Effect of vasoactive amines on Weibel-Palade bodies in capillary endothelial cells

The presence and distribution of Weibel-Palade bodies in stomach and colonic mucosal microvessels after the administration of vasoactive amines (serotonin and histamine), the serotonin depletor reserpine, and the von Willebrand factor secretagogue thrombin, was studied by transmission electron microscopy. These agents elevated the number of Weibel-Palade bodies in all microvascular endothelial cells and especially in capillaries. It is concluded that vasoactive amines enhance the synthesis and secretion of large von Willebrand protein multimers by endothelial cells.     

Neuromodulation by serotonin and octopamine in the honeybee: behaviour,neuroanatomy and electrophysiology

The biogenic amines serotonin (5HT) and octopamine (OA) exist in the bee and can modulate neuronal activity and behaviour. 5HT-like and OA-like immunoreactivities can be found in most neuropils of the brain. Binding sites for the two amines are also present in most brain neuropils. The highest density of binding sites for [³H]serotonin and [³H]octopamine was found in the mushroom bodies. In some brain areas, especially the mushroom bodies, mismatches exist between binding sites and immunoreactivities, suggesting that the two amines also bind to neuropils which are not directly innervated by 5HT-like or OA-like immunoreactive neurons. The action of the two amines on behaviour in the bee is antagonistic. In the antennal pathway, proboscis and antennal responses to olfactory and gustatory stimuli are enhanced by OA and reduced by 5HT. In olfactory conditioning experiments, storage and retrieval of the learned signal can be enhanced by OA and reduced by 5HT. The specificity of these effects depends on the site of amine application in the neuropil. In the visual system the direction specificity of the visual antennal response is enhanced by OA and reduced by 5HT after topical application or injection into the lobula, the third optic ganglion. Correlates for the behavioural modulation can be found in higher-order visual interneurons. While OA application can mimic the stimulation of the bee with sugar water, the behavioural conditions leading to the release of 5HT are not yet known.     

Shear-dependence of endothelial functions

Endothelial cells are subjected to shear forces which influence important cell functions. Shear stress induces cell elongation and formation of stress fibers, increases permeability, pinocytosis and lipoprotein internalization, is involved in the formation of atherosclerotic lesions, increases the production of tissue plasminogen activator, and enhances von Willebrand factor release and hence platelet aggregation. It decreases adherence of erythrocytes and leukocytes, and increases the release of prostacyclin, endothelium derived relaxing factor, histamine and other compounds, but decreases erythropoietin secretion. The mechanism of signal transduction to the endothelial cell is not known exactly; shear-sensitive ion channels seem to be involved. It is concluded that a better understanding of shear-dependent endothelial functions will influence pathophysiologic concepts and therapeutic interventions.     

Macrophage origin of platelet activating factor]

Platelet-activating factor (P.A.F.) is a mediator of anaphylaxis released from human and Rabbit basophils which causes aggregation of platelets and release of their vasoactive amines. We have induced the release of P.A.F. from Rat peritoneal cells (P.C.) with ionophore A 23187. After fractionation of P.C. on 5-15% Ficoll gradients, P.A.F. was obtained from macrophage-rich but not from mastocyte-rich fractions and from adherent cells but not from non adherent cells. These data suggest an important new function for the macrophage: aggregation of platelets and release of their vasoactive amines and others mediators of inflammation.     

Insights from von Willebrand disease animal models

von Willebrand disease is a genetic bleeding disorder that arises from abnormalities in von Willebrand factor, an adhesive glycoprotein involved in both primary hemostasis and coagulation. It is the most common inherited bleeding disorder in humans, and over the years several animal species have also been described as suffering from this disease whether through a spontaneous mutation (pigs, dogs) or a genetically engineered one (mouse). These different animal models are extremely useful in exploring the characteristics of von Willebrand disease and in testing new treatments. This review provides an update of the various von Willebrand disease models and the contribution that these models can make to a better understanding of human von Willebrand disease.     

Serotonin is localized in endothelial cells of coronary arteries and released during hypoxia: A possible new mechanism for hypoxia-induced vasodilatation of the rat heart

Summary In this report we demonstrate the immunocytochemical localization of serotonin in endothelial cells of rat coronary vessels and a significant increase in the release of serotonin into the perfusate of Langendorff rat heart preparations during hypoxia. It is suggested that serotonin, localized in endothelial cells, is released during hypoxia and could provide part of a pathophysiological mechanism for vasodilatation to protect the heart from damage due to hypoxia.This research was supported by the British Heart Foundation and the excellent technical assistance of Jon Bokor is gratefully acknowledged.     

