Journal of Cell Science, Vol 16, 309-331, Copyright © 1974 by Company of Biologists

Submitted on November 9, 1973
Revised on June 10, 1974

Ultrastructure of Mitotic Arrest Induced by Isoprenaline in Rat Parotid Acinar Cells

¹ Biological Research Unit, Cancer Institute, 278 William Street, Melbourne 3000, Australia

The mitotic arrest of acinar cells in the rat parotid gland in response to isoprenaline has been investigated at the ultrastructural level. The arrested cells were characterized by the presence of both centriole pairs at the cell centre, around which chromosomes formed an approximately spherical array. Microtubules radiated out from both centriole pairs, indicating that they were part of a bipolar system. The microtubule population included both non-kinetochore and kinetochore microtubules. Metaphase arrest appeared to be due to failure of the poles to separate, or to remain separated in the case of cells already in metaphase at the time of drug administration. This could be explained by the drug interfering with the ability of interpolar microtubules from opposite poles to interact with one another.

The distribution of the chromosomes around the centrioles appears to depend on the presence of kinetochore microtubules, which are thought to act as tethers, either by themselves or in conjunction with non-kinetochore microtubules. It is suggested that tension is necessary between the kinetochores and poles to attain the arrest configuration. Chromosomes were observed in which both kinetochores had attached microtubules but whether or not they were linked to opposite poles remains to be investigated.

Two types of arrested cell were distinguished by their content of secretory granules. Cells already in mitosis at the time of isoprenaline administration were not depleted by drug action, and during the period of arrest numerous large secretory granules were intermingled with the chromosomes. Those cells which entered mitosis after the drug was given were initially blocked in the G₂ phase, during which time they were depleted of secretory granules. On entering mitosis, these cells possessed only small secretory granules, which were newly synthesized. In the metaphase-arrested state the small secretory granules were clustered around the centriolar complexes, demonstrating the presence of a poleward force operating on them. It is possible that the same force acts on the secretory granules in the first type of arrested cell, but because of the relatively large size of the granules, the clustering around the poles is not pronounced.

The nature of the forces acting on the chromosomes and on the secretory granules is discussed. The functioning of these forces during the arrest state has to be considered in any general model for mitosis.