Direct Visualization of Anesthesia-induced Subcellular Dysfunction in Non-Neuronal Tissues of C. elegans

Abstract

Volatile anaesthetics such as isoflurane, halothane, and sevoflurane are extensively utilized for their reversible induction of unconsciousness, exhibiting well-defined impacts on neural networks. However, their influence on non-neuronal and developing tissues is inadequately comprehended, generating concerns for vulnerable categories, including children and individuals receiving extended anesthesia. To rectify this deficiency, we examined the subcellular impacts of anaesthetics utilizing HEK 293T and HepG2 cell lines, alongside with Caenorhabditis elegans as a comprehensive organismal model. In vitro treatment to lidocaine and isoflurane resulted in mitochondrial depolarization, lysosomal aggregation, and buildup of reactive oxygen species (ROS). In vivo, 8% isoflurane induced mitochondrial fragmentation, loss of branching, and disruption of tubular lysosomes, indicating compromised energy metabolism and autophagic stress. We detected selective transcriptional inhibition of neuron- and immune