Hranilović D, Lesch K P, Ugarković D, Cicin-Sain L, Jernej B
Department of Organic Chemistry and Biochemistry, Ruder Bosković Institute, Zagreb, Croatia.
J Neural Transm (Vienna). 1996;103(8-9):957-65. doi: 10.1007/BF01291786.
Total RNA isolated from rat platelets by guanidinium-acid-phenol extraction, and mRNA for the serotonin (5-hydroxytryptamine) transporter (5HTt) was identified. From a typical starting sample of 20 mL of rat blood (approximately 9 x 10(9) platelets), 14 to 17 micrograms of total platelet RNA was obtained. Northern blot analysis, using 32P-labeled 5HTt cDNA as a probe, identified approximately 3.3 kb long 5HTt mRNA. After rehybridization with cyclophilin cDNA, approximately 1 kb long mRNA for cyclophilin, which could serve as a reference for 5HTt mRNA quantification, was also identified. Densitometric analysis demonstrated clearly measurable signals for both mRNAs. The possibility of quantification of rat platelet 5HTt mRNA should enable parallel studies on 5HTt gene expression in platelets and brain of the same animal, and the evaluation of the platelet model at the molecular genetic level.
通过胍盐-酸性酚抽提法从大鼠血小板中分离出总RNA，并鉴定出5-羟色胺（5-羟色胺）转运体（5HTt）的mRNA。从20 mL大鼠血液（约9×10⁹个血小板）的典型起始样本中，获得了14至17微克的血小板总RNA。使用32P标记的5HTt cDNA作为探针进行Northern印迹分析，鉴定出约3.3 kb长的5HTt mRNA。在用亲环蛋白cDNA重新杂交后，还鉴定出了约1 kb长的亲环蛋白mRNA，它可作为5HTt mRNA定量的参照。密度分析清楚地显示了两种mRNA都有可测量的信号。对大鼠血小板5HTt mRNA进行定量的可能性，应能使我们对同一动物的血小板和脑中5HTt基因表达进行平行研究，并在分子遗传学水平上评估血小板模型。
