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Journal of Histochemistry and Cytochemistry, Vol. 51, 913-919, July 2003, Copyright © 2003, The Histochemical Society, Inc.

ARTICLE

Detection of Oligodendrocytes in Tissue Sections Using PCR Synthesis of Digoxigenin-labeled Probes

Walid Jalabia, Mirela Cerghetb, Robert P. Skoffb, and M. Said Ghandoura
a UMR 7004 CNRS/ULP, Faculté de Médecine, Université Louis Pasteur Strasbourg, France
b Department of Anatomy and Cell Biology, School of Medicine, Wayne State University, Detroit, Michigan

Correspondence to: M. Said Ghandour, UMR 7004 ULP/CNRS, Faculté de Médecine, 11 rue Humann, 67085 Strasbourg, France. E-mail: ghandour@neurochem.u-strasbg.fr

Oligodendrocytes, the myelin-forming cells in the central nervous system, were visualized with excellent resolution at the light microscopic level using in situ hybridization (ISH). Digoxigenin (Dig)-tagged probes were synthesized and efficiently labeled by PCR. Specific probes to myelin genes were made by RT from brain total RNAs, followed by PCR with designed specific primers in the presence of Dig-11-dUTP. Probes specific to proteolipid protein (PLP), PLP and its isoform DM20 (PLP/DM20), and myelin oligodendrocyte glycoprotein (MOG) were synthesized and labeled. ISH was then applied on vibratomed tissue sections from mouse brains. Despite a low expression of MOG-specific and PLP-specific mRNAs in adult and newborn mouse brains, an oligodendrocyte population was detected. The specificity of Dig-labeled probes was confirmed with the double labeling of carbonic anhydrase II (CA II) and glial fibrillary acidic protein (GFAP) immunocytochemistry and ISH. This versatile and easy method for synthesis and labeling of specific probes to oligodendrocytes can be also applied to detect many other mRNAs in the nervous system and in other tissues. (J Histochem Cytochem 51:913–919, 2003)

Key Words: PLP, MOG, nonradioactive PCR labeling, high-resolution in situ, hybridization, oligodendrocyte, carbonic anhydrase II, GFAP, myelin


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