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

## TL;DR

This study reveals how astrocytes change at the molecular level during wakefulness, sleep, and general anesthesia, uncovering shared and distinct patterns that may explain differences in consciousness.

## Contribution

The study identifies state-specific astrocytic proteomic and phosphoproteomic signatures, offering new insights into astrocytic roles in regulating consciousness.

## Key findings

- Sleep and anesthesia share similar proteomic changes but differ in molecular expression, particularly in metabolism and clearance processes.
- General anesthesia uniquely alters phosphorylation of NUCKS1 at Ser188, potentially suppressing nuclear transcription and cell cycle activity.
- Sleep is linked to upregulated mRNA processing, while anesthesia suggests enhanced synaptic signaling and suppressed developmental programs.

## 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.