Acta Neuropathologica

, Volume 84, Issue 6, pp 581–587

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

  • Helene Breitschopf
  • Gerda Suchanek
  • Robert M. Gould
  • David R. Colman
  • Hans Lassmann
Regular Papers

DOI: 10.1007/BF00227734

Cite this article as:
Breitschopf, H., Suchanek, G., Gould, R.M. et al. Acta Neuropathol (1992) 84: 581. doi:10.1007/BF00227734

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.

Key words

Non-radioactive in situ hybridization Proteolipid protein Myelin basic protein Myelin-associated glycoprotein 2′3′-Cyclic nucleotide 3′-phosphodiesterase 

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • Helene Breitschopf
    • 1
  • Gerda Suchanek
    • 1
  • Robert M. Gould
    • 2
  • David R. Colman
    • 3
    • 4
    • 5
  • Hans Lassmann
    • 1
    • 6
  1. 1.Research Unit for Experimental NeuropathologyAustrian Academy of SciencesViennaAustria
  2. 2.Department of PharmacologyNew York State Institute for Basic Research in Developmental DisabilitiesStaten IslandUSA
  3. 3.Department of AnatomyColumbia University College of Physicians and SurgeonsNew YorkUSA
  4. 4.Department of Cell BiologyColumbia University College of Physicians and SurgeonsNew YorkUSA
  5. 5.Department of PathologyColumbia University College of Physicians and SurgeonsNew YorkUSA
  6. 6.Neurological InstituteUniversity ViennaAustria