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Light and electron microscopic localization of GABAA-receptors on cultured cerebellar granule cells and astrocytes using immunohistochemical techniques

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Abstract

GABAA-receptors were localized in explant cultures of rat cerebellum and in dissociated primary cultures of rat cerebellar granule cells and rat cerebellar astrocytes using the monoclonal antibody bd-17 directed against the β-subunit of the GABAA/benzodiazepine/chloride channel complex. At the light microscope level specific staining of GABAA-receptors was localized in various types of neurones in explant cultures of rat cerebellum using the indirect peroxidase-antiperoxidase (PAP) technique, whereas no specific staining was found in astrocytes. At the electron microscope level labeling of GABAA-receptors was observed in the plasma membrane of both the cell bodies and processes in dissociated primary cultures of cerebellar granule cells using an indirect preembedding immunogold staining technique which in contrast to the classical PAP technique allows quantitative estimations to be performed. Quantification of the labeling intensity revealed a higher concentration of GABAA-receptors per μm plasma membrane in the cell bodies than in the processes. In discrete areas an extremely high density of the GABAA-receptors was observed. No specific labeling of GABAA-receptors was observed in dissociated primary cultures of cerebellar astrocytes.

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Authors and Affiliations

  1. PharmaBiotec Research Center, Dept. of Biochemistry A, Panum Institute, Univ. of Copenhagen, DK-2200, Copenhagen N, Denmark

    Gert H. Hansen & Bo Belhage

  2. Department of Physiology, University of Basel, CH-4051, Basel, Switzerland

    Elisabeth Hösli & Leo Hösli

  3. PharmaBiotec Research Center, The Neurobiology Unit, Dept. of Biology B, Royal Danish School of Pharmacy, 2 Universitetsparken, DK-2100, Copenhagen Ø, Denmark

    Arne Schousboe

Authors
  1. Gert H. Hansen
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  2. Elisabeth Hösli
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  3. Bo Belhage
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  4. Arne Schousboe
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  5. Leo Hösli
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Special issue dedicated to Dr. Eugene Roberts.

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Hansen, G.H., Hösli, E., Belhage, B. et al. Light and electron microscopic localization of GABAA-receptors on cultured cerebellar granule cells and astrocytes using immunohistochemical techniques. Neurochem Res 16, 341–346 (1991). https://doi.org/10.1007/BF00966097

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