Tissue-specific expression pattern of bovine prion gene: quantification using real-time RT-PCR.
In recent studies PrP mRNA was determined mostly by in situ hybridisation or Northern Blot analysis--methods not suitable for absolute quantification of mRNA copy numbers. Herein we report on bovine prion mRNA quantification using calibrated highly sensitive externally standardized real-time RT-PCR with LightCycler instrument. Total RNA was isolated from nine different regions of the CNS and seven peripheral organs. PrP(c) mRNA copy numbers could be determined in all tissues under study. In approval with prior studies high mRNA level was found in Neocortex and Cerebellum. Lymphatic organs showed at least as high expression levels of prion mRNA as overall brain. Lowest expression was detected in kidney. Results of our study provide insight into the involvement of different organs in pathogenesis with respect to prion mRNA expression. LightCycler technology is currently considered the most precise method for nucleic acid quantification and showed to be powerful tool for further studies on prion diseases pathogenesis.
Tichopad
Ales
A
Institute of Physiology, FML Weihenstephan, Technical University of Munich, Weihenstephaner Berg 3, 85354 Freising, Germany.
Pfaffl
Michael W
MW
Didier
Andrea
A
D000818:Animals; D002417:Cattle; D002490:Central Nervous System / Q000378:metabolism; D008297:Male; D011328:Prions / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014018:Tissue Distribution
D011328:Prions; D012333:RNA, Messenger