Chromatin structure in situ, as visualized in thin sections selectively stained for DNA

Abstract

The ultrastructural localization of [3H]uridine-labelled RNA synthesized in the course of the cell cycle of synchronized CHO cells is studied using high resolution autoradiography combined with a differential staining for nucleoproteins. It is shown that sites of RNA transcription can already be visualized on the periphery of chromosomes of apparently late metaphase-early anaphase cells with no visible association with the reforming nuclear membrane. In interphase cells they are associated with the border of intranucleolar chromatin or condensed nucleoplasmic chromatin, wherever this localization is made possible by the degree of chromatin dispersion. In cells labeled for 1 or 3 h, the rate of RNA synthesis is higher in S and G2 than in G1. When cells fixed immediately after a [3H]uridine pulse are compared with those post-incubated for 13 h in isotope-free medium, there is a clear difference in intensity of labelling between the nucleus and the cytoplasm. However, the localization pattern of radioactive RNA in the nucleus is similar for all incubation periods as well as for all phases of interphase. The groups of interchromatin granules are generally labeled weakly with radioactivity associated rather with the periphery of their clusters, or remain unlabelled. These results are discussed in the context of other recent findings concerning the distribution of RNA and RNP-structures in the nucleus.