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

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Summary

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.

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Authors and Affiliations

  1. Research Unit for Experimental Neuropathology, Austrian Academy of Sciences, Schwarzspanierstraße 17, A-1090, Vienna, Austria

    Helene Breitschopf, Gerda Suchanek & Hans Lassmann

  2. Department of Pharmacology, New York State Institute for Basic Research in Developmental Disabilities, 10314, Staten Island, NY, USA

    Robert M. Gould

  3. Department of Anatomy, Columbia University College of Physicians and Surgeons, 10032, New York, NY, USA

    David R. Colman

  4. Department of Cell Biology, Columbia University College of Physicians and Surgeons, 10032, New York, NY, USA

    David R. Colman

  5. Department of Pathology, Columbia University College of Physicians and Surgeons, 10032, New York, NY, USA

    David R. Colman

  6. Neurological Institute, University Vienna, Austria

    Hans Lassmann

Authors
  1. Helene Breitschopf
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  2. Gerda Suchanek
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  3. Robert M. Gould
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  4. David R. Colman
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  5. Hans Lassmann
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Supported by the Science Research Fund, Austria, P 7740M

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Breitschopf, H., Suchanek, G., Gould, R.M. et al. In situ hybridization with digoxigenin-labeled probes: sensitive and reliable detection method applied to myelinating rat brain. Acta Neuropathol 84, 581–587 (1992). https://doi.org/10.1007/BF00227734

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  • DOI: https://doi.org/10.1007/BF00227734

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