Alpha-melanotropin in the frog brain: regional distribution, quantification and subcellular distribution

The distribution of alpha-MSH containing neurons was studied by immunofluorescence in the brain of the frog Rana ridibunda. Most immunoreactive cell bodies were found in the ventral hypothalamic area. A rich network of fluorescent fibers was observed in the ventral infundibular region, coursing towards the preoptic area and the ventral telencephalon. Some fibers, directed backwards, project into median eminence. By means of a specific radioimmunoassay, the concentrations of alpha-MSH immunoreactive material has been determined in 10 different regions of the brain. The highest concentrations were observed in the infundibular and the preoptic regions. Using the immunogold technique, electron microscopy showed that immunostaining was restricted to 70-100 nm dense core vesicles in positive cell bodies and fibers. These results suggest that, in addition to well known hormonal (melanotropic) activity, alpha-MSH could play the role of a neurotransmitter in the frog brain.     

Receptor-mediated particle uptake by liver macrophages. The galactose-particle receptor mediates uptake via coated and also non-coated structures

The endocytosis pathways of particles with terminal beta-D-galactosyl groups were studied in isolated rat Kupffer cells by electron microscopy. Colloidal gold particles of sizes 5, 17 and 50 nm were coated with asialofetuin (ASF) and isolated liver macrophages were allowed to bind (at 4 degrees C) or take up (at 37 degrees C) these ligands. Particles of all three sizes were bound via the galactose-particle receptor as shown by carbohydrate inhibition experiments and were ingested effectively. But, whereas ASF-gold particles of sizes 5 and 17 nm are taken up via the coated pit/coated vesicle pathway, the 50 nm particles are not. These enter the cell via non-coated endocytic vacuoles. All three particle sizes are transported to the same lysosomal compartment. These observations demonstrate that at least in macrophages one receptor is capable to mediate endocytosis via two different pathways depending on ligand size and/or valency.     

Production and purification of monoclonal antibodies to Pisum and Avena phytochrome

At least nine monoclonal antibodies against phytochrome from Pisum sativum L. and 20 against phytochrome from Avena sativa L. have been obtained from mouse hybridomas that were produced by fusion of spleen cells with SP 2/O-Ag14 myeloma cells. Hybridomas were selected and cloned in a single step by plating on a semisolid methylcellulose medium. Eight antibodies to Pisum and one to Avena phytochrome were immunopurified from hybridoma medium or ascitic fluid. When necessary, secreted antibodies were verified to be against phytochrome by demonstrating to be against phytochrome by demonstrating immunoadsorption of phytochrome, detected as loss of photoactivity and-or by appearance of the approx. 120,000-dalton phytochrome band upon sodium dodecyl sulfate polyacrylamide gel electrophoresis.     

Ultrastructure des cellules germinales au cours du développement embryonnaire du Lézard vivipare (Lacerta vivipara Jacquin)

Résumé Les gonocytes primaires sont relativement pauvres en polysomes et l'ergastoplasme granulaire est très réduit. Le reticulum endoplasmique de type lisse se développe au cours du développement embryonnaire. L'appareil de Golgi est bien représenté et localisé au niveau de la calotte juxtanucléaire mitochondriale. Les liposomes cytoplasmiques sont très nombreux.Cette étude précise la structure du nucléole «annulaire» et de la «masse paranucléolaire» observés en microscopic photonique. Des modifications nucléolaires sont constatées au cours du développement embryonnaire. Certains aspects ultrastructuraux sont vraisemblablement en rapport avec le déplacement autonome des gonocytes. La signification physiologique du nucléole «annulaire» et de la «masse paranucléolaire» est envisagée.

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Ultrastructural study on the primordial germ cells during embryonic development of Lacerta vivipara Jacquin

Summary The primordial germ cells of Lacerta vivipara have relatively few free polysomes and little ergastoplasm. Smooth endoplasmic reticulum increases during embryonic development. The Golgi apparatus is well developed and lies close to the mitochondrial juxtanuclear cap. This study shows the ultrastructure of the ring-shaped nucleoli and the masse paranucléolaire. Modifications in nucleolar structure are observed during embryonic development. Some ultrastructural features are probably related to the ameboid movement of the primordial germ cells. The physiological meaning of the ring-shaped nucleoli and the masse paranucléolaire is considered.