Distribution patterns of mammalian-like peptide immunoreactive cells in the midgut ofAeshna cyanea (Insecta,Odonata)

Summary The distribution of cells reacting with antisera to cholecystokinin, substance P, gonadoliberin, methionine-enkephalin, and vasoactive intestinal peptide, demonstrated by the indirect immunoperoxidase method, was studied along the entire midgut of an insect,Aeshna cyanea. For each antiserum, the number of reacting cells increased from the middle part to the end of the midgut. Only a few cells reacted to somatoliberin, leucin-enkephalin and somatostatin antisera. In the connective sheath surrounding the midgut epithelium, nerve fibers were stained by antisera to serotonin, somatostatin, cholecystokinin, vasoactive intestinal peptide and methionine-enkephalin.     

Serotonin is localized in endothelial cells of coronary arteries and released during hypoxia: a possible new mechanism for hypoxia-induced vasodilatation of the rat heart

In this report we demonstrate the immunocytochemical localization of serotonin in endothelial cells of rat coronary vessels and a significant increase in the release of serotonin into the perfusate of Langendorff rat heart preparations during hypoxia. It is suggested that serotonin, localized in endothelial cells, is released during hypoxia and could provide part of a pathophysiological mechanism for vasodilatation to protect the heart from damage due to hypoxia.     

The diet and gut microflora influence the distribution of enteroendocrine cells in the rat intestine

Several functions of the gut are locally influenced by peptides and biogenic amines released from enteroendocrine cells. The aim of the present study was to assess whether the luminal stimulus of diet or microbial flora or diet-microbial interactions have an influence on the distribution of enteroendocrine cells along the crypt-surface axes of the small and large intestine. The effects of diet and indigenous flora were investigated by comparing the numbers of argyrophil and serotonin immunoreactive cells in the jejunum and colon of germ free and conventional rats fed either a purified diet containing fine ingredients or a commercial diet containing crude fibre of cereal origin. The effects of human flora were analysed in germ-free rats inoculated with human faecal organisms. 1. Feeding the commercial diet reduced the number of argyrophil endocrine cells in the jejunum and serotonin immunoreactive cells in the colon of gern-free animals but increased the serotonin immunoreactive cells in the colon of conventional animals. 2. The rat flora increased the serotonin immunoreactive cells in the colon of animals fed a commercial diet and decreased in those fed a purified diet. 3. Inculation of human flora increased the numbers of serotonin immunoreactive cells both in the jejunum and colon. The results provide evidence that the dietary changes and diet-microbial interactions can affect the regional number of enteroendocrine cells.     

Untersuchungen zur Klärung der Funktion des Reissnerschen Fadens. In-vitro-Bindung von Noradrenalin,Adrenalin und Serotonin

Summary The glycoprotein of Reissner's fibre is able to bind norepinephrine, epinephrine and serotonin. Bound amines can be demonstrated at the RF, even after washing with artificial cerebrospinal fluid for some, hours. It is discussed whether it is a function of the RF to remove substances, for instance biogenic amines, from the cerebrospinal fluid.     

Defective ristocetin and bovine factor VIII-induced platelet aggregation in normal rats

Riassunto Le piastrine di 3 diversi ceppi di ratti, contrariamente alle piastrine umane e di cavia, non sono aggregate (sia in vitro che in vivo) nè dalla ristocetine nè dal fattore von Willebrand bovino, porcino o umano, quest'ultimo reso aggregante previo trattamento enzimatico. Tale insensibilità, di cui si è potuta escludere l'origine plasmatica, potrebbe derivare dall'assenza, sulle piastrine di ratto, di un recettore per il fattore von Willebrand (acido sialico?); potrebbe essere cosi spiegata anche la mancata risposta delle piastrine alla ristocetina. Questi risultati suggeriscono che le piastrine di ratto possono essere un utile modello sperimentale per lo studio della sindrome di Bernard-Soulier nell'uomo.

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This work was supported by Contract No. 73-00400-04 of the C.N.R., Roma, Italy. The skilful technical assistance of MissAnnalisa Cavenaghi is appreciated.     

Serotoninergic innervation of the paraventricular organ in chick embryo and chick; histofluorescence and autoradiography study optic and electron microscopy]

Fluorescence microscopy (Falck and Hillarp technique) of 11th day old chick embryo shows aminergic neurons and processes in the paraventricular organ area. Administration of tritiated amines permits us to observe uptake and storage of serotonin only as early as the 10th day; this suggests a very early hypothalamic serotoninergic nervous system differentiation.     

Extraneuronal serotonin accumulation in peripheral arteries of the rat

Accumulations of serotonin (5-HT) and norepinephrine (NE) were compared in control and 6-hydroxydopamine (6-OHDA) pretreated rat aorta, mesenteric and tail arteries. The distribution of these amines was corrected by subtracting tissue uptake of tritiated sorbitol in the extracellular space. 5-HT greatly accumulated both in control and 6-OHDA pretreated arteries. In contrast, NE accumulation in mesenteric and tail arteries was substantially decreased after 6-OHDA treatment. In the aorta 6-OHDA pretreatment did not affect the accumulation of both amines. These findings suggest that 5-HT accumulation in these arteries is mainly extraneuronal, and NE mainly neuronal. Since the accumulation of 5-HT in the aorta was not influenced by pretreatment with 10 microM NE, the extraneuronal uptake mechanisms for 5-HT and NE appear to be different.     

