A single protocol to detect transcripts of various types and expression levels in neural tissue and cultured cells: in situ ... View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1993-12

AUTHORS

Nicole Schaeren-Wiemers, Andrea Gerfin-Moser

ABSTRACT

We have developed a simple non-radioactive in situ hybridization procedure for tissue sections and cultured cells using digoxigenin-labelled cRNA probes. This protocol can be applied for the detection of various transcripts present at a wide range of expression levels in the central nervous system. Cerebellar hybridization signals for transcripts estimated to be expressed at high (MBP, myelin basic protein), moderate (GluR1, subunit of AMPA/kainate sensitive glutamate receptors) and low (inositol polyphosphate-5-phosphatase) levels of abundance are demonstrated as examples. The sensitivity and cellular resolution were significantly improved by avoiding any ethanol treatment commonly used in other procedures. The localization of a labelled cell with respect to its environment is shown to be more easily assessed by counterstaining of the tissue with the nuclear dye Hoechst 33258. The present protocol can be combined with immunocytochemistry as demonstrated for glial fibrillary acidic protein (GFAP). All steps of the procedure, including preparation and labelling of the cRNA probes, pretreatment of tissue, hybridization and visualization of the labelled transcripts, are described in detail. More... »

PAGES

431-440

Journal

TITLE

Histochemistry and Cell Biology

ISSUE

6

VOLUME

100

Author Affiliations

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/bf00267823

    DOI

    http://dx.doi.org/10.1007/bf00267823

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1052611363

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/7512949


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