CHARACTERIZATION OF THE INTERCHROMATIN REGION AS THE NUCLEAR DOMAIN-CONTAINING SNRNPS IN PLANT-CELLS - A CYTOCHEMICAL AND IMMUNOELECTRON MICROSCOPY STUDY

The combination of electron microscopy (EM) cytochemical with immunocytochemical methods is used to characterize the interchromatin region (IR) of the plant cell nucleus. Cryoprocessing of the samples provides a better ultrastructural preservation and allows the observation of some differences in the fine structure of the IR which shows a denser aspect resulting from the lower extraction of components with low-temperature methods. A complex network of fibrillar structures and isolated or clustered 30 to 50-nm granules are observed in the IR. Anti-DNA antibodies combined with the NAMA-Ur method for DNA or the EDTA staining, preferential for RNPs, allow the detection of chromatin fibers in the IR. Bismuth staining reveals the presence of highly phosphorylated proteins in some interchromatin structures. The spliceosomal snRNPs are immunolocalized on cryosections and Lowicryl sections of plant cells using monoclonal and polyclonal antibodies. They provide a homogeneous immunofluorescence pattern with no speckles. This is in correlation with the labeling at EM, immunogold particles decorate the EDTA-positive fibrillar structures of the IR but no labeling is found over the 30 to 50-nm granules. The presence of the spliceosomal snRNPs, DNA and phosphorylated proteins in the IR indicate that this nuclear domain plays a major role in pre-messenger RNA splicing and, possibly in transcription, in the plant cell nucleus.