Perichromatin fibrils are in situ forms of nascent transcripts

The ultrastructural distribution of small nuclear ribonucleoproteins (U1-snRNP and Sm antigen) in the nucleus ofChironomus salivary gland cells was investigated by means of specific antibodies and immunocytochemistry using colloidal gold complexes as markers. Particular attention was paid to the structural relationships of snRNPs with transcriptionally active areas in polytene chromosomes and with Balbiani ring granules. Our results demonstrate strong binding of anti-snRNP antibodies to RNP fibrils or the fibrillar network occurring within nuclear regions active in transcription (Balbiani rings or minor puffs). This confirms data reported previously on mammalian cells showing early association of snRNPs with nuclear fibrillar constituents containing newly synthesized pre-mRNA. In addition, Balbiani ring granules in the process of formation at the transcriptionally active sites were sometimes labeled on their periphery or on the transitory part between the granule and its precursor RNP fibril, while free granules observed in the nucleoplasm were virtually devoid of label. These findings suggest that mRNA processing, including splicing, takes place prior to the formation of mature Balbiani ring granules.

Immunoelectron microscope localization of snRNPs in the polytene nucleus of salivary glands of Chironomus thummi.

The study of the cell nucleus using cryofixation and cryosubstitution

We studied the fine structural organization of the meristematic nucleus in roots of Lycopesicon esculentum (tomato) using ultracytochemical and immunocytochemical approaches. The nucleus has a non-reticulate (i.e. low DNA content) structure whose supramolecular organization differs in some respects from that in reticulate nuclei, principally in the organization of the chromocentres associated with the nuclear envelope, with which centromeric structures appear to be associated. The main difference at the nucleolar level is found in the fibrillar centres, which have a low amount of DNA labelling and in which inclusions of condensed chromatin are present only very rarely. The distribution of nucleolar DNA amongst the nucleolar compartments is similar to that in reticulate nucleoli as demonstrated using an anti-DNA monoclonal antibody. Tomato nuclei have nucleolus-associated bodies or karyosomes, like other plant species with a low DNA content and non-reticulate nuclear organization. The nuclear ribonucleoprotein structures in the inter- and perichromatin regions, namely inter- and perichromatin fibrils and granules, show similar ultrastructural and cytochemical characteristics in both types of nuclei.

Fine structural organization of a non-reticulate plant cell nucleus. An ultracytochemical and immunocytochemical study

The nucleolus is the main site for synthesis and processing of ribosomal RNA in eukaryotes. In mammals, plants, and yeast the nucleolus has been extensively characterized by electron microscopy, but in the majority of the unicellular eukaryotes no such studies have been performed. Here we used ultrastructural cytochemical and immunocytochemical techniques as well as three-dimensional reconstruction to analyze the nucleolus of Trypanosoma cruzi, which is an early divergent eukaryote of medical importance. In T. cruzi epimastigotes the nucleolus is a spherical intranuclear ribonucleoprotein organelle localized in a relatively central position within the nucleus. Dense fibrillar and granular components but not fibrillar centers were observed. In addition, nuclear bodies resembling Cajal bodies were observed associated to the nucleolus in the surrounding nucleoplasm. Our results provide additional morphological data to better understand the synthesis and processing of the ribosomal RNA in kinetoplastids.

https://www.cambridge.org/core/services/aop-cambridge-core/content/view/S1431927605050233

Electron microscopy analysis of the nucleolus of Trypanosoma cruzi.

The evolutionary variations of nuclear structure of animals, plants, fungi and protoctists were studied with electron microscopy by using techniques preferentially staining ribonucleoprotein (RNP) particles and chromatin. A remarkable similarity in the general morphological features of the RNP particles and chromatin arrangement is found in animals, plants and fungi. Important variations of these features were found in protoctists. These observations suggest that major evolutionary changes in the nuclear structure predate the acquisition of plastids by the ancestors of green plants. Once evolved, the nuclear structural pattern is conserved in plants and animals. Among protoctists studied, Kinetoplastida, Cryptomonadida and Volvocida have RNP particles and chromatin arrangement resembling those of plants and animals. These similarities may indicate a common ancestor. Important differences in the nuclear structure among Euglenida, Amebida, Cryptomonadida, Volvocida and Kinetoplastida support the view that Sarcomastigophora is a polyphyletic taxon. For the same reason Kinetoplastida and Euglenida must not be grouped in a monophyletic taxon. We propose that the variations of RNP particles may be related to the initial evolution of post-transcriptional processing.

Implications for evolution of nuclear structures of animals, plants, fungi and protoctists

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Implications for evolution of nuclear structures of animals, plants, fungi and protoctists