CYTOCHEMICAL AND IMMUNOCYTOCHEMICAL STUDY OF NUCLEAR-STRUCTURES OF LACANDONIA-SCHISMATICA

The interphase nucleus of the cells of several tissues of Lacandonia schismatica was studied using electron microscopy cytochemical and immunocytochemical methods. The EDTA staining procedure, preferential for RNP, contrasted the Lacandonia granules and perichromatin fibrils. These granules were found to be relatively resistant to RNAse hydrolysis, but they were easily digested if RNAse treatment was carried out after a short hydrolysis with pronase. Bismuth oxynitrate stained granular structures about 17 nm in diameter and the periphery of a few Lacandonia granules. The anti-snURNPs bound to RNP-containing fibrils in the perichromatin and interchromatin space and also to the periphery of some Lacandonia granules. Immunolabeling of DNA demonstrated numerous filaments of extended chromatin in the perichromatin and interchromatin spaces which were closely related to, Lacandonia granules. These observations suggested that Lacandonia granules are equivalent to Balbiani ring granules of nuclei with polytene chromosomes and to perichromatin granules of other plant and animal nuclei. The small number of Lacandonia granules labeled in their periphery by anti-snURNP mAb were interpreted as being immature granules in the process of formation. The external or annular part of the ring-shaped structures is heavily labeled by anti-URNP mAbs but scarcely stained by the EDTA procedure. These features indicate that this region contains abundant proteins associated with snURNAs but probably little snURNAs. The synaptonemal-like complexes previously found in the interphase nuclei of Lacandonia are formed by two parallel masses of compact chromatin, which react with anti-DNA, and a central clear space crossed by fiber. Most of these fibers do not react with tests for DNA or RNA and are probably composed of proteins. The EDTA procedure contrasts some fibrils at the internal aspect of the masses of compact chromatin, suggesting the presence of remnants of the lateral element of a synaptonemal complex.