Clonal identity determines astrocyte cortical heterogeneity.

Astrocytes are the most numerous cell type in the brain, where they are known to play multiple important functions. While there is increasing evidence of their morphological, molecular, and functional heterogeneity, it is not clear whether their positional and morphological identities are specified during brain development. We address this problem with a novel strategy to analyze cell lineages through the combinatorial expression of fluorescent proteins. Following in utero electroporation, stochastic expression of these proteins produces inheritable marks that enable the long-term in vivo tracing of glial progenitor lineages. Analyses of clonal dispersion in the adult cortex revealed unanticipated and highly specific clonal distribution patterns. In addition to the existence of clonal arrangements in specific domains, we found that different classes of astrocytes emerge from different clones. This reinforces the view that lineage origin impinges on cell heterogeneity, unveiling a new level of astrocyte diversity likely associated with specific regional functions.