Habenular Neurogenesis in Zebrafish Is Regulated by a Hedgehog, Pax6 Proneural Gene Cascade

The habenulae are highly conserved nuclei in the dorsal diencephalon that connect
the forebrain to the midbrain and hindbrain. These nuclei have been implicated in
a broad variety of behaviours in humans, primates, rodents and zebrafish. Despite
this, the molecular mechanisms that control the genesis and differentiation of
neural progenitors in the habenulae remain relatively unknown. We have previously
shown that, in zebrafish, the timing of habenular neurogenesis is left-right
asymmetric and that in the absence of Nodal signalling this asymmetry is lost.
Here, we show that habenular neurogenesis requires the homeobox transcription
factor Pax6a and the redundant action of two proneural bHLH factors, Neurog1 and
Neurod4. We present evidence that Hedgehog signalling is required for the
expression of pax6a, which is in turn necessary for the expression of neurog1 and
neurod4. Finally, we demonstrate by pharmacological inhibition that Hedgehog
signalling is required continuously during habenular neurogenesis and by cell
transplantation experiments that pathway activation is required cell
autonomously. Our data sheds light on the mechanism underlying habenular
development that may provide insights into how Nodal signalling imposes asymmetry
on the timing of habenular neurogenesis.