Avec la collaboration technique de Mme. M. Hubert.     

Die auswirkungen der körpergrösse von Aphelinus semiflavus howard (HYM., Aphelinidae) auf einige Organe und ihre leistungsfähigkeit

Zusammenfassung Die Körpergröße von Aphelinus semillavus How. variiert in Abhängigkeit von Große und Qualität des Wirtes. Tiere aus dem gesamten Variationsbereich wurden auf größenabhängige Proportionsverschiebungen untersucht.Relative Länge und Breite des Kopfes (bezogen auf die Körperlange) nehmen mit zunehmender Körperlange ab. Die relative Lange des Abdomens wächst etwa um den gleichen Betrag:Es besteht eine negative Korrelation zwischen der relativen Länge des letzten Fühlergliedes, des Trägers wichtiger Sinnesorgane, und der Körperlange. Seine Oberfläche steht in einem etwa konstanten Verhältnis zur Körperlänge. Die Anzahl der Porenplatten nimmt mit der Körperlänge nur wenig, ihre Länge deutlicher zu.Im Komplexauge wächst die Anzahl der Ommatidien etwa proportional zur Korperlange, während seine gesamte Fläche zwar linear, aber ïiberproportional ansteigt, was durch zunehmende Große der einzelnen Facetten erreicht wird.Die Länge der Vorderfliigel ist bei Männchen und Weibchen von mehr als 1 mm Länge etwa der Körperlange proportional. Unterhalb 1 mm nimmt die relative Flügellänge mit der Körperlange ab, bis die Flügel nur noch funktionslose Rudimente bilden. Die Relation Flügel-fläche:Körpergewicht ist bei mittelgroßen Tieren am günstigsten.Jedes Ovar hat stets drei Ovariolen, unabhängig von der Körperlänge. Große Weibchen legen nur etwas größere, aber bedeutend mehr Eier als kleine. Die Anzahl der (am 7.–10. Lebenstag) täglich abgelegten Eier steigt etwa linear mit dem Logarithmus des Körpergewichts an.Die Eiablage wird während des ganzen Lebens (im Labor etwa 40 Tage) fortgesetzt, ist aber zum. Ende hin etwas geringer. Relativ zum Körper-gewicht produzieren mittelgroße Weibchen die größte Eimasse. Die Anzahl der zum Nahrungsgewinn ausgesaugten Läuse steigt mit dem Körpergewicht.Zwischen der relativen Länge des Legestachels und der Körperlange besteht eine fast lineare negative Korrelation.Die gröBenabhangigen Proportionsverschiebungen werden als Auswirkungen unterschiedlicher Wachstumstendenzen der einzelnen Organe gedeutet, die ihrerseits genetisch fixiert sind und den biologischen Sinn haben, die Lebensf ähigkeit modifikatorisch kleiner Exemplare zu sichern und damit die Ausnutzung entsprechend kleiner Wirte zu ermöglichen.Die Bedeutung der Proportionsverschiebungen für die Leistung der ganzen Population wird diskutiert. Sic ist abhängig von der Häufigkeits-verteilung der verschiedenen Körpergrößen und these wiederum von der Art-Zusammensetzung und Ernährung der Wirtsfauna.

