从大鼠脑的小离散区域分离出在翻译和逆转录中高度活跃的游离和膜结合多核糖体及mRNA。

Isolation of free and membrane-bound polysomes and mRNA highly active in translation and reverse transcription from small discrete regions of rat brain.

作者信息

Murthy M R, Lévesque G, Pandian S, Viallard J L, Ogier R, Cavagna A M, Dastugue B

机构信息

Department of Biochemistry, Faculty of Medicine, Laval University, Québec, Canada G1K 7P4.

出版信息

Neurochem Int. 1986;8(3):381-7. doi: 10.1016/0197-0186(86)90011-2.

DOI:10.1016/0197-0186(86)90011-2

PMID:20493067

Abstract

A procedure is described for the preparation of total polysomes, membrane-bound and free polysomes and polysomal mRNA from as little as 5 mg or less of brain tissue. These preparations were highly active when tested for translation and reverse transcription in vitro. Using this method, polysomes and mRNA from rat cerebral cortex, cerebellum, hippocampus and hypothalamus were compared. The results showed that membrane-bound polysomes were more active than free polysomes in protein synthesis. The activities of polysomes and mRNA for protein and cDNA synthesis were dependent on the specific brain structures from which they were obtained. Polysomes from cerebellum and hypothalamus incorporated amino acids more actively than those from cerebral cortex or hippocampus, when tested in a reticulocyte lysate system. Cerebellar mRNA also showed the highest activity for cDNA syntehsis as compared to mRNAs from the other three tissues.

摘要

本文描述了一种从低至5毫克或更少的脑组织中制备总多核糖体、膜结合多核糖体和游离多核糖体以及多核糖体mRNA的方法。这些制备物在体外进行翻译和逆转录测试时具有很高的活性。利用该方法，对大鼠大脑皮层、小脑、海马体和下丘脑的多核糖体和mRNA进行了比较。结果表明，膜结合多核糖体在蛋白质合成中比游离多核糖体更具活性。多核糖体和mRNA用于蛋白质和cDNA合成的活性取决于它们所取自的特定脑结构。在网织红细胞裂解物系统中进行测试时，来自小脑和下丘脑的多核糖体比来自大脑皮层或海马体的多核糖体更积极地掺入氨基酸。与其他三个组织的mRNA相比，小脑mRNA在cDNA合成方面也表现出最高的活性。