Resolving forebrain developmental organisation by analysis of differential growth patterns

The forebrain is the most complex region of the vertebrate central nervous system, and its developmental organisation is controversial. We fate-mapped the embryonic chick anterior neural tube and built a 4D model of brain growth. We reveal modular patterns of anisotropic growth, ascribed to progenitor regions through multiplex hybridisation chain reaction. Morphogenesis is dominated by directional growth towards the eye, more isometric expansion of the prethalamus and dorsal telencephalon, and anterior movement of ventral cells into the hypothalamus. Comparative gene expression analysis and cell mixing experiments suggest the existence of a contiguous transverse boundary region, encompassing the zona limitans intrathalamica and retromammillary hypothalamus, that divides the anterior and posterior forebrain, and becomes distorted at the base of the zona limitans intrathalamica. Fate conversion experiments indicate that the hypothalamus is topologically tripartite, lying ventral to the telencephalon, prethalamus and zona limitans intrathalamica. Our findings challenge the widely accepted prosomere model of forebrain organisation, do not support a segmented anterior forebrain, and instead suggest a ‘tripartite hypothalamus’ model.