Glucose Transporter (GLUT-4) Is Targeted to Secretory Granules in Rat Atrial Cardiomyocytes

The insulin-responsive glucose transporter GLUT-4 is found in muscle and fat cells in the transGolgi reticulum (TGR) and in an intracellular tubulovesicular compartment, from where it undergoes insulindependent movement to the cell surface. To examine the relationship between these GLUT-4–containing compartments and the regulated secretory pathway we have localized GLUT-4 in atrial cardiomyocytes. This cell type secretes an antihypertensive hormone, referred to as the atrial natriuretic factor (ANF), in response to elevated blood pressure. We show that GLUT-4 is targeted in the atrial cell to the TGR and a tubulo-vesicular compartment, which is morphologically and functionally indistinguishable from the intracellular GLUT-4 compartment found in other types of myocytes and in fat cells, and in addition to the ANF secretory granules. Forming ANF granules are present throughout all Golgi cisternae but only become GLUT4 positive in the TGR. The inability of cyclohexamide treatment to effect the TGR localization of GLUT-4 indicates that GLUT-4 enters the ANF secretory granules at the TGR via the recycling pathway and not via the biosynthetic pathway. These data suggest that a large proportion of GLUT-4 must recycle via the TGR in insulin-sensitive cells. It will be important to determine if this is the pathway by which the insulin-regulatable tubulo-vesicular compartment is formed.     

Human leukemic K562 cells: differential effects of 5-azacytidine on DNA methylation of epsilon-, gamma-globin and 7SL RNA genes

5 Azacytidine ribonucleoside (5 Aza CR), greatly enhances erythroid differentiation of the K562(h) cell line, with a sharp increase of embryonic and fetal globin gene expression. This phenomenon is correlated with the undermethylation of gamma-globin but not of epsilon-globin, as the epsilon-globin gene is already extensively undermethylated before 5AzaCR induction. By contrast no variations in both DNA methylation and expression are observed in 7SL RNA genes.     

Besondere Haarstrukturen der Soricidae (Mammalia,Insectivora) und ihre taxonomische Deutung

Zusammenfassung Die vorliegende Untersuchung befaßt sich mit besonderen Haarfeinstrukturen der Soricidae, wobei geklärt werden soll, ob dem H-förmigen Haarquerschnitt-Profil eine taxonomische Bedeutung zukommt. Wir überprüften deshalb die betreffenden Haarstrukturen mit Hilfe des REM in 8 Gattungen.Das besondere Haarprofil, das auf das Terminalsegment der Grannenhaare beschränkt ist, findet sich bei folgenden Gattungen:Sorex, Neomys, Blarina undCryptotis, alles Vertreter der Subfamilie Soricinae. Sämtliche untersuchten Vertreter der Subfamilie Crocidurinae, d.h.Crocidura, Praesorex, Suncus undSylvisorex weisen ein einfaches Haarprofil auf.Das H-Profil wird als Synapomorphie der Soricinae angesehen und charakterisiert diese als monophyletische Gruppe. Die haarmorphologischen Kriterien ergänzen somit die osteologischen Kriterien von Repenning (1967) und sprechen für die Beibehaltung der von vielen Autoren abgelehnten Subfamilien.

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Special hair structures in Soricidae (Mammalia, Insectivora) and their taxonomic interpretation

Summary The following study should clear up the structures of the H-shaped profile found in the hairs of some shrews and show if it has a taxonomic signification. Therefore we studied the concerned hair structures by scanning electron microscopy in 8 genera.The special hair-shape, which is confined to the terminal segment of guard hairs, is found in the species of the following genera:Sorex, Neomys, Blarina andCryptotis, all members of the subfamily Soricinae. All the examined members of the subfamily Crocidurinae, i.e.Crocidura, Praesorex, Suncus andSylvisorex show a simple hair shape.The H-shaped hair characterizes the Soricinae as a monophyletic unity. Yet, the morphological criteria of hair complete the osteological criteria of Repenning (1967) an plead for the validitiy of the often refuted subfamilies.

