Cytokinesis requires the interplay between the cytoskeleton and plasma membrane. Emerging evidence indicates that some cytokinetic components are essential for the postcytokinetic events such as epithelium organization and neural development. We have recently developed live cell imaging and conditional knockout techniques to visualize cytokinetic proteins in Caenorhabditis elegans Q neuroblasts and separate their postcytokinetic functions from cytokinetic ones. Here we describe how the fluorescent reporter strains and conditional knockout C. elegans are generated and how live cell imaging of Q neuroblast development are performed in our laboratory. Using these protocols, we uncovered a novel role of Anillin in stabilizing the actin network during neuronal migration and neurite outgrowth, and the postcytokinetic fate of midbody, which is released into the extracellular space and degraded by the adjacent macrophage using an apoptotic mimicry. These protocols could also be applicable to study other cellular processes in C. elegans or adapted to other model organisms, to provide better insights into their developmental basis.
Keywords: Anillin; Caenorhabditis elegans; Conditional knockout; Cytokinesis; Live cell imaging; Midbody; Neuroblast; Postcytokinetic; Somatic CRISPR-Cas9.
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