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PMID: 1471468 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

In situ hybridization with digoxigenin-labeled probes: sensitive and reliable detection method applied to myelinating rat brain.

Acta neuropathologica ·Vol. 84 ·No. 6 ·1992-00-00 ·Pages 581-7

Breitschopf H, Suchanek G, Gould RM, Colman DR, Lassmann H

Abstract

A method for in situ hybridization of digoxigenin-labeled cDNA and cRNA probes to myelin protein mRNA is described. This technique has dual advantages of high structural resolution and high sensitivity and avoids problems associated with handling of radioactive materials. Furthermore, it can be readily combined in double labeling with immunocytochemical protein detection. We have used this technique to detect and locate mRNA for myelin basic protein (MBP), proteolipid protein (PLP), 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) and myelin-associated glycoprotein (MAG) in oligodendrocytes of 7-day-old and adult rat brains. PLP and MAG mRNA were restricted to the perinuclear cytoplasm, whereas MBP and CNPase mRNA was additionally present in peripheral oligodendrocyte processes.

MeSH Terms
Animals Brain Chemistry/physiology DNA Probes Digoxigenin/metabolism Hydrolysis Immunohistochemistry In Situ Hybridization Myelin Proteins/biosynthesis Myelin Sheath/physiology Protein Denaturation RNA Probes RNA, Messenger/metabolism Rats Rats, Sprague-Dawley Tissue Embedding Tissue Fixation
Chemicals
DNA Probes Myelin Proteins RNA Probes RNA, Messenger Digoxigenin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Breitschopf H
Research Unit for Experimental Neuropathology, Austrian Academy of Sciences, Vienna.
Suchanek G
Gould R M
Colman D R
Lassmann H
References (30)
30 references, click to expand
  1. Differential ultrastructural localization of myelin basic protein, myelin/oligodendroglial glycoprotein, and 2',3'-cyclic nucleotide 3'-phosphodiesterase in the CNS of adult rats.
    J Neurochem. 1989 Jan;52(1):296-304 PMID: 2462020
  2. Formation and detection of RNA-DNA hybrid molecules in cytological preparations.
    Proc Natl Acad Sci U S A. 1969 Jun;63(2):378-83 PMID: 4895535
  3. Cellular localization of induced human interferon-beta mRNA by non-radioactive in situ hybridization.
    Histochemistry. 1989;91(4):315-9 PMID: 2732096
  4. Small basic proteins of myelin from central and peripheral nervous systems are encoded by the same gene.
    Proc Natl Acad Sci U S A. 1986 Feb;83(4):1111-4 PMID: 2419899
  5. Synthesis and incorporation of myelin polypeptides into CNS myelin.
    J Cell Biol. 1982 Nov;95(2 Pt 1):598-608 PMID: 6183276
  6. Nucleotide sequences of two mRNAs for rat brain myelin proteolipid protein.
    Cell. 1985 Oct;42(3):931-9 PMID: 2414013
  7. Axons modulate myelin protein messenger RNA levels during central nervous system myelination in vivo.
    J Neurosci Res. 1990 Aug;26(4):409-18 PMID: 1700137
  8. Heavy metal intensification of DAB-based HRP reaction product.
    J Histochem Cytochem. 1981 Jun;29(6):775 PMID: 7252134
  9. Optimization of non-isotopic in situ hybridization on formalin-fixed, paraffin-embedded material using digoxigenin-labelled probes and transgenic tissues.
    J Pathol. 1992 May;167(1):9-17 PMID: 1625064
  10. Induction of stress protein HSP70 in nerve cells after status epilepticus in the rat.
    Neurosci Lett. 1989 May 22;100(1-3):259-64 PMID: 2761775
  11. Spatial segregation of mRNA encoding myelin-specific proteins.
