Use of a biotinylated probe and in situ hybridization for light and electron microscopic localization of P 0 mRNA in myelin-forming Schwann cells |
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Authors: | H. F. de Webster L. Lamperth J. T. Favilla G. Lemke D. Tesin L. Manuelidis |
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Affiliation: | (1) Laboratory of Experimental Neuropathology, NINCDS, National Institutes of Health, Bldg 36, 20892 Bethesda, MD, USA;(2) Section of Neuropathology, Yale University School of Medicine, 06510 New Haven, CT, USA;(3) Molecular Neurobiology Laboratory, Salk Institute, 92138 La Jolla, CA, USA |
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Abstract: | Summary A biotinylated P0 glycoprotein cDNA was hybridized in situ to aldehyde-fixed vibratome sections and to aldehyde-fixed thin sections of Lowicryl-embedded trigeminal ganglia of 15 day old rats. Alkaline phosphatase and peroxidase detectors were used for light microscopic (LM) studies and peroxidase or colloidal gold were employed for electron microscopic (EM) detection. In both LM and EM sections, probe was found in cytoplasmic areas of myelinforming Schwann cells that were enriched in granular endoplasmic reticulum, demonstrating that these regions contain P0 mRNA. Interestingly, P0 mRNA tended to cluster in regions close to the developing myelin sheath. Relatively simple methods are here described for EM detection of mRNA with reasonable tissue preservation and high resolution. These methods may be useful for developmental and disease-related studies of specific mRNAs in mammalian tissues. |
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