Kellerman K A, McNally J G
Department of Biology and the Institute for Biomedical Computing, Washington University, St. Louis, Missouri, 63130, USA.
Dev Biol. 1999 Apr 15;208(2):416-29. doi: 10.1006/dbio.1999.9208.
To examine the mechanisms of cell locomotion within a three-dimensional (3-D) cell mass, we have undertaken a systematic 3-D analysis of individual cell movements in the Dictyostelium mound, the first 3-D structure to form during development of the fruiting body. We used time-lapse deconvolution microscopy to examine two strains whose motion represents endpoints on the spectrum of motile behaviors that we have observed in mounds. In AX-2 mounds, cell motion is slow and trajectories are a combination of random and radial, compared to KAX-3, in which motion is fivefold faster and most trajectories are rotational. Although radial or rotational motion was correlated with the optical-density wave patterns present in each strain, we also found small but significant subpopulations of cells that moved differently from the majority, demonstrating that optical-density waves are at best insufficient to explain all motile behavior in mounds. In examining morphogenesis in these strains, we noted that AX-2 mounds tended to culminate directly to a fruiting body, whereas KAX-3 mounds first formed a migratory slug. By altering buffering conditions we could interchange these behaviors and then found that mound-cell motions also changed accordingly. This demonstrates a correlation between mound-cell motion and subsequent development, but it is not obligatory. Chimeric mounds composed of only 10% KAX-3 cells and 90% AX-2 cells exhibited rotational motion, suggesting that a diffusible molecule induces rotation, but many of these mounds still culminated directly, demonstrating that rotational motion does not always lead to slug migration. Our observations provide a detailed analysis of cell motion for two distinct modes of mound and slug formation in Dictyostelium.
为了研究三维(3-D)细胞团内细胞运动的机制,我们对盘基网柄菌菌丘中单个细胞的运动进行了系统的三维分析,菌丘是子实体发育过程中形成的第一个三维结构。我们使用延时去卷积显微镜检查了两个菌株,它们的运动代表了我们在菌丘中观察到的运动行为谱的两个端点。与KAX-3菌株相比,AX-2菌丘中的细胞运动缓慢,轨迹是随机和径向运动的组合,而在KAX-3菌株中,细胞运动速度快五倍,大多数轨迹是旋转运动。尽管径向或旋转运动与每个菌株中存在的光密度波模式相关,但我们也发现了一小部分但数量可观的细胞亚群,它们的运动方式与大多数细胞不同,这表明光密度波充其量不足以解释菌丘中的所有运动行为。在研究这些菌株的形态发生时,我们注意到AX-2菌丘倾向于直接发育成子实体,而KAX-3菌丘首先形成迁移性蛞蝓体。通过改变缓冲条件,我们可以互换这些行为,然后发现菌丘细胞的运动也相应改变。这表明菌丘细胞运动与后续发育之间存在相关性,但并非必然。仅由10%的KAX-3细胞和90%的AX-2细胞组成的嵌合菌丘表现出旋转运动,这表明一种可扩散分子诱导了旋转,但许多这样的菌丘仍然直接发育,这表明旋转运动并不总是导致蛞蝓体迁移。我们的观察结果对盘基网柄菌中菌丘和蛞蝓体形成的两种不同模式的细胞运动进行了详细分析。