Zusammenfassung
In dieser Arbeit wird eine Methode beschrieben, mit der man mit höherer Auflösung Proteine in Gefrierschnitten vom Innenohr durch Immunofluoreszens lokalisieren kann. Die Gewebestückchen werden in Gelatine eingebettet und Schnitte von 0,1–1 μm Dicke an einem Kryoultramikrotom bei −100° C geschnitten. Die Schnitte werden mit Antikörpern gegen die Proteine des Cytoskelets Aktin und Tubulin markiert. Actin, das bisher nur in der Sinneszelle lokalisiert war, wird nun auch in den Stützzellen gefunden. Tubulin wird in den Stützzellen und in den äußeren Spiralnervenfasern gefunden.
Summary
The present work describes a high resolution technique for locating proteins in frozen sections of the inner ear by immunofluorescence. Dissected organs are encapsulated in gelatin, and sections 0.1–1 μm thick are cut at −100°C in a cryoultramicrotome. These are labelled with antibodies against two cytoskeletal proteins, actin and tubulin. Actin, which had previously only been described in the sensory cells, is found in the supporting cells as well. Tubulin is identified in the supporting cells and in outer spiral nerve fibres.
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References
Baumgartner B, Tokuyasu KT, Chrispels (1978) Localization of vicilin peptidohydrolase in the cotyledons of bean seedlings by immunofluorescence microscopy. J Cell Biol 79: 10–19
Bernhard W, Nancy HT (1964) Coupes á congélation ultrafines de tissus inclus dans la gélatine. J Micros (Paris) 3: 579
Coons AH, Kaplan MH (1950) Localization of antigens in tissue cells. II. Improvement in a method for the detection of antigens by means of fluorescent antibody. J Exp Med 91: 1
Engström H, Ades HW, Andersson A (1966) Structural pattern of the organ of Corti. Almqvist and Wiksell, Stockholm
Flock Å, Cheung HC, Flock B, Utter G (1981). Three sets of actin filaments in sensory cells of the inner ear. Identification and functional orientation determined by gel electrophoresis, immunofluorescence, and electron microscopy. J Neurocytol 10: 133–147
Flock Å, Bretscher A, Weber K (1981) Immunohistochemical localization of several cytoskeletal proteins in inner ear sensory and supporting cells. Hearing Res (submitted)
Macartney JC, Comis SD, Pickles JO (1980) Is myosin in the cochlea a basis for active motility? Nature 288: 481–492
Schätzte W (1971) Histochemie des Innenohres. Urban u. Schwarzenberg, München Berlin Wien
Sevéus LL (1979) The subcellular distribution of electrolytes. A methodological study using cryoultramicrotomy and X-ray microanalysis. LKB, Stockholm
Tokuyasu KT (1973) A technique for ultracryotomy of cell suspensions and tissues. J Cell Biol 57: 551–565
Tokuyasu KT, Singer SJ (1976) Improved procedures for immunoferritin labelling of ultrathin frozen sections. J Cell Biol 71: 894–906
Weber K, Rathke PC, Osborn M and Franke WW (1976) Distribution of actin and tubulin in cells and in glycerinated cell models after treatment with cytochalasin B. Exp Cell Res 102: 285–297
Weber K, Wehland J, Herzog W (1976) Griesofulvin interacts with microtubules both in vivo and in vitro. J Med Biol 102: 817–829
Zenner HP (1981) Cytoskeletal and muscle-like elements in cochlear hair cells (in press)
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Supported by grants from the Swedish Medical Research Council (no. 04x-02461), the Ragnar and Torsten Söderberg Foundation, and the Foundation Tysta Skolan
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Flock, Å., Hoppe, Y. & Wei, X. Immunofluorescence localization of proteins in semithin 0.2–1 μm frozen sections of the ear. Arch Otorhinolaryngol 233, 55–66 (1981). https://doi.org/10.1007/BF00464275
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DOI: https://doi.org/10.1007/BF00464275