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来自哺乳动物大脑皮层的神经元细胞核和胶质细胞核。核小体重复长度、DNA含量和H1含量。

Neuronal nuclei and glial nuclei from mammalian cerebral cortex. Nucleosome repeat lengths, DNA contents and H1 contents.

作者信息

Pearson E C, Bates D L, Prospero T D, Thomas J O

出版信息

Eur J Biochem. 1984 Oct 15;144(2):353-60. doi: 10.1111/j.1432-1033.1984.tb08471.x.

Abstract

We have characterised the histone and DNA contents of neuronal and glial nuclei from ox cerebral cortex which have, respectively, repeat lengths of 162 base pairs and 201 base pairs. Although the neuronal population cannot be obtained completely free of glial nuclear contamination, the degree of contamination is easily determined by counting, and has been allowed for in all the methods used here. By diphenylamine assay and flow cytofluorometry we find that the DNA contents of both nuclear types are essentially equal, and equivalent to the diploid value, contrary to some reports. By quantification of the core histones in known numbers of nuclei with respect to an added external standard, we have shown that the ratio of core histone octamers in the two nuclear types, neuronal and glial, is the inverse of the ratio of repeat lengths. Thus the same proportion of DNA is associated with core histone octamers in the two nuclear types, most simply all of the DNA. By complete radiolabelling of the lysine side chains of the histones with methyl [1-3H]acetimidate we have determined the stoichiometry of H1 relative to the core histones. Neuronal nuclei have a low H1 content of 0.45 molecule H1/nucleosome on average; glial nuclei have the 'normal' 1 H1 molecule/nucleosome. In neuronal nuclei about half of the nucleosomes therefore probably lack H1. Whether there is any relation between the low H1 content and the short DNA repeat length of neuronal nuclei, on the one hand, and their high transcriptional capacity (at least when assayed in vitro), on the other, remains to be established.

摘要

我们已对牛大脑皮质中神经元和神经胶质细胞核的组蛋白和DNA含量进行了表征,其重复长度分别为162个碱基对和201个碱基对。尽管无法获得完全不含神经胶质细胞核污染的神经元群体,但通过计数很容易确定污染程度,并且本文所使用的所有方法均已考虑到这一点。通过二苯胺测定法和流式细胞荧光测定法,我们发现与一些报道相反,两种核类型的DNA含量基本相等,且等同于二倍体值。通过相对于添加的外部标准对已知数量细胞核中的核心组蛋白进行定量,我们表明神经元和神经胶质这两种核类型中核心组蛋白八聚体的比例与重复长度的比例呈反比。因此,两种核类型中与核心组蛋白八聚体结合的DNA比例相同,最简单的情况是所有DNA都如此。通过用甲基[1-³H]乙酰亚胺对组蛋白的赖氨酸侧链进行完全放射性标记,我们确定了H1相对于核心组蛋白的化学计量。神经元核平均每个核小体的H1含量较低,为0.45个H1分子;神经胶质核具有“正常的”每个核小体1个H1分子。因此,在神经元核中,大约一半的核小体可能缺乏H1。一方面,神经元核中低H1含量和短DNA重复长度与另一方面它们的高转录能力(至少在体外测定时)之间是否存在任何关系,仍有待确定。

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