapy.     

Taxonomic and functional implications of stigma morphology in species ofCassia,Chamaecrista, andSenna (Leguminosae:Caesalpinioideae)

Two stigma forms occur inChamaecrista andSenna, but only one inCassia. In the common chambered form, a stigma pore is positioned on the reflexed style tip and is the entrance to a tapering chamber. The pore rim is fringed by hairs which vary in number, size, distribution and shape. In the alternative form the stigma is situated at the apex of the curved style and is crateriform. The crater rim is fringed by hairs of variable number and shape. The stigmatic hairs are predominantly unicellular and cutinized. Stigma and hair differences aid in the taxonomy of the genera. Their functions in pollination biology are discussed.     

Catecholamines in the coelenterate Renilla köllikeri

Summary The characteristics of uptake of ³H-noradrenaline (³H-NA) and ³H-adrenaline (³H-A) in the tissues of the sea pansy, Renilla köllikeri, were studied by in vivo incubations. Lineweaver-Burk plots indicated two components of catecholamine accumulation, one representing a high-affinity uptake with an apparent K _m of 4.91×10^-7 M (³H-NA) or 4.39×10^-7 M (³H-A), and the other a low affinity process with an apparent Km of 5.52×10^-5 M (³H-NA) or 1.49×10^-5 M (³H-A). The high-affinity uptake of both tracers was strongly inhibited at low temperature and in a calcium-free medium, thus suggesting the involvement of a carrier-mediated transport mechanism, but was largely insensitive to sodium omission and ouabain. Accumulations of ³H-NA, but not ³H-A, were highly desipramine-sensitive.Light-microscopic radioautographic studies demonstrated the presence of cells reactive to both ³H-NA and ³H-A in the ectoderm, mesoglea and endoderm. Extraneuronal accumulations of ³H-NA and ³H-A were prominent in some ectodermal cells, in amoebocytes and spicule cells. Reactive neuronal processes were tentatively identified throughout the mesoglea and over all muscle layers on the basis of several morphological criteria. ³H-A, but not ³H-NA label, was more intense over the presumed photocytic zone and circular muscle than elsewhere. These and other observations support a neurotransmitter role for adrenaline (and probably noradrenaline) in control of luminescence and modulation of slow rachidial contractions.Supported by a grant from the Natural Sciences and Engineering Research Council of Canada     

Étude des Amines Biogènes de Glycera convoluta K. (Annélide Polychète)

Résumé L'étude par la méthode histochimique de fluorescence de Falck, du ganglion cérébroïde ét de la chaine nerveuse ventrale de Glycera convoluta K. a permis la mise en évidence de péricaryons et de fibres présentant une fluorescence jaune et verte. La nature et le métabolisme des amines mises en évidence histochimiquement ont été étudiées par des techniques biochimiques. La présence d'adrénaline et de 5-hydroxytryptamine a pu être établie avec certitude. Deux autres amines identifiées chromatographiquement comme étant de la dopamine et de la noradrénaline présentent cependant des caractéristiques spectrofluorimétriques légèrement différentes de ces deux substances.La rétention de la noradrénaline et de la dopamine étudiée par des techniques radiochimiques s'est montrée plus longue que dans le canal déférent du Rat. Les composants lipidiques, protéolipidiques et protéiques étudiés chromatographiquement sont apparus assez différents de ceux rencontrés chez les Mammifères.Des données récentes ont montré l'importance de l'arrangement spatial des groupements fonctionnels dans la molécule du médiateur par rapport à la (ou aux) protéine (s) lors de la fixation de la noradrénaline dans les fibres adrénergiques des Mammifères. Aussi peut on estimer que les différences observées entre les caractéres spectrofluorimétriques des catécholamines de G. convoluta et du Rat pourraient tenir plus à la forme de capture et de stockage des amines qu'à une différence profonde de leur nature chimique.

