Differential expression of somatostatin receptors by quantitative PCR in the rat brain.
Differential expression of somatostatin receptors by quantitative PCR in the rat brain.
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发表时间：
1995-08
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通讯作者：
C. Viollet;A. Faivre‐Bauman;Ji Zhang;C. Llorens-Cortes;C. Loudes;C. Kordon;J. Epelbaum
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作者：
C. Viollet;A. Faivre‐Bauman;Ji Zhang;C. Llorens-Cortes;C. Loudes;C. Kordon;J. Epelbaum
生长抑素受体（sst）的五种亚型最近被克隆并报道在大鼠脑内表达。然而，常规的mRNA测量技术不允许准确地比较5 sst的表达水平。因此，我们建立了一个定量的逆转录聚合酶链反应方法为5 sst。通过在msst 1质粒中插入对应于用于扩增每个sst的特异性有义引物的序列来构建cRNA内标物。使用共同的反向引物，通过受体的独特引物对以相同的效率扩增野生型和标准RNA。该技术进行了验证，通过评估sst mRNA在3个脑结构中，不同的生长抑素受体结合水平先前报道。虽然区域之间的绝对表达水平相似，但sst 3是小脑中的主要亚型，sst 1在脊髓中占主导地位，sst 4和sst 2在下丘脑中同等表达。
Five subtypes of somatostatin receptors (sst) have recently been cloned and reported to be expressed in rat brain. However, conventional mRNA measurement techniques do not allow to accurately compare the levels of expression of the 5 sst. Thus, we established a quantitative reverse transcriptase polymerase chain reaction method for the 5 sst. A cRNA internal standard was constructed by inserting in msst1 plasmid sequences corresponding to specific sense primers for amplification of each sst. Using a common reverse primer, a unique primer pair by receptor amplifies both wild type and standard RNAs with the same efficiency. The technique was validated by evaluating sst mRNAs in 3 brain structures in which different somatostatin receptor binding levels were previously reported. While the absolute level of expression is similar between regions, sst3 is the major subtype in cerebellum, sst1 predominates in spinal cord and sst4 and sst2 are equally expressed in the hypothalamus.