ADAMTS-2 functions as anti-angiogenic and anti-tumoral molecule independently of its catalytic activity

ADAMTS-2 is a metalloproteinase that plays a key role in the processing of fibrillar procollagen precursors into mature collagen molecules by excising the amino-propeptide. We demonstrate that recombinant ADAMTS-2 is also able to reduce proliferation of endothelial cells, and to induce their retraction and detachment from the substrate resulting in apoptosis. Dephosphorylation of Erk1/2 and MLC largely precedes the ADAMTS-2 induced morphological alterations. In 3-D culture models, ADAMTS-2 strongly reduced branching of capillary-like structures formed by endothelial cells and their long-term maintenance and inhibited vessels formation in embryoid bodies (EB). Growth and vascularization of tumors formed in nude mice by HEK 293-EBNA cells expressing ADAMTS-2 were drastically reduced. A similar anti-tumoral activity was observed when using cells expressing recombinant deleted forms of ADAMTS-2, including catalytically inactive enzyme. Nucleolin, a nuclear protein also found to be associated with the cell membrane, was identified as a potential receptor mediating the antiangiogenic properties of ADAMTS-2.     

Der Einfluss von Calcium auf die Brenzcatechinaminfreisetzung

Summary (1) Experiments on isolated perfused suprarenals of cattle have shown that the acetylcholine-induced release of catechol amines, but not that of phenylethylamine, is dependent on calcium, since a perfusion with calcium-free Tyrode's solution abolishes the action of acetylcholine and not that of phenylethylamine. (2) Inincubation experiments with isolated chromaffin granules, calcium produces a dose-dependent, significant release of catechol amines even in physiological concentrations (2.5 mM). (3) Acetylcholine does not release catechol amines from isolated granules, either in the presence or in the absence of calcium.

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Forschungs-Stipendiat der A. von Humboldt-Stiftung.

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Ausgeführt mit Unterstützung der Deutschen Forschungsgemeinschaft.     

Thrombospondins: from structure to therapeutics

Thrombospondin-1 (TSP1) is a multi-domain, multi-functional glycoprotein synthesized by many cells. Matricellular TSP1 modulates cell adhesion and proliferation. TSP1 is involved in angiogenesis, inflammation, wound healing and cancer. As a major platelet protein, for a long time it was postulated to control hemostasis via platelet aggregate stabilization. However, these in vitro findings have been questioned in the absence of corroborating clinical data and of obvious hemostatic defects in TSP1 gene-deficient mice.Yet, the past few years have provided indices to implicate TSP1 in hemostasis. In clinical studies, a correlation exists between a welldefined TSP1 polymorphism and a significant risk of myocardial infarction.At the same time, recent in vivo animal model data imply TSP1 in the multimer size control of von Willebrand factor, in smooth muscle cell regulation and in vascular perfusion. These findings shed new light on the role of TSP1 in hemostasis and prothrombotic vascular pathologies. (Part of a Multi-author Review).     

Radioautographic identification of central monoaminergic neurons by local micro-instillation of tritiated serotonin or noradrenaline in cats]

Monoaminergic neurons in nuclei raphe dorsalis and locus coeruleus of the Cat may be visualized by radioautography after local micro-instillation of tritiated serotonin and noradrenaline. The concomitant administration of the appropriate tracer with the other biogenic amine in non radioactive form permits a specific identification of serotoninergic and catecholaminergic nerve cell bodies. A small contingent of presumptive serotoninergic neurons is thus demonstrated in the region of the locus coeruleus.     

La barrière capillaire enzymatique pour la DOPA au niveau de quelques noyaux du tronc cérébral du rat

Summary An enzymatic barrier for Dopa exists at the level of the cerebral capillaries which changes Dopa into Dopamine by means of endothelial decarboxylase. It is abolished by a decarboxylase inhibitor. Its intensity is weaker in structures containing Dopamine (Striatum, Locus Niger). Neurones of the Raphé containing serotonin can be charged with catecholamines. There are close contacts between neurones and capillaries.     

Monoamine-containing cells in atrioventricular valves of the opossum heart

Summary Using a specific technique for biogenic amines, similar cells to those described as small intense fluorescent (SIF) cells were identified in the atrioventricular valves of the opossum heart. It is suggested that these cells, under neural control, may secrete amines.     

Monoamines in brain and urine of rats with hereditary hypothalamic diabetes insipidus

Summary Monoamine levels in brain and urine of homozygous and heterozygous diabetes insipidus (DI) rats (Brattleboro strain) were assessed. Homozygous DI rats had a higher whole brain content of serotonin than their heterozygous littermates. However, when corrected for differences in brain weight, homozygous DI also appeared to have higher brain concentrations of noradrenaline, tyrosine and GABA. The total 24 h excretion of all amines and their precursors was greater in the homozygous than in the heterozygous rats.The authors thankHerman Müller andIneke van de Veerdonk for their assistance with part of this study.