Removal of cellular water prevents the reformation of the interphase nucleus

Abstract

The mature snRNP (small nuclear ribonucleoprotein) particles are localized quantitatively in the interphase nucleus. Like many nuclear antigens, they distribute throughout the cytoplasm after the nuclear envelope breaks down during mitosis and then return to the newly formed daughter nuclei in early G1. Their abundance and stability and the availability of monoclonal antibodies that recognize them, make the snRNP particles a useful model system for studying the reformation of the nucleus at the completion of mitosis. A wide variety of metabolic inhibitors and alterations in normal culture conditions were investigated for their ability to interfere with the return of the snRNP particles to daughter nuclei after mitosis. None of the well-characterized cytoskeletal inhibitors, biosynthetic inhibitors, calcium antagonists, nor ionophores were effective in interfering with this return. However, the removal of cellular water by exposure of cells to hypertonic medium during mitosis blocked the reformation of the nucleus and trapped the snRNP particles in the cytoplasm. In medium of twice the normal tonicity, the function of the mitotic spindle and the cleavage furrow are inhibited, however, the cells reattach to the substratum as if returning to interphase. The chromatin stays condensed and does not form a normal interphase nucleus and the snRNP particles stay dispersed throughout the cytoplasm. This condition is reversible and after return to normal medium the nucleus reforms and the snRNP particles collect in the new nuclei. After gentle extraction of metaphase cells, about 30% of the snRNP particles are soluble, however, the remainder are associated with an insoluble remnant. These data are consistent with the notion that the snRNP particles accumulate in the nucleus due to both preferential solubility and specific binding sites in the interphase nucleus.