During vertebrate brain development, the onset of neuronal differentiation is under strict temporal control. In the mammalian thalamus and other brain regions, neurogenesis is regulated also in a spatially progressive manner referred to as a neurogenetic gradient, the underlying mechanism of which is unknown. Here we describe the existence of a neurogenetic gradient in the zebrafish thalamus and show that the progression of neurogenesis is controlled by dynamic expression of the bHLH repressor her6. Members of the Hes/Her family are known to regulate proneural genes, such as Neurogenin and Ascl. Here we find that Her6 determines not only the onset of neurogenesis but also the identity of thalamic neurons, marked by proneural and neurotransmitter gene expression: loss of Her6 leads to premature Neurogenin1-mediated genesis of glutamatergic (excitatory) neurons, whereas maintenance of Her6 leads to Ascl1-mediated production of GABAergic (inhibitory) neurons. Thus, the presence or absence of a single upstream regulator of proneural gene expression, Her6, leads to the establishment of discrete neuronal domains in the thalamus.diencephalon ͉ Hes1 ͉ mash1 ͉ ngn1 ͉ zona limitans intrathalamica N eurogenesis in the developing vertebrate CNS is regulated with a high degree of temporal and spatial precision, with stereotypic patterns of neuronal differentiation and extensive neuronal migration (1, 2). Dynamic patterns of mitotically active neuronal precursors, known as 'neurogenetic gradients' (3) have been described in several brain regions, including the neocortex (4, 5), the dorsal midbrain colliculi (6), and the dopaminergic region of the ventral midbrain (7). In the mammalian diencephalon, in particular the thalamus (formerly known as dorsal thalamus), two main neurogenetic gradients have been described: from posterior to anterior and from lateral to medial (8,9). In rodents, all thalamic neurons are generated in about 6 days, and the orthogonal gradients of glutamatergic neurogenesis sweep across the boundaries of future nuclei. The underlying molecular mechanisms responsible for generating the neurogenetic gradients of the thalamus are unknown.The major constituent of the thalamus is a population of excitatory neurons generated in the caudal thalamus (cTh), whereas a minor population of inhibitory neurons is generated in the rostral thalamus (rTh). The latter is thought to give rise to the reticular nucleus and the ventral lateral geniculate nucleus, including the intergeniculate leaflet (10). During development, this rostro-caudal partitioning is seen in the expression domains of proneural basic helix-loop-helix (bHLH) genes: the achaete-scute-like complex genes (Ascl formerly known as Mash in mouse and Zash in zebrafish) mark the GABAergic rTh and the prethalamus (PTh) and the Neurogenin genes (Neurog, formerly known as Ngn) mark the glutamatergic cTh (2,11,12). Several lines of evidence show that these genes function as determinants of transmitter phenotype: in the mouse telencephalon, Neurog1/2 are required to ...