High-dimensional analysis of single neurosphere molecular states reveals an ethanol-mediated shift in distribution along maturation trajectories

ABSTRACT: The fetal cerebral cortical neuroepithelium is a clonal mosaic, wherein populations of neural stem and daughter progenitor cells cycle asynchronously between multiple molecular states to generate cortical plate neurons. We hypothesized that these molecular states within neuroepithelial clonal colonies may be perturbed by fetal teratogens like alcohol, and that analyses of these perturbations may yield clues about mechanisms underlying teratogenesis. Fetal mouse dorsal telencephalon was microdissected at gestational day (GD) 12.5, dispersed single cells cultured for 5 days to yield clonal neurosphere cell aggregates, in either defined control culture media, or with a dose-range of ethanol. We assessed 68 gene transcripts by multiplexed RT-PCR in 264 individual neurospheres, across all treatment conditions. Cluster analysis resulted in the classification of control neurospheres into seven separate categories along a developmental trajectory from neuroepithelial, to transit-amplifying, to neuronal identity. Moreover, smaller control neurospheres were more immature than larger neurospheres. Ethanol exposure resulted in dose-related dissociation between neurosphere size and maturation state, with smaller neurospheres biased towards later maturation states. This pro-maturation effect was particularly pronounced at the highest doses of ethanol. These data suggest that ethanol facilitates neuroepithelial maturation with concurrent loss of stem cells, which may contribute to diminished brain growth.

ORGANISM(S): Mus musculus

PROVIDER: GSE167604 | GEO | 2026/02/02

REPOSITORIES: GEO