Generation of thalamic neurons from mouse embryonic stem cells [TECHNIQUES AND RESOURCES ARTICLE]

Atsushi Shiraishi, Keiko Muguruma, and Yoshiki Sasai

Thalamus is a diencephalic structure that plays crucial roles in relaying and modulating sensory and motor information to the neocortex. The thalamus develops in the dorsal part of the neural tube at the level of the caudal forebrain. However, the molecular mechanisms that are essential for thalamic differentiation are still unknown. Here we have succeeded in the generation of thalamic neurons from mouse ES cells (mESCs) by modifying the default method that induces the most-anterior neural type in self-organizing culture. A low concentration of caudalizing factor insulin and a MAPK/ERK kinase inhibitor enhanced the expression of caudal forebrain markers Otx2 and Pax6. BMP7 promoted an increase in the thalamic precursors such as Tcf7l2⁺/Gbx2⁺ and Tcf7l2⁺/Olig3⁺. mESC-thalamic precursors became to express the glutamate transporter vGlut2 and axon-specific marker VGF, similar to mature projection neurons. The mESC-thalamic neurons extended their axons to cortical layers both in organotypic culture and subcortical transplantation. Thus we have identified the minimum elements sufficient for in vitro generation of thalamic neurons. These findings expand our knowledge of thalamic development.

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