Molecular diversity of diencephalic astrocytes reveals adult astrogenesis regulated by Smad4. (22nd September 2021) - Ohlig, Stefanie
Abstract: Astrocytes regulate brain‐wide functions and also show region‐specific differences, but little is known about how general and region‐specific functions are aligned at the single‐cell level. To explore this, we isolated adult mouse diencephalic astrocytes by ACSA‐2‐mediated magnetic‐activated cell sorting (MACS). Single‐cell RNA‐seq revealed 7 gene expression clusters of astrocytes, with 4 forming a supercluster. Within the supercluster, cells differed by gene expression related to ion homeostasis or metabolism, with the former sharing gene expression with other regions and the latter being restricted to specific regions. All clusters showed expression of proliferation‐related genes, and proliferation of diencephalic astrocytes was confirmed by immunostaining. Clonal analysis demonstrated low level of astrogenesis in the adult diencephalon, but not in cerebral cortex grey matter. This led to the identification of Smad4 as a key regulator of diencephalic astrocyte in vivo proliferation and in vitro neurosphere formation. Thus, astrocytes show diverse gene expression states related to distinct functions with some subsets being more widespread while others are more regionally restricted. However, all share low‐level proliferation revealing the novel concept of adult astrogenesis in the diencephalon. Synopsis: Here we show an unprecedented molecular diversity of diencephalic astrocytes with differences in similarity of gene expression in the forebrain. The expression of proliferation genes led to the novel concept of de novo astrogenesis with Smad4 as a key regulator of astrocyte proliferation. Subsets of diencephalic astrocytes show wide‐spread distribution in telencephalon. Subsets of astrocytes are highly region‐specific restricted in thalamic or hypothalamic regions. Diencephalic astrocytes proliferate at low levels in the adult brain generating new astrocytes. Small subsets of diencephalic astrocytes have NSC potential. Smad4 is a key regulator of astrocyte proliferation in the adult diencephalon. Abstract : Single cell RNA‐seq analysis identifies distinct subgroups of diencephalic astrocytes and finds that Smad4‐mediated astrogenesis occurs in the adult diencephalon.