DISTRIBUTION OF DNA IN HUMAN SERTOLI-CELL NUCLEOLI
DISTRIBUTION OF DNA IN HUMAN SERTOLI-CELL NUCLEOLI
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DOI：
10.1177/41.10.7504007
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发表时间：
1993-10-01
影响因子：
3.2
通讯作者：
WACHTLER, F
中科院分区：
文献类型：
--
作者：
MOSGOLLER, W;SCHOFER, C;WACHTLER, F
为了更好地理解核仁的结构，不同的技术被用来定位DNA在致密纤维成分（DF）或纤维中心（FC）通过电子显微镜（EM）。由于哪些成分含有DNA仍然存在争议，我们使用各种方法研究了人类支持细胞中DNA的分布。以人总基因组DNA为探针，采用原位杂交法（ISH）和抗DNA抗体进行免疫金检测。这样就可以对单个分量的信号密度进行统计评估。feulgen样锇胺（OA）技术也被应用于DNA的选择性可视化。抗DNA抗体在线粒体、染色质和核仁DF中检测到DNA。使用人类总基因组DNA的ISH显示出类似的标记模式。我们得出结论：(a)利用胶体金的EM染色技术对DNA检测的敏感性似乎低于OA方法，(b) DF由不同分子组成的不同结构域组成，(c)去致密的DNA不一定局限于一个特定的核仁成分。
For better understanding of nucleolar architecture, different techniques have been used to localize DNA within the dense fibrillar component (DF) or within the fibrillar centers (FC) by electron microscopy (EM). Since it still remains controversial which components contain DNA, we investigated the distribution of DNA in human Sertoli cells using various approaches. In situ hybridization (ISH) with human total genomic DNA as probe and the use of anti-DNA antibody were followed by immunogold detection. This allowed statistical evaluation of the signal density over individual components. The Feulgen-like osmium-ammine (OA) technique for the selective visualization of DNA was also applied. The anti-DNA antibodies detected DNA in mitochondria, in chromatin, and in the DF of the nucleolus. ISH using human total genomic DNA showed similar labeling patterns. The OA technique revealed DNA filaments in the FC and focal agglomerates of decondensed DNA within the DE We conclude that (a) EM staining techniques that utilize colloidal gold appear to be less sensitive for DNA detection than the OA method, (b) the DF consists of different domains with different molecular composition, and (c) decondensed DNA is not necessarily confined to one particular nucleolar component.