Cilia play a key role in the determination of the left-right axis in vertebrates by generating and sensing flow of extraembryonic fluid at the left-right organizer (LRO). Perception of cilia-directed flow triggers a calcium signaling cascade which originates within the cilium itself and then is relayed into the surrounding mesendodermal tissue, thereby directing organ situs via the nodal pathway. Two types of cilia, motile and immotile, function simultaneously to coordinate and direct asymmetric intraciliary calcium signaling cues in the LRO. Here, we describe tools, reagents, and methodologies for the visualization and manipulation of both cilia types as well as intraciliary calcium signaling in the LRO of zebrafish.
Keywords: Cilium; GECI; Kupffer’s vesicle; Nodal cilia.