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Summary The body size of Aphelinus semiflavus How. varies with the size and the quality of the hosts. Specimens from the whole range of variation were investigated for differences between organ proportions, which depended on size.The relative length and width of the head (based on the body length) are reduced with increasing body length. The relative length of the abdomen increases approximately by the same amount.There is a negative correlation between the relative length of the last joint of the antennae and the body length. Its surface shows a nearly constant proportion to the body length. The length of the pore plates increases with the body length more distinctly than their numbers.Within the compound eye, the number of ommatidia increases almost proportionally to the body length; the whole area of the eye increases more than proportionally because the size of the single facets also becomes greater.When the body length is greater than 1 mm, the length of the forewings is nearly proportional to the body length. At less than 1 mm, the relative length of the fore-wings is reduced with declining body length until, finally, only rudiments without any function remain. The ratio between wing area and body weight is most favourable at medium body size.Three ovarioles, independent of the body size, are present in each ovary. Big females produce considerably more eggs than small ones. The size of their eggs is only slightly enlarged. The daily egg production (at the 7^th–10^th day of life) increases nearly linearly with the logarithm of body weight. Oviposition is continued throughout the life-time of the insect (ca. 40 days in the laboratory) but it is somewhat reduced at the end. Relative to their body weight, females of medium size produce the biggest egg masses. The number of aphids utilized by the parasite for nourishment increases with the body weight of the female.A negative correlation exists between the relative length of the extended ovipositor and the body length.The proportion differences of the organs, dependent on size, are interpreted to be consequences of different growth tendencies which are genetically fixed. Apparently, the biological role of such different growth tendencies is to ensure the viability of phenotypically small specimens and in this way to make the utilisation of small hosts possible.The importance of the differences of body proportions for the effect of the whole population is discussed. It depends on the frequency distribution of the different body lengths which itself is the consequence of the composition of host species and the nutriation of the hosts.

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Herrn Prof. Dr. Hans Braun, emer. Direktor des Instituts für Pfanzenkrankheiten an der Universität Bonn, zu seinem 70. Geburts'ag am 2. Februar 1966 gewidmet.     

Direct effects of serotonin and ketanserin on the functional morphology of embryonic chick skin in vitro

Summary Different organotypical culture methods are used to test the direct effects of serotonin and ketanserin, a S₂, α₁, and H₁ receptor antagonist in vascular tissue, on fibroblasts and epidermal cells of embryonic chick skin in vitro. From light microscopic and electron microscopic analyses, we learn that serotonin enhances keratinization and differentiation, whereas ketanserin reduces differentiation in comparison to the control cultures. Incorporation data of fragments cultured with [³H]thymidine show that ketanserin, within a dose range from 0.05 to 5 μg/ml, stimulates proliferation. Serotonin at a concentration of 10 μg/ml slightly slows down proliferation, whereas lower doses of 0.1 and 1 μg/ml result in tritium activities that do not differ from control cultures. This investigation was financially supported by the National Fund of Scientific Research, Belgium, 3.0022.87.     

Rabbit distal colon epithelium: III. Ca2+-activated K+ channels in basolateral plasma membrane vesicles of surface and crypt cells

Summary In the mammalian distal colon, the surface epithelium is responsible for electrolyte absorption, while the crypts are the site of secretion. This study examines the properties of electrical potential-driven⁸⁶Rb⁺ fluxes through K⁺ channels in basolateral membrane vesicles of surface and crypt cells of the rabbit distal colon epithelium. We show that Ba²⁺-sensitive, Ca²⁺-activated K⁺ channels are present in both surface and crypt cell derived vesicles with half-maximal activation at 5×10^–7 m free Ca²⁺. This suggests an important role of cytoplasmic Ca²⁺ in the regulation of the bidirectional ion fluxes in the colon epithelium.The properties of K⁺ channels in the surface cell membrane fraction differ from those of the channels in the crypt cell derived membranes. The peptide toxin apamin inhibits Ca²⁺-activated K⁺ channels exclusively in surface cell vesicles, while charybdotoxin inhibits predominantely in the crypt cell membrane fraction. Titrations with H⁺ and tetraethylammonium show that both high-and low-sensitive⁸⁶Rb⁺ flux components are present in surface cell vesicles, while the high-sensitive component is absent in the crypt cell membrane fraction. The Ba²⁺-sensitive, Ca²⁺-activated K⁺ channels can be solubilized in CHAPS and reconstituted into phospholipid vesicles. This is an essential step for further characterization of channel properties and for identification of the channel proteins in purification procedures.