A combined ultrastructural approach to the study of nuclear matrix thermal stabilization

Histochemistry. 1992 Sep;98(2):121-9. doi: 10.1007/BF00717003.

Abstract

Using mouse erythroleukaemia cells and different ultrastructural techniques, the morphology was investigated of the nuclear matrix obtained after incubation at 37 degrees C of isolated nuclei. If purified nuclei were heated for 45 min at 37 degrees C, the final matrix exhibited well-recognizable nucleolar remnants, an inner network and a peripheral lamina. Without such incubation only the peripheral lamina was seen surrounding homogeneous, finely granular material. Similar results were obtained with both araldite-embedded and freeze-fractured nuclear matrices, although in the latter case the loose granular material was not evident. Observations of araldite-embedded, heat-treated nuclei revealed clumping of heterochromatin in small, very electron-dense masses with large interchromatin spaces. These ultrastructural aspects were even more striking in freeze-fractured nuclei. Cytochemical matrix analysis by osmium-amine staining for nucleic acids and DNase-gold labelling for DNA localization demonstrated that also matrix residual nucleic acids, mostly RNA, are stabilized by heat exposure of isolated nuclei. The results demonstrate that the morphology of heat-stabilized nuclear matrix is not artefactually affected during the preparation for conventional electron microscopy and suggest a possible involvement of nucleic acids in the heat-induced stabilization of the nuclear matrix.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA, Neoplasm / metabolism
  • Deoxyribonucleases / metabolism
  • Freeze Fracturing
  • Friend murine leukemia virus*
  • Histocytochemistry
  • Leukemia, Erythroblastic, Acute / pathology*
  • Mice
  • Nuclear Matrix / ultrastructure*
  • Plastic Embedding
  • RNA, Neoplasm / metabolism
  • Staining and Labeling
  • Tumor Cells, Cultured

Substances

  • DNA, Neoplasm
  • RNA, Neoplasm
  • Deoxyribonucleases