Early role for a Na⁺,K⁺-ATPase (<i>ATP1A3</i>) in brain development.

Abstract

Osmotic equilibrium and membrane potential in animal cells depend on concentration gradients of sodium (Na⁺) and potassium (K⁺) ions across the plasma membrane, a function catalyzed by the Na⁺,K⁺-ATPase α-subunit. Here, we describe <i>ATP1A3</i> variants encoding dysfunctional α3-subunits in children affected by polymicrogyria, a developmental malformation of the cerebral cortex characterized by abnormal folding and laminar organization. To gain cell-biological insights into the spatiotemporal dynamics of prenatal <i>ATP1A3</i> expression, we built an <i>ATP1A3</i> transcriptional atlas of fetal cortical development using mRNA in situ hybridization and transcriptomic profiling of ∼125,000 individual cells with single-cell RNA sequencing (Drop-seq) from 11 areas of the midgestational human neocortex. We found that fetal expression of <i>ATP1A3</i> is most abundant to a subset of excitatory neurons carrying transcriptional signatures of the developing subplate, yet also maintains expression in nonneuronal cell populations. Moving forward a year in human development, we profiled ∼52,000 nuclei from four areas of an infant neocortex and show that <i>ATP1A3</i> expression persists throughout early postnatal development, most predominantly in inhibitory neurons, including parvalbumin interneurons in the frontal cortex. Finally, we discovered the heteromeric Na⁺,K⁺-ATPase pump complex may form nonredundant cell-type-specific α-β isoform combinations, including α3-β1 in excitatory neurons and α3-β2 in inhibitory neurons. Together, the developmental malformation phenotype of affected individuals and single-cell <i>ATP1A3</i> expression patterns point to a key role for α3 in human cortex development, as well as a cell-type basis for pre- and postnatal <i>ATP1A3</i>-associated diseases.