Fakan S, Hughes M E
Centre of Electron Microscopy, University of Lausanne, Switzerland.
Chromosoma. 1989 Oct;98(4):242-9. doi: 10.1007/BF00327309.
The fine structure of the nuclear components was studied following mild lysis of mouse or Drosophila tissue culture cells and spreading of nuclear material. Particular attention was paid to nuclear ribonucleoprotein (RNP) constituents, which were analysed by high resolution autoradiography after [3H]uridine pulse labelling of cells. Comparison with the labelling kinetics of various in situ nuclear RNP constituents described previously suggests strong similarities between in situ constituents and structures observed within spread nuclear components. The present observations suggest that the nucleolar dense fibrillar component, shown previously in ultrathin sections of [3H]uridine-labelled intact cells as carrying rapidly labelled pre-rRNA, in fact consists of highly compacted transcribing ribosomal genes. The growing RNP fibrils appearing in transcription complexes of extranucleolar active genes and the in situ observed perichromatin fibrils also show the same labelling properties. This confirms that the two structures indeed represent the same nucleoplasmic constituents. As for the nuclear structures involved in post-transcriptional events, our observations demonstrate the occurrence of a rapidly labelled RNP fibro-granular network. Its granular elements correspond, in size and perichromatin location, to the perichromatin granules seen in the situ preparations and suggest similarities between the two constituents. The results are discussed in the light of other data providing information on the role of various nuclear structural constituents.
在对小鼠或果蝇组织培养细胞进行轻度裂解并铺展核物质后,研究了核成分的精细结构。特别关注了核糖核蛋白(RNP)成分,在对细胞进行[3H]尿苷脉冲标记后,通过高分辨率放射自显影对其进行了分析。与先前描述的各种原位核RNP成分的标记动力学进行比较表明,原位成分与在铺展的核成分中观察到的结构之间存在很强的相似性。目前的观察结果表明,核仁致密纤维成分,先前在[3H]尿苷标记的完整细胞的超薄切片中显示携带快速标记的前体rRNA,实际上由高度压缩的转录核糖体基因组成。出现在核仁外活性基因转录复合物中的正在生长的RNP纤维以及原位观察到的染色质周围纤维也显示出相同的标记特性。这证实了这两种结构确实代表相同的核质成分。至于参与转录后事件的核结构,我们的观察结果表明存在一种快速标记的RNP纤维颗粒网络。其颗粒成分在大小和染色质周围位置上与原位制备中看到的染色质周围颗粒相对应,表明这两种成分之间存在相似性。根据提供有关各种核结构成分作用信息的其他数据对结果进行了讨论。