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Mit Unterstützung des Schweizerischen Nationalfonds zur Förderung der wissenschaftlichen Forschung (Nr. 3.515.71, 3.821.72, 3.413-0.74)

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Unser Dank gilt Herrn Prof. N. Schönenberger, Herrn Prof.R. Krstic und Frau C. Regamey, die uns bei der histologischen Präparation behilflich waren, insbesondere auch Herrn Dr. T. Jalanti, der uns in die REM-Technik einführte. Kostbares Material wurde uns von Frau A. Geraets (Bonn), Herrn Prof. U. Rahm (Basel) und Herrn Dr. V. Aellen (Genf) zur Verfügung gestellt; auch ihnen sei hierfür herzlich gedankt.     

Evaluation Of Eight Fluorochrome Combinations For Simultaneous Dna-Protein Flow Analyses

Eight fluorescent dye combinations for simultaneous DNA-protein staining have been evaluated spectroscopically and flow microfluoromctrically: propidium iodide (PI) with fluorescein-isothiocyanate (FITC), fluorescamine (FC), and dansylchloride (DANS); diamidinophenylindole (DAPI) with sulphorhodamin (SR101), tetramethylrhodamin isothiocyanate (TRITC), and nitroben-zodiazole (NBD); acriflavine (AF) with stilbene isothiocyanate sulphonic acid (SITS), and DAPI. Three different experimental tumor cell lines have been employed in the investigations. Simultaneous DNA-protein analyses have been carried out with the newly developed HEIFAS instrument. Spectroscopically two groups of dyes were distinguishable according to their excitation maximum below 400 nm and above 450 nm respectively. DANS and NBD were found to be unsatisfactory with respect to their protein distributions obtained by flow analysis. The remaining stains involved in the dye combinations revealed comparable flow distributions of the cellular DNA and protein content. With respect to preparation time and number of centrifugal steps involved in the staining protocols, and in connection with the stability of the dye used, the DAPI-SR101 method proved to be fastest and easiest With this combination DNA and protein flow analysis can be performed simultaneously within 30 min.     

Hydrogen-ion binding by tobacco-mosaic-virus protein polymers.

Hydrogen ion titration curves of tobacco mosaic virus protein have been measured in various conditions of protein concentration, temperature, ionic strength, and rate of pH change. The polymers present at each stage are deduced from turbidity and sedimentation data, plus published information. A simple semi-quantitative analysis of the curves is given, and the pK values of the two abnormal carboxylates in single helix are estimated as 6.4 and about 7.0. Disks, and some faster-forming unknown polymers in the same size range, have been abnormal carboxylate with pK 6.9. These results are most easily interpreted in terms of electrostatic interactions between carboxylates, probably at the axial ends of the protein subunits.     

Small-angle X-ray-scattering investigation and structural-model study of the fatty-acid synthetase from pig liver (author's transl)

The structure of the fatty acid synthetase from pig liver was studied on models based upon structural and functional properties selected from pertinent results available from numerous investigations carried out with fatty acid synthetases from this and other sources. When comparing small-angle X-ray-scattering curves calculated with these models and curves obtained from small-angle X-ray-scattering experiments carried out with the pig-liver enzyme, we tried to select a model which would lead to an acceptable correlation between the calculated and the experimental curves and at the same time fulfil the known structural and functional requirements. The comparison of the curves was started with a model of low complexity. The observed discrepancy, together with arguments from the structural and the functional properties, helped decide which is the next most reasonable model to be considered. This procedure was repeated for five models of increasing complexity. In the model which led to the best fit the multienzyme complex is composed of two halves in an assymetric conformation including hollow spaces. This highly anisotropic model would imply that the two halves change their conformation each time a synthetic cycle is completed and that the growing fatty acid is handed over from one half to the other.