Salcedo R M, Festoff B W, Citron B A
Department of Veterans Affairs Medical Center, Kansas City, MO, USA.
J Mol Neurosci. 1998 Apr;10(2):113-9. doi: 10.1007/BF02737122.
Thrombin acts on cells through the surface protease-activated receptor 1 (PAR-1), a G-protein-coupled member of the seven-transmembrane domain superfamily. On neural cells, thrombin has deleterious effects, killing neurons through apoptosis. Consequently, knowledge of PAR-1 expression in the nervous system may help to elucidate the role of thrombin in neurodegenerative disease. We developed a mimic construct to facilitate the highly sensitive technique of quantitative reverse transcriptase to PCR (qRT-PCR) to measure the differential expression of low copy number PAR-1 mRNA in neurodegenerative model systems. In this article, we report our results comparing homozygous wobbler (wr/wr) mice and normal littermates. By optimizing the transcription and quantitative PCR procedures to facilitate rapid copy number determination in small RNA samples, we documented a fivefold greater level of PAR-1 mRNA in the cervical spinal cord of wr/wr.
凝血酶通过表面蛋白酶激活受体1（PAR-1）作用于细胞，PAR-1是七跨膜结构域超家族的G蛋白偶联成员。在神经细胞上，凝血酶具有有害作用，通过凋亡杀死神经元。因此，了解PAR-1在神经系统中的表达情况可能有助于阐明凝血酶在神经退行性疾病中的作用。我们开发了一种模拟构建体，以促进定量逆转录聚合酶链反应（qRT-PCR）这一高灵敏度技术，来测量神经退行性模型系统中低拷贝数PAR-1 mRNA的差异表达。在本文中，我们报告了比较纯合摇摆（wr/wr）小鼠和正常同窝小鼠的结果。通过优化转录和定量PCR程序，以便在小RNA样本中快速确定拷贝数，我们记录到wr/wr小鼠颈脊髓中PAR-1 mRNA的水平高出五倍。
