CHEMOTAXIS AND CHEMOKINESIS IN DICTYOSTELIUM

描述（由申请人提供）：我们的下一个计划的目标 这一时期的具体目标是双重的。首先，我们将完成我们的高分辨率 基本运动行为、自然波浪中的行为以及 对1）cAMP的空间梯度，2）模拟时间的行为反应 cAMP的波和3）10-6 cAMP的快速加入。 包括细胞表面的三维重建和运动分析， 细胞核、伪足和尾足，以及动态3D重建， GFP标记的F-肌动蛋白、肌球蛋白II、肌球蛋白IA、肌球蛋白IB和肌球蛋白I的运动分析 SCAR，使用3D-DIAScon。此外，我们将扩展此描述， 包括两个附加行为（丝状体覆盖和流式传输）， 在聚集过程中遵循单细胞趋化性。然后用这个 行为表型的武器库，我们将通过突变分析进行研究 肌球蛋白重链（MHC）去磷酸化、肌球蛋白调节 轻链（RLC）磷酸化，特异性MHC和RLC激酶，肌球蛋白 Is、SCAR和其他多种调控和细胞骨架元件在 运动和趋化性。这些研究的结果将 整合到独特的趋化性动态模型中， 根据它们出现的波的相位， 以及它们引发的特定行为反应。

DESCRIPTION (provided by applicant): Our objectives for the next program period specific aims are, twofold. First, we will complete our high resolution 3D description of basic motile behavior, behavior in natural waves, and the behavioral responses to 1) a spatial gradient of cAMP, 2) simulated temporal waves of cAMP and 3) the rapid addition of 10-6 cAMP This description will include the 3D reconstructions and motion analysis of the cell surface, nucleus, pseudopods and uropods of cells, and dynamic 3D reconstruction and motion analysis of GFP-tagged F-actin, myosin II, myosin IA, myosin IB and SCAR, using 3D-DIAScon. In addition, we will expand this description to include two additional behaviors (filopodial override, and streaming) that follow single cell chemotaxis in the aggregation process. Then, using this arsenal of behavioral phenotypes, we will investigate through mutant analysis the roles of myosin heavy chain (MHC) dephosphorylation, myosin regulatory light chain (RLC) phosphorylation, specific MHC and RLC kinases, the myosin Is, SCAR, and a variety of additional regulatory and cytoskeletal elements in locomotion and chemotaxis. The results of these studies will then be incorporated into unique dynamic models of chemotaxis that identify independent pathways according to the phase of the wave from which they emerge and the specific behavioral response they elicit.