State-Dependent Remodeling of Astrocytic Proteome and Phosphorylation Signaling Networks Across Wake, Sleep, and General Anesthesia.

Astrocytes undergo extensive molecular reprogramming during transitions of consciousness. Proteomic and phosphoproteomic analyses of rat cortical astrocytes across wake, sleep, and sevoflurane-induced general anesthesia revealed state-specific molecular signatures. Sleep and anesthesia shared similar changes such as downregulated structural proteins and upregulated membrane transport complexes, but diverged in molecular expression. Anesthesia specifically suggested potential activation of cellular differentiation and structural plasticity pathways, while implying disruption of metabolism and molecular clearance compared to sleep. Phosphoproteomics identified downregulated phosphorylation of NUCKS1 at Ser188 during anesthesia, suggesting suppressed nuclear transcription or cell cycle activity as a potential molecular signature of the anesthetic state. Sleep was associated with upregulated mRNA processing, while anesthesia indicated potential enhancement of synaptic signaling and suppression of development-related programs.

Study at a glance

Abstract

Astrocytes critically regulate states of consciousness, yet their molecular profiles across wake, sleep, and general anesthesia remain unclear. This study conducted proteomic and phosphoproteomic analyses of rat cortical astrocytes across these states using sevoflurane. Data quality was validated using principal component analysis (PCA) and Pearson correlation coefficient (PCC). Proteomics showed state-specific signatures: sleep and anesthesia shared similar changes (downregulated structural proteins, upregulated membrane transport complexes) but diverged in molecular expression. Anesthesia specifically suggested potential activation of cellular differentiation/structural plasticity-related pathways but implied potential disruption of metabolism and molecular clearance processes compared to sleep. Phosphoproteomics revealed the unique phosphorylation changes during general anesthesia compared to wake and normal sleep: downregulated phosphorylation of nuclear casein kinase and cyclin-dependent kinase substrate 1 (NUCKS1) at Ser188, suggesting the potential suppression of nuclear transcription and/or cell cycle activity, which may act as a potential molecular signature associated with the anesthetic state. Clustering analysis showed that sleep was associated with upregulated mRNA processing, while anesthesia indicated potential enhancement of synaptic signaling and suggested possible suppression of development-related programs. In summary, astrocytes undergo extensive molecular reprogramming during transitions of consciousness; while they share common features in morphological remodeling, sleep and anesthesia differ fundamentally in astrocytic molecular outcomes, offering new insights into astrocytic roles in unconsciousness.