用核糖核酸酶保护法研究大鼠离体肾脏中促红细胞生成素mRNA水平的氧依赖性调节。
Oxygen-dependent modulation of erythropoietin mRNA levels in isolated rat kidneys studied by RNase protection.
作者信息
Ratcliffe P J, Jones R W, Phillips R E, Nicholls L G, Bell J I
机构信息
Nuffield Department of Clinical Medicine, John Radcliffe Hospital, Headington, Oxford, United Kingdom.
出版信息
J Exp Med. 1990 Aug 1;172(2):657-60. doi: 10.1084/jem.172.2.657.
Abstract
Using oligonucleotide primers complementary to conserved regions in the mouse erythropoietin (Epo) gene, a portion of the rat Epo gene was amplified by the polymerase chain reaction to produce a probe suitable for assay of rat Epo mRNA by RNAse protection. The assay, which has sufficient sensitivity to measure to Epo mRNA in unstimulated rat kidneys, was used to demonstrate high amplitude in vitro modulation of Epo mRNA levels in response to changes in perfusate flow rate and oxygen tension in isolated kidneys, thus providing clear evidence that all the necessary events linking changes in oxygen delivery to the modulation of Epo mRNA levels can occur intrarenally.
摘要
利用与小鼠促红细胞生成素（Epo）基因保守区域互补的寡核苷酸引物，通过聚合酶链反应扩增大鼠Epo基因的一部分，以产生适合通过RNA酶保护法检测大鼠Epo mRNA的探针。该检测方法具有足够的灵敏度，能够测量未受刺激的大鼠肾脏中的Epo mRNA，用于证明在离体肾脏中，Epo mRNA水平会因灌注液流速和氧张力的变化而在体外发生高幅度调节，从而提供了明确的证据，表明将氧输送变化与Epo mRNA水平调节联系起来的所有必要事件都可在肾脏内发生。
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