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Biogenic amine study of Glycera convoluta K. (annelida, polychaeta)

Summary The supra-oesophageal ganglion and ventral nerve cord of Glycera convoluta K. were studied with the fluorescence method of Falck. Many perikarya and fibers show a green and yellow fluorescence. The nature and metabolism of the observed biogenic amines were studied by biochemical methods. Epinephrine and 5-hydroxytryptamine were certainly present. Two other amines show chromatographic characters of dopamine and norepinephrine, but exhibit a slight difference in their fluorimetric spectra with these two catecholamines.Radiochemical methods have shown that the retention of norepinephrine and dopamine is longer in the ventral nerve cord of G. convoluta than in the vas deferens of the rat.The lipids, proteolipids and proteins of G. convoluta were different from those found in mammals as demonstrated by chromatographic analysis.Recent findings have shown the importance of the spatial arrangement of active groups in the molecule of transmitter for its retention by the protein matrix in the adrenergic fibers of mammals. The differences found by spectrofluorimetry and by measurements of the time of retention between catecholamines of rat and G. convoluta can be explained by differences in the nature of the protein matrix or by the spatial arrangement of active groups in the transmitter molecule.

Nos remerciements vont à Mme L. Desgroux-Lelièvre et à M.B. Lesbats, pour la collaboration qu'ils nous ont apportée dans la réalisation de ce travail, ainsi qu'au personnel de la Station Biologique de Roscoff qui nous a aimablement fourni les animaux utilisés.     

Phylogenetic position and physiology of Cerinosterus cyanescens

Partial 25S rRNA sequencing of Cerinosterus cyanescens showed it to be a close relative of Microstroma juglandis, a member of the basidiomycetous order Microstromatales. It is unrelated to the generic type species, C. luteoalba, which is a member of the order Dacrymycetales. The clinical occurrence of C. cyanescens is possibly explained by its thermotolerance and lipolytic activity. The species' nutritional profile is established. Growth on n-hexadecane is rapid; it grows well on typical plant constituents like gallic, tannic, vanillic, quinic and p-coumaric acids, but not on 3-hydroxybenzoic acid, phenol and hydroquinone. The failure to assimilate D-galactose, L-sorbose and ethylamine, the presence of urease and sensitivity to cycloheximide are diagnostic for the species.     

Visual and olfactory location of biotopes, prey patches, and individual prey by the ladybeetleChilocorus nigritus

Foraging behaviour of the predatorChilocorus nigritus (Fabricius) (Coleoptera: Coccinellidae) at the three spatial levels of biotope, prey patch and individual prey, was studied in the laboratory, and related to behaviour in the field. Vertically oriented parallel lines were more attractive than the same shapes in a horizontal position. A simulated horizon with a tree line was preferred to a simulated flat horizon. They were attracted to a tree image for the first 2 h of exposure, but were less attracted after longer exposure, possibly due to habituation. Leaf shape was recognised, and simple ovate leaves were preferred to compound bipinnate leaves and to squares. These responses were associated with biotope selection for feeding and aggregation at aestivation sites. The location of prey patches by adults involved prey odour but the location of such sites by larvae did not. Adults detected individual prey visually and olfactorily over short distances but physical contact with prey was required for detection by larvae. Location of individual prey and prey patches by adults and larvae was facilitated by alternation between intensive and extensive search. The differences in the ability of larvae and adults to locate prey, stem from the adults being the active locators of biotope and patch, whereas the comparatively immobile larvae depend on their parents’ ability for long-range location of prey. Two hypotheses concerning coccinellid foraging behaviour are proposed. Firstly, the duration of response to a visual cue is related to the distance over which such a cue may be perceived. It follows that habituation to closer range cues occurs more rapidly than to longer range cues. Secondly, visual cues used by adults at the different spatial levels of prey location, and the location of mates and aggregation sites, have the same or similar shape. These results also provide guidelines for orchard management to maximise the biocontrol value of this species.     

Morphology, anatomy, and taxonomic position of Pagameopsis (Rubiaceae-Rubioideae)

Morphological and anatomical features (including wood anatomy and pollen morphology) of the small neotropical genusPagameopsis (Rubiaceae-Rubioideae) are discussed and illustrated. The fused ovaries, fenestrate corolla tube, basally attached anther filaments, and absence of raphides are especially noteworthy.Pagameopsis is definitely not a member of the Psychotrieae because of significant differences in wood anatomy and gynoecial and fruit structure. A close affinity with Gaertnereae seems doubtful for similar reasons. The taxonomic affinities ofPagameopsis remain obscure. The genus shows similarities with several taxa of the Rubioideae, such as Coccocypseleae, Morindeae, Hedyotideae, andMetabolos. The absence of raphides, on the other hand, makes a position in the Rubioideae doubtful.