Organization of the neuroepithelial actin cytoskeleton is regulated by heparan sulfation during neurulation

Abstract

Heparan sulfate and cytoskeletal actin microfilaments have both been shown to be important regulators of neural tube closure during embryonic development. To determine the functional relationship of these two molecules in formation of the spinal neural tube, we cultured ARC mouse embryos at embryonic day E8.5 in the presence of chlorate, a competitive inhibitor of glycosaminoglycan sulfation, and examined the effects on organization of actin microfilaments in the neuroepithelium. Compared against embryos cultured under control conditions, chlorate-treated embryos had shortened posterior neuropore, a loss of median hinge point formation and increased bending at the paired dorsolateral hinge points. Furthermore, apical organization of actin microfilaments in the neuroepithelial cells was absent, and this was associated with convex bending of the neuroepithelium. The results suggest that heparan sulfate is an important determinant of cytoskeletal actin organization during spinal neurulation, and that its biological action is dependent on sulfation of the heparan molecule.