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Amoebae of strain CL of Physarum polycephalum undergo apogamic development to form multinucleate plasmodia. During the amoebal-plasmodial transition, large uninucleate cells become irreversibly committed to plasmodium development. The transformation of an amoeba to a plasmodium involves a change in the tubulin isotypes expressed and a radical restructuring of cellular microtubules. During the transition the amoebal cytoplasmic microtubules, centrioles and cytoplasmic MTOC must disappear and the plasmodial-specific tubulin isotypes and intranuclear microtubule organising centre (MTOC) must be acquired. In developing cultures, amoebae lose the ability to flagellate before they become committed. Enriched suspensions of committed cells can be obtained by inducing asynchronous differentiating cultures to flagellate and passing the cells through a glass bead column. The resulting committed cells can be cultured, with some synchrony, to form plasmodia on bacterial lawns or in axenic liquid medium but cannot be cultured on axenic agar medium. During mating, cells lose the ability to flagellate early in plasmodium development. Committed cells from mating mixtures can be enriched in a similar way to committed cells of CL and have similar growth characteristics. Uninucleate committed cells of CL have the same DNA content as amoebae and plasmodia but have 6-10 times the amount of RNA. Apogamic committed cells express tubulin isotypes characteristic of amoebae, but after culture in axenic liquid medium, the cells express plasmodial-specific tubulin isotypes. Results suggest that plasmodial-specific tubulin isotypes are switched on in quadrinucleate cells. The amoebal cytoskeleton persists in binucleate and quadrinucleate cells but has disappeared in larger multinucleate cells. Mitosis in uninucleate committed cells is intranuclear (plasmodial-type). The amoebal MTOCs are eliminated during the first few mitotic cycles after commitment and do not become the plasmodial intranuclear MTOCs. Centriole loss apparently occurs before MTOC loss. |
