Dorsoventral patterning of the avian mesencephalon/metencephalon: Role of the notochord and floor plate in suppressing Engrailed‐2

Abstract

Transcription factors that are spatially and temporally restricted within the embryo may be used for dorsoventral and rostrocaudal positional information during development. The Engrailed-2 (En-2) gene is expressed across the mesencephalon/metencephalon (mes/met) boundary in the cerebellar primordium with strong dorsolateral expression and limited expression in the floor plate. In a previous experiment we demonstrated that, after removal of Hensen's node, embryos lacked a notochord in the head and the pattern of En-2 expression was normal rostrocaudally, but it was expanded into the ventral midline of the neural tube. This suggested that the notochord suppresses En-2 in the ventral neural tube during normal development. To test further the ability of the notochord (and floor plate) to suppress En-2, we transplanted ventral midline tissues from HH 5-9 quail embryos beneath the rostral neural plate of HH 4-6 chick embryos. After 24 hours in culture, 90% of the embryos with quail notochord or floor plate near the mes/met of the host lacked En-2 expression adjacent to the graft, and suppression was distance dependent. Enzymatically isolated notochords also suppressed En-2 (71%), but the results from isolated floor plates were inconclusive. Other grafts served as controls and included tissues from the trunk ventral midline, mes/met level dorsolateral neural plate, and trunk dorsolateral neural plate/somite. Collectively, the results suggest that during normal development the notochord and possibly the floor plate are important regulators of normal En-2 expression.