Generation of thalamic neurons from mouse embryonic stem cells

Abstract

The 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 generating thalamic neurons from mouse embryonic stem cells (mESCs) by modifying the default method that induces the most-anterior neural type in self-organizing culture. A low concentration of the caudalizing factor insulin and a MAPK/ERK kinase inhibitor enhanced the expression of the caudal forebrain markers Otx2 and Pax6. BMP7 promoted an increase in thalamic precursors such as Tcf7l2 + /Gbx2 + and Tcf7l2 + / Olig3 + cells. mESC thalamic precursors began to express the glutamate transporter vGlut2 and the axon-specific marker VGF, similar to mature projection neurons. The mESC thalamic neurons extended their axons to cortical layers in both 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.

TGFβ superfamily signaling regulates the state of human stem cell pluripotency and competency to create telencephalic organoids

“…One out of seven KSR tested batches (~14%) was optimal for producing forebrain organoids (data not shown), which was consistent with the previously reported mouse cerebral organoid formation (Nasu et al, 2012). Insulin has been shown to caudalize the early neuroectoderm and induce formation of the diencephalon, midbrain, and rostral hindbrain during organoid differentiation (Wataya et al, 2008;Muguruma et al, 2015;Shiraishi et al, 2017). However, insulin's roles in hPSC transcriptomic signatures have not yet been explored.…”

Neurons Generated by Mouse ESCs with Hippocampal or Cortical Identity Display Distinct Projection Patterns When Co-transplanted in the Adult Brain

“…So far, the regional identity of the neurons produced in vitro through the neuralization of pluripotent cells has mainly been established by their molecular characterization through variable degrees of analysis of their gene expression, ranging from the simple study of their neurotransmitter phenotype ( Eiraku et al., 2011 , Shi et al., 2012 , Shiraishi et al., 2017 , Yu et al., 2014 ) to a deeper investigation of their molecular nature by methods of global gene expression analysis ( Bertacchi et al., 2013 , Bertacchi et al., 2015a , Bertacchi et al., 2015b , Edri et al., 2015 , Espuny-Camacho et al., 2013 , Van de Leemput et al., 2014 , Yao et al., 2017 ). Even so, ascertaining the identity of a nerve cell produced in vitro by comparison of its global gene expression profile with that of neurons in vivo is very useful but not sufficient.…”

Brain Organoids as Model Systems for Genetic Neurodevelopmental Disorders

“…Indeed, self-patterning of organoids precursors can be restricted to forebrain fate by inhibiting TGF-β, BMP, and WNT pathways ( Figure 1A ; Eiraku et al, 2008 ; Kadoshima et al, 2013 ). By supplying additional patterning factors (such as WNT3A, SHH, insulin, and BMP7), forebrain fate can be further confined to produce organoids resembling any discrete part of the forebrain region, including cerebral cortex, optic cup, hippocampal, choroid plexus, subpallium, thalamus, and hypothalamus ( Wataya et al, 2008 ; Eiraku et al, 2011 ; Nakano et al, 2012 ; Germain et al, 2013 ; Kadoshima et al, 2013 ; Liu et al, 2013 ; Maroof et al, 2013 ; Nicholas et al, 2013 ; Mariani et al, 2015 ; Sakaguchi et al, 2015 ; Qian et al, 2016 ; Shiraishi et al, 2017 ; Kim et al, 2019b ; Xiang et al, 2020a ). Midbrain organoids are generated by combining TGF-β and BMP inhibition with WNT and SHH activation, and FGF8 treatment ( Jo et al, 2016 ; Kim et al, 2019a ), whereas cerebellum organoids are produced by timed and combinatory treatment with a series of patterning factors, including TGF-β and BMP inhibitors, insulin, FGF2, FGF19, and SDF1 ( Figure 1A ; Muguruma et al, 2015 ).…”