In situ hybridization with digoxigenin-labeled probes: sensitive and reliable detection method applied to myelinating rat brain.
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.
Breitschopf
H
H
Research Unit for Experimental Neuropathology, Austrian Academy of Sciences, Vienna.
Suchanek
G
G
Gould
R M
RM
Colman
D R
DR
Lassmann
H
H
D000818:Animals; D001923:Brain Chemistry / Q000502:physiology*; D015342:DNA Probes; D004076:Digoxigenin / Q000378:metabolism*; D006868:Hydrolysis; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D009185:Myelin Proteins / Q000096:biosynthesis; D009186:Myelin Sheath / Q000502:physiology*; D011489:Protein Denaturation; D015347:RNA Probes; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D016610:Tissue Embedding; D016707:Tissue Fixation
D015342:DNA Probes; D009185:Myelin Proteins; D015347:RNA Probes; D012333:RNA, Messenger; D004076:Digoxigenin