Related Experiment Video
Updated: Sep 27, 2026
Quantitation of Rabies Virus in Various Bovine Brain Structures
Published on: May 22, 2021
Tissue-specific expression pattern of bovine prion gene: quantification using real-time RT-PCR
Ales Tichopad1, Michael W Pfaffl, Andrea Didier
1Institute of Physiology, FML Weihenstephan, Technical University of Munich, Weihenstephaner Berg 3, 85354 Freising, Germany.
Abstract:
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.
Insights
This study quantifies bovine prion mRNA using real-time RT-PCR, revealing expression across various tissues. Lymphatic organs show high prion mRNA levels, comparable to the brain, offering insights into prion disease pathogenesis.
Area of Science:
- Veterinary Medicine
- Molecular Biology
- Neuroscience
Background:
- Prion mRNA quantification previously relied on imprecise methods like in situ hybridization and Northern Blot analysis.
- Accurate measurement of prion protein (PrP) mRNA levels is crucial for understanding prion-related diseases.
Purpose of the Study:
- To quantify bovine PrP mRNA copy numbers across various central nervous system (CNS) and peripheral organs.
- To establish a precise method for PrP mRNA quantification using real-time RT-PCR.
Main Methods:
- Isolation of total RNA from nine CNS regions and seven peripheral organs.
- Application of calibrated, externally standardized real-time RT-PCR (qRT-PCR) using the LightCycler instrument for absolute quantification of PrP mRNA.
Main Results:
- Bovine PrP mRNA was successfully quantified in all examined tissues.
- High PrP mRNA levels were observed in the Neocortex and Cerebellum, consistent with prior studies.
- Lymphatic organs exhibited prion mRNA expression levels as high as those in the brain, with the kidney showing the lowest expression.
Conclusions:
- Real-time RT-PCR with LightCycler technology provides a precise method for quantifying PrP mRNA in various bovine tissues.
- The widespread expression of PrP mRNA, particularly in lymphatic organs, suggests their involvement in prion pathogenesis.
- This quantification method is a powerful tool for future research into prion diseases.