    Proc Natl Acad Sci U S A. 1987 Nov;84(21):7773-7 PMID: 3478726
  12. Detection of viral sequences of low reiteration frequency by in situ hybridization.
    Proc Natl Acad Sci U S A. 1978 Dec;75(12):6125-9 PMID: 216014
  13. Acetylation of chromosome squashes of Drosophila melanogaster decreases the background in autoradiographs from hybridization with [125I]-labeled RNA.
    J Histochem Cytochem. 1978 Aug;26(8):677-9 PMID: 99471
  14. Localization of neuron-specific enolase (NSE) mRNA in human brain.
    Neurosci Lett. 1987 May 6;76(2):233-8 PMID: 3587757
  15. Combination of immunocytochemistry and in situ hybridization in the same tissue section of rat pituitary.
    J Histochem Cytochem. 1986 Jan;34(1):39-43 PMID: 3510246
  16. Expression of the 2',3'-cyclic nucleotide 3'-phosphohydrolase gene and immunoreactive protein in oligodendrocytes as revealed by in situ hybridization and immunofluorescence.
    Glia. 1988;1(3):184-90 PMID: 2852171
  17. Model system for optimising mRNA non-isotopic in situ hybridisation: riboprobe detection of lysozyme mRNA in archival gut biopsy specimens.
    J Clin Pathol. 1991 Oct;44(10):835-9 PMID: 1960217
  18. Molecular cloning of a 2',3'-cyclic nucleotide 3'-phosphodiesterase: mRNAs with different 5' ends encode the same set of proteins in nervous and lymphoid tissues.
    J Neurosci. 1987 Sep;7(9):2703-10 PMID: 3040924
  19. In situ hybridization analysis of myelin gene transcripts in developing mouse spinal cord.
    J Neurosci. 1989 Jan;9(1):248-57 PMID: 2464047
  20. Non-isotopic RNA probes. Comparison between different labels and detection systems.
    Histochemistry. 1989;93(2):191-6 PMID: 2613556
  21. Biosynthesis and insertion of Wolfgram protein into optic nerve membranes.
    Neurochem Res. 1985 Jul;10(7):897-907 PMID: 4047284
  22. The amino acid sequences of the myelin-associated glycoproteins: homology to the immunoglobulin gene superfamily.
    J Cell Biol. 1987 Apr;104(4):957-65 PMID: 2435742
  23. Biosynthesis of the myelin 2',3'-cyclic nucleotide 3'-phosphodiesterases.
    J Neurochem. 1990 Feb;54(2):656-61 PMID: 1688921
  24. In situ hybridisation.
    Ann Rheum Dis. 1990 Jun;49 Suppl 1:434-9 PMID: 2197995
  25. Relation between axons and oligodendroglial cells during initial myelination. I. The glial unit.
    J Neurocytol. 1990 Jun;19(3):313-28 PMID: 2391536
  26. Cellular and subcellular distribution of 2',3'-cyclic nucleotide 3'-phosphodiesterase and its mRNA in the rat central nervous system.
    J Neurochem. 1988 Sep;51(3):859-68 PMID: 2842456
  27. Simultaneous in situ detection of viral RNA and antigens.
    Proc Natl Acad Sci U S A. 1984 Sep;81(17):5445-8 PMID: 6206496
  28. Regional expression of myelin protein genes in the developing mouse brain: in situ hybridization studies.
    J Neurosci Res. 1988 Oct-Dec;21(2-4):238-48 PMID: 2464076
  29. Localization of mRNAs by in situ hybridization.
    Methods Cell Biol. 1991;35:37-71 PMID: 1779862
  30. Quantitative analysis of in situ hybridization methods for the detection of actin gene expression.
    Nucleic Acids Res. 1985 Mar 11;13(5):1777-99 PMID: 3889842
Article Info
Journal
Acta neuropathologica
Abbr.
Acta Neuropathol
ISSN
0001-6322
DOI
10.1007/BF00227734
Published
1992-00-00
Pages
581-7
Language
English
Region
Germany
NLM ID
0412041
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00227734 View journal details
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