本研究旨在检测绵羊组织中朊病毒受体37kDa/67kDa LRP/LR mRNA水平并探讨其与朊病毒组织嗜性的关系。选用背景相似的6只内蒙绵羊,提取组织RNA,反转录RT-PCR构建cDNA模板;利用本研究前期构建的标准质粒及标准曲线,对组织中该受体mRNA水平进行Real-time(实时)荧光定量PCR检测。结果表明,大脑皮质中的受体表达水平最高(P〈0.05),其次为心脏和脑干,中等表达的器官依次为海马、小脑、脾脏、丘脑、肠系膜淋巴结、肝脏和肾脏,表达量最低为肺脏(P〈0.05)。结果提示,朊病毒受体——37kDa/67kDa LRP/LR在绵羊各组织中的表达量高低与朊病毒的复制程度有一定的相关性,受体LRP/LR表达量较高的区域朊病毒复制水平相对较高。结果提示朊病毒入侵机体后,组织器官PrPSc聚集程度可能与受体37kDa/67kDa LRP/LR表达水平相关。
The aim of the present study was to detect the mRNA transcription level of prion receptor--37kDa/67kDa LRP/LR in ovine tissues. The real-time RT-PCR was employed for the quantitation of mRNA transcription level of 11 tissues from six sheep with the same background. The synthesis of cDNA for each sample was firstly performanced by RT-PCR after the isolation of RNA from various tissues in sheep. The results showed that the highest LRP/LR mRNA transcription level was found in neocortex (P〈0.05) , which exhibiting as much as ten-fold higher than in lung, followed by heart and obex(P〈0.05) ; intermedia transcription was occured in hippocampus, cerebellum, spleen, thalamus, mesenteric lymph node; the lower transcription was oceured in liver and kidney, and the lung expressed extremely lowest but detectable level of LRP/ LR mRNA (P〈0.05). These results indicated that altered expression profiles of laminin receptor in various ovine normal tissues are basically corresponding to the degree of PrP^Sc accumulation, and tissues expressing high levels of LRP/LR may have higher potential susceptibility to PrP^Sc. Our results also suggest that in ovine tissues, the degree of PrPsc propagation was correlated with the expression level of prion receptor,37kDa/67kDa LRP/LR.