Villanueva José, Giménez-Molina Yolanda, Gutiérrez Luis M
Instituto de Neurociencias, Centro Mixto CSIC-Universidad Miguel Hernández, Alicante, Spain.
Methods Mol Biol. 2023;2565:297-309. doi: 10.1007/978-1-0716-2671-9_20.
Cultured bovine chromaffin cells have been characterized as a successful model to study changes in the cytoskeleton during the secretory process. In this sense, the distribution and dynamics of the F-actin cytoskeleton can be studied by confocal microscopy using appropriate molecular tools such as LifeAct, a peptide that stains the structures of F-actin. In this work, we describe some methodological protocols making possible to study, under controlled stimulus conditions, the local dynamic changes of F-actin in the cortical zone and also to detect the simultaneous displacements of chromaffin granules and organelles in active zones.
培养的牛嗜铬细胞已被视为研究分泌过程中细胞骨架变化的成功模型。从这个意义上说,可以通过共聚焦显微镜,使用合适的分子工具,如LifeAct(一种能对F-肌动蛋白结构进行染色的肽)来研究F-肌动蛋白细胞骨架的分布和动态变化。在这项工作中,我们描述了一些方法方案,使得在可控刺激条件下研究皮质区F-肌动蛋白的局部动态变化以及检测活性区嗜铬颗粒和细胞器的同步位移成为可能。