Abstract
In mouse, two types of monocilia found on cells comprising the node establish LR asymmetry. Motile node monocilia generate leftward flow (nodal flow) of the perinodal extraembryonic fluid. Immotile cilia containing polycystin-2 are mechanosensors, responding to nodal flow by increasing intracellular calcium in cells at the left of the node. We provide evidence for the nature of the midline barrier that limits the calcium signal to the left side of the node. E7.5 mouse embryos were cultured for 20 hrs in still culture with the gap junction blocker heptanol (3.5mM). Normal left-sided expression of Pitx2 was observed in 75%(n=36) control embryos and only 8%(n=12) embryos cultured in 3.5mM heptanol. Gap junctions at the node were visualized in fixed e7.75 embryos with antiCx43 antibodies. Abundant Cx43 containing gap junctions were at the margins and lateral to the node, and very few Cx43 containing gap junctions were within the node itself, or at the midline. Zebrafish embryos at the Kuppfer’s vesicle (KV) s...
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