Finney R E, Mitchell L H, Soll D R, Murray B A, Loomis W F
Dev Biol. 1983 Aug;98(2):502-9. doi: 10.1016/0012-1606(83)90379-2.
When developing cultures of Dictyostelium discoideum are disaggregated and resuspended in nutrient medium, they lose the capacity to rapidly reaggregate after 90 min, in a rapid and synchronous step referred to as the "erasure event." They then proceed to lose remaining developmentally acquired functions in a program of dedifferentiation culuminating with the loss of EDTA-resistant cohesion roughly 5 hr later. Immediately following the erasure event, cells can be stimulated to reenter the developmental program even though they still possess a number of developmentally acquired functions. These cells therefore appear to undergo dedifferentiation and redifferentiation simultaneously (D. R. Soll and L. H. Mitchell, 1982, Dev. Biol. 91, 183-190). In this report, we have employed an antiserum made against a developmentally acquired membrane glycoprotein, gp80, to examine whether gp80 is lost during dedifferentiation and whether it is either reutilized or resynthesized during redifferentiation. Results are presented which demonstrate that (1) when 9-hr developing cells are disaggregated and resuspended in nutrient medium, gp80 continues to accumulate for several hours after the erasure event, then is lost at roughly the same time as EDTA-resistant cohesion; (2) when cells are stimulated to reenter the developmental program immediately after the erasure event, both gp80 and EDTA-resistant cohesion are still lost according to the program of dedifferentiation, but are then reacquired soon afterwards according to the program of redifferentiation; (3) during redifferentiation, cells do not reutilize gp80 which had been synthesized during initial development; rather they synthesize gp80 de novo; and (4) developing cells of a dedifferentiation-defective variant, HI4, when disaggregated and resuspended in nutrient medium, retain gp80, EDTA-resistant cohesion, and the capacity to rapidly reinitiate aggregation for at least 12 hr. This last result indicates that the loss of gp80 is regulated by the dedifferentiation process and is not an independent response to disaggregation or the reintroduction of nutrients. Together, these results reinforce the conclusion that dedifferentiation and redifferentiation can function independently and simultaneously in the same cells.
当将发育中的盘基网柄菌培养物分散并重新悬浮于营养培养基中时,它们在90分钟后会在一个被称为“擦除事件”的快速同步步骤中丧失快速重新聚集的能力。然后,它们会在一个去分化程序中逐渐丧失其余发育获得的功能,大约5小时后最终丧失对EDTA抗性的黏附力。在擦除事件之后,细胞即使仍具有许多发育获得的功能,也能被刺激重新进入发育程序。因此,这些细胞似乎同时经历去分化和再分化(D. R. 索尔和L. H. 米切尔,1982年,《发育生物学》91卷,第183 - 190页)。在本报告中,我们使用了一种针对发育获得的膜糖蛋白gp80制备的抗血清,来研究gp80在去分化过程中是否丢失,以及它在再分化过程中是否被重新利用或重新合成。结果表明:(1)当9小时龄的发育细胞被分散并重新悬浮于营养培养基中时,gp80在擦除事件后仍会积累数小时,然后与对EDTA抗性的黏附力大致同时丧失;(2)当细胞在擦除事件后立即被刺激重新进入发育程序时,gp80和对EDTA抗性的黏附力仍会按照去分化程序丧失,但随后会按照再分化程序很快重新获得;(3)在再分化过程中,细胞不会重新利用初始发育期间合成的gp80;相反,它们会重新合成gp80;(4)去分化缺陷变体HI4的发育细胞在被分散并重新悬浮于营养培养基中时,会保留gp80、对EDTA抗性的黏附力以及快速重新启动聚集的能力至少12小时。最后这个结果表明,gp80的丢失受去分化过程调控,而不是对分散或重新引入营养物质的独立反应。总之,这些结果强化了去分化和再分化可以在同一细胞中独立且同时发挥作用的结论。