Distinct requirements for Ascl1 in subpopulations of midbrain GABAergic neurons.

Midbrain GABAergic neurons regulate multiple aspects of behavior and play important roles in psychiatric and neurological disease. These neurons constitute several anatomical and functional subpopulations, but their molecular heterogeneity and developmental regulatory mechanisms are poorly understood. Here we have studied the involvement of the proneural gene Ascl1 in the development of the midbrain GABAergic neurons. Analysis of Ascl1 mutant mice demonstrated highly region-specific requirements for Ascl1 for development of different GABAergic neuron subpopulations. Ascl1 is dispensable for the development of the ventral-most midbrain GABAergic neurons associated with dopaminergic nuclei substantia nigra pars reticulata (SNpr) and ventral tegmental area (VTA) GABAergic neurons. In the ventrolateral midbrain, loss of Ascl1 results in markedly delayed neurogenesis in the midbrain domains m3-m5. Within this region, Ascl1 has a unique role in m4, where it also regulates glutamatergic neurogenesis. Our results suggest that the m3-m5 midbrain neuroepithelium gives rise to the GABAergic neuron groups located in the midbrain reticular formation and ventrolateral periaqueductal gray. In contrast to m3-m5, Ascl1 is absolutely required in the dorsal midbrain domains m1-m2, for generation of the GABAergic neurons populating the superior and inferior colliculi as well as dorsal periaqueductal gray. These studies demonstrate different molecular regulatory mechanisms for the distinct midbrain GABAergic neuron subpopulations. Also, our results have implications on understanding the origins of the various midbrain GABAergic neuron groups in the embryonic neuroepithelium.

Analysis of the midbrain-hindbrain boundary cell fate using a boundary cell-specific Cre-mouse strain.

We describe here a transgenic mouse line MHB-Cre, which expresses Cre recombinase in a group of cells at the midbrain-hindbrain boundary. Using this mouse line, we studied the contribution of the boundary cells to distinct brain areas during development. Initially, the MHB-Cre expression coincides with that of Cdh22 and p21 around the Otx2 expression border in a narrow population of cells with reduced proliferative activity. Consistent with their location on both sides of the Otx2 expression border, the Cre expressing boundary cells contribute both to midbrain as well as hindbrain. However, the majority of recombinant cells remain close to the mid- and hindbrain border, suggesting very limited cell mixing within these brain compartments during development. Interestingly, dorsocaudally oriented fibers of the midbrain dopaminergic neurons follow the path marked by the boundary cells.

Gata2 is a tissue-specific post-mitotic selector gene for midbrain GABAergic neurons.

Midbrain GABAergic neurons control several aspects of behavior, but regulation of their development and diversity is poorly understood. Here, we further refine the midbrain regions active in GABAergic neurogenesis and show their correlation with the expression of the transcription factor Gata2. Using tissue-specific inactivation and ectopic expression, we show that Gata2 regulates GABAergic neuron development in the mouse midbrain, but not in rhombomere 1, where it is needed in the serotonergic lineage. Without Gata2, all the precursors in the embryonic midbrain fail to activate GABAergic neuron-specific gene expression and instead switch to a glutamatergic phenotype. Surprisingly, this fate switch is also observed throughout the neonatal midbrain, except for the GABAergic neurons located in the ventral dopaminergic nuclei, suggesting a distinct developmental pathway for these neurons. These studies identify Gata2 as an essential post-mitotic selector gene of the GABAergic neurotransmitter identity and demonstrate developmental heterogeneity of GABAergic neurons in the midbrain.