Abstract:
Actin flow refers to the motion of the F-actin cytoskeleton and has been observed in many different cell types, especially in motile cells including neuronal growth cones. The direction of the actin flow is generally retrograde from the periphery toward the center of the cell. Actin flow can be harnessed for forward movement of the cell through substrate-cytoskeletal coupling; thus, a key function of actin flow is in cell locomotion. In this chapter, we illustrate three different methods of quantifying retrograde F-actin flow in growth cones derived from cultured Aplysia bag cell neurons. These methods include tracking the movement of surface marker beads as well as kymograph analysis of time-lapse sequences acquired by differential interference contrast (DIC) imaging or fluorescent speckle microscopy (FSM). Due to their large size, Aplysia neuronal growth cones are uniquely suited for these methods; however, they can also be applied to any other growth cones with clear F-actin-rich peripheral domains.
摘要:
肌动蛋白流指的是F-肌动蛋白细胞骨架的运动,并已在许多不同的细胞类型中观察到,尤其是在活动细胞中,包括神经元生长锥。肌动蛋白流的方向通常从细胞的外围向中心逆行。肌动蛋白流可通过底物-细胞骨架偶联用于细胞的向前运动;因此,肌动蛋白流的一个关键功能是细胞运动。在这一章中,我们说明了三种不同的方法来定量从培养的Aplysia袋细胞神经元衍生的生长锥中的逆行F-肌动蛋白流。这些方法包括跟踪表面标记珠的移动以及通过差分干涉对比(DIC)成像或荧光斑点显微镜(FSM)获得的延时序列的测速分析。由于尺寸大,Aplysia神经元生长锥是唯一适合这些方法;然而,它们也可以应用于具有清晰的富含F-肌动蛋白的外周结构域的任何其他生长锥。