Effect of Cerophyl growth medium on exocytosis in Tetrahymena thermophila.

Culturing the ciliate Tetrahymena thermophila in Cerophyl has provided an opportunity for studying the assembly and/or synthesis of the intramembrane particle array, the rosette, which marks the site of exocytosis in these cells. Cultures grown in this medium cease cell division after only 12h and enter 'stationary phase' earlier (12h of growth) relative to growth in standard medium (proteose peptone). In addition, the cell changes from the normally observed pear-shaped body to a thinner more ellipsoid form. Despite the initial similarities to starving cells, several differences are observed in the Cerophyl-grown cells. One is that cell size remains constant for at least 72h in contrast to starved cells. Secondly, in spite of this block in cell division, results from freeze-fracture replicas of the cell membrane of these cells show that they continue to assemble rosettes, the number of which increases approximately six times, from 0.34 rosette/microgram2 to 2.1 rosettes/microgram2. Addition of the protein synthesis inhibitor, cycloheximide (6h exposure), during growth in Cerophyl shows that 70% of rosettes can be assembled, despite the blockage of translation, by using pre-existing component(s) from a pool. The remaining 30% must involve de novo synthesis of one or more components; this percentage can be increased with longer exposure to the drug. Thirdly, an apparent increase in the number of mucocysts is observed by thin-section electron microscopy. At first (12-24h) only docked mucocysts seem to accumulate in the cell. However, by 36h a considerable increase seems to have taken place, particularly in the number of mucocysts located in the cytoplasm. In the cycloheximide-treated cells this increase in mucocysts begins to be blocked after 6h of exposure to the drug. These observations are in agreement with the results obtained from the freeze-fracture data on the concomitant increase in number of rosettes. This system therefore offers the first possibility of exploring the biosynthesis of these components.