Adrenoreceptors, cyclic nucleotides, and the regulation of spleen cell antigen binding in urodele and anuran amphibians.
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Biomedical subjects
Publications and source records attributed to M Balls.
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The incubation of carrot protoplasts and cultured Xenopus cells in a protease solution has been shown to enhance their subsequent interkingdom fusion by a high pH/high Ca2+ method. The effects of Ca2+ concentration, pH, and temperature on the frequency of heterokaryon formation have also been studied. Potentially viable heterokaryons have been repeatedly produced at high frequencies (consistently greater than 10%), far exceeding those so far achieved in PEG-mediated fusion. Cell aggregates are readily dispersed after this method of fusion, permitting the accurate estimation of fusion frequencies.
Adults of two urodele amphibian species (Triturus cristatus carnifex and Cynops hongkongensis) and two anuran species (Rana temporaria and Xenopus laevis laevis) were immunized with a 25% suspension of sheep or horse erythrocytes. After eight or 14 days, splenic lymphocytes were removed, and their specific red cell-binding capacities tested by immunocytoadherence. Antigen-binding cells were classified as high-dose nonsecretory (S-) or secretory (S+), according to whether they bound a single layer or several layers or erythrocytes. The stimulation of both alpha and beta adrenoreceptors reduced the numbers of S+ rosettes formed by Triturus and Cynops lymphocytes, whereas a beta agonist increased and an alpha agonist decreased S+ rosette formation by Rana and Xenopus splenic lymphocytes. These effects were blocked by alpha and beta adrenoreceptor antagonists. Low-dose immunization of Xenopus with a 0.0025% suspension of sheep erythrocytes gave a minimal number of S+ rosettes two and eight days after immunization, and beta adrenoreceptor stimulation had no effect on antigen binding. These results are discussed in terms of the distribution of alpha and beta adrenoreceptors in amphibians and possible relationships between S+ and high-dose S- antigen-binding cells, and support the view that functional lymphocyte heterogeneity exists in these lower vertebrates.
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The renal corpuscle and segments of the renal tubule of the urodele amphibian Amphiuma means have been described and studied by light and electron microscopy as a background for investigations on the effects of hormones, drugs, toxins and carcinogens on cell structure and function in long-term organ cultures of kidney explants. Notable features of the A. means kidney include the number and variety of cytoplasmic inclusions in the cells of the proximal segment, the comparatively thick glomerular filter, and the presence of possible renin granules in the endothelial cells lining the afferent arteriole near its point of entry to the glomerular capsule.
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Isoprenaline, db-cAMP and ionophore Br-X537A induced glycogenolysis and glucose release from A. means liver organ cultures in the presence or absence of medium Ca2+. Ionophore A23187 had similar effects, but only in the presence of external Ca2+.
Secretion from the granular glands of Xenopus laevis skin was stimulated by alpha-adrenergic agonists, an effect which was blocked by alpha-adrenergic antagonists and inhibited by beta-adrenergic agonists, db-cAMP and diazoxide. The inhibition by isoprenaline and salbutamol, but not that by diazoxide, was blocked by a beta-adrenergic antagonist. It is concluded that the myoepithelial cells surrounding the secretory compartment of the granular glands bear alpha and beta adrenoceptors, and that the beta receptors comprise, or at least include beta2 receptors.
High concentrations (10 mM) of alanine, glycine, and glutamic acid in the culture medium had no effect on urea production in Amphiuma means liver in organ culture. Ammonia production was increased in media containing added alanine and glycine, but reduced in medium with added glutamic acid.
The sodium transport capacity across frog skin and toad bladder in vitro is enhanced when a tissue culture medium is used to mount the preparations, instead of the classic Ringer solution. The response to antidiuretic hormone used 24 h after isolating the tissues is also higher in preparations incubated in the same culture medium.
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Fragments of liver from the adult urodele Amphiuma means, the Congo eel, were maintained in organ culture for up to 70 days. The normal electrophoretic patterns of several enzymes were retained. The activities of ornithine transcarbamylase, arginase, glutamate oxalacetate transaminase and glutamate pyruvate transaminase, and urea production, glucose uptake and tissue glycogen content remained relatively constant throughout the culture period. Histological organization and hepatocyte ultrastructure were also retained. Liver fragments survived better in media based on MEM or BME than in medium based on Leibovitz L15. Since many aspects of tissue-specific structure and function are retained, long-term amphibian organ culture is well suited to studies on the control of hepatocyte function and on the effects of metabolites, hormones, drugs and toxins.