Ning Zhang-Yong, Zhao De-Ming, Yang Jian-Min, Cui Ya-Li, Meng Li-Ping, Wu Chang-De, Liu Hong-Xiang
National Animal Transmissible Spongiform Encephalopathies Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing, PR China.
Anim Biotechnol. 2005;16(1):55-65. doi: 10.1081/abio-200053404.
Determination of tissue-specific expression of cellular prion protein (PrPc) is essential for understanding its poorly explained role in organisms. Herein we report on quantification of PrP mRNA in golden hamsters, a popular experimental model for studying mechanisms of transmissible spongiform encephalopathies (TSE), by real-time RT-PCR. Total RNA was isolated from four different regions of the brain and six peripheral organs of eight golden hamsters. PrP mRNA copy numbers were determined using absolute standard curve method with real-time quantitative PCR instrument. It was found that high mRNA levels were present in all four regions of the brain examined, including obex, neocortex, cerebellum, and thalamus. In peripheral organs examined, inguinal lymph node showed high level of the expression similar to that in overall brain; spleen, heart, liver, and lung showed moderate levels of the expression; and kidney showed the lowest expression. Our result is consistent with the potential involvement of different organs in prion diseases and offers essential data for further study of TSE mechanism in this animal model.
确定细胞朊蛋白（PrPc）的组织特异性表达对于理解其在生物体中难以解释的作用至关重要。在此，我们报告通过实时逆转录聚合酶链反应（RT-PCR）对金黄仓鼠（一种用于研究传染性海绵状脑病（TSE）机制的常用实验模型）中PrP mRNA进行定量分析。从八只金黄仓鼠的大脑四个不同区域和六个外周器官中分离总RNA。使用绝对标准曲线法和实时定量PCR仪器测定PrP mRNA拷贝数。结果发现，在所检测的大脑所有四个区域（包括延髓、新皮层、小脑和丘脑）中均存在高mRNA水平。在所检测的外周器官中，腹股沟淋巴结的表达水平较高，与全脑相似；脾脏、心脏、肝脏和肺表现出中等水平的表达；而肾脏的表达最低。我们的结果与不同器官可能参与朊病毒疾病一致，并为进一步研究该动物模型中的TSE机制提供了重要数据。
