Mitosis in Schizosaccharomyces pombe has been followed in living cells by phase-contrast microscopy and studied in fixed and suitably stained preparations by light microscopy. Successful preservation of nuclear fine structure in this yeast, not previously achieved, has allowed us to confirm and extend the observations made with light microscopy.
Without first arranging themselves on a metaphase plate, mitotic chromosomes become grouped in 2 clusters radiating, finger-like, from 2 points of attachment at opposite poles of an elongating nucleus. At these 2 sites electron microscopy reveals the presence of disk-shaped electron-dense organelles which we have called kinetochore equivalents (KCE). At mitosis the KCEs are connected across the nucleus by a narrow bundle of parallel microtubules which we refer to as the spindle.
Integration of our observations has led us to propose that at mitosis the separation of the KCEs and their attached chromosomes is initiated by a differential expansion of the nuclear envelope restricted to the region between recently divided KCEs and that expansion of the nuclear envelope later becomes general, resulting in a marked elongation of the nucleus. Displacement of the nuclear contents to the ends of the elongated nucleus gives it the shape of a dumbbell.
The elongation of the microtubule bundle keeps in step with the elongation of the nucleus but does not appear to be the cause of it. It may have the function of keeping the separated KCEs rigidly apart.
During mitosis the nucleolus persists and stretches out within the unbroken envelope of the nucleus as it elongates. Towards the end of division equal amounts of nucleolar material are found in the rounded ends of the dumbbell-shaped nucleus.
The break up of the dumbbell shape into daughter nuclei seems to involve the breaking of its tenuous middle part and a pivoting of its 2 ends in opposite directions.
In the course of our work on mitosis we have become aware of features in the cytoplasm of growing S. pombe cells which are described here for the first time. The cells invariably contain several prominent vacuoles containing an extremely electron-dense material which stains metachromatically with toluidine blue and may be polyphosphate. The mitochondria are of special interest for 2 reasons. First, because they have unique mesosome-like membrane invaginations and secondly, because a mitochondrion is regularly associated with the single KCE by the side of the interphase nucleus, as well as with each one of the 2 KCEs that occupy opposite ends of the intranuclear spindle during mitosis.