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高分辨率放射自显影术作为一种用于定位哺乳动物细胞核中核酸合成与分布的工具。

High-resolution autoradiography as a tool for the localization of nucleic acid synthesis and distribution in the mammalian cell nucleus.

作者信息

Fakan S

出版信息

J Microsc. 1976 Mar;106(2):159-71. doi: 10.1111/j.1365-2818.1976.tb02398.x.

DOI:10.1111/j.1365-2818.1976.tb02398.x
PMID:58064
Abstract

Several examples of the application of high resolution autoradiography to the study of nucleic acid distribution in ultrathin sections of fixed and plastic embedded or frozen material are presented. Newly-synthesized DNA, labelled by very short pulses of 3H-thymidine is found to be localized throughout the nucleus. In the blastomeres of early mouse embryos developing after fertilization by 3H-thymidine-labelled spermatozoa, the labelled paternal DNA is distributed non-homogeneously in the nucleus. Finally some results obtained by using a cytochemical enzymatic technique for visualization of DNA directly in ultrathin sections of fixed and embedded cells are demonstrated. Concerning the distribution of RNA, perichromatin localization of rapidly transcribed RNA is described in different cell systems. This label can be attributed to perichromatin fibrils. In cells labelled for longer periods of time, sometimes followed by prolonged postincubations, a residual label is always found over the nucleus. Clusters of interchromatin granules are usually only weakly labelled or unlabelled. These structures probably contain a limited amount of rather slowly labelled RNA. The present results are discussed in the context of some biochemical evidence and of the data described by other investigators.

摘要

本文展示了高分辨率放射自显影技术在固定、塑料包埋或冷冻材料超薄切片中核酸分布研究的几个应用实例。用³H - 胸苷极短脉冲标记的新合成DNA被发现定位于整个细胞核。在用³H - 胸苷标记的精子受精后发育的早期小鼠胚胎的卵裂球中,标记的父本DNA在细胞核中分布不均匀。最后展示了一些通过细胞化学酶技术直接在固定和包埋细胞的超薄切片中可视化DNA所获得的结果。关于RNA的分布,在不同细胞系统中描述了快速转录RNA的染色质周边定位。这种标记可归因于染色质周边纤维。在标记较长时间的细胞中,有时随后进行长时间的孵育后,在细胞核上总是能发现残留标记。染色质间颗粒簇通常仅被弱标记或未被标记。这些结构可能含有有限量的标记相当缓慢的RNA。本文结果结合一些生化证据和其他研究者描述的数据进行了讨论。

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High-resolution autoradiography as a tool for the localization of nucleic acid synthesis and distribution in the mammalian cell nucleus.高分辨率放射自显影术作为一种用于定位哺乳动物细胞核中核酸合成与分布的工具。
J Microsc. 1976 Mar;106(2):159-71. doi: 10.1111/j.1365-2818.1976.tb02398.x.
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Fine structural cytochemical and immunocytochemical analysis of nucleic acids and ribonucleoprotein distribution in nuclei of pig oocytes and early preimplantation embryos.猪卵母细胞和植入前早期胚胎细胞核中核酸和核糖核蛋白分布的精细结构细胞化学和免疫细胞化学分析
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Robert Feulgen Prize Lecture 1995. New approaches to in situ detection of nucleic acids.1995年罗伯特·福尔根奖讲座。核酸原位检测的新方法。
Histochem Cell Biol. 1995 Aug;104(2):81-95. doi: 10.1007/BF01451570.
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New data concerning the functional organization of the mammalian cell nucleolus: detection of RNA and rRNA by in situ molecular immunocytochemistry.关于哺乳动物细胞核仁功能组织的新数据:通过原位分子免疫细胞化学检测RNA和rRNA。
Nucleic Acids Res. 1992 Dec 11;20(23):6195-200. doi: 10.1093/nar/20.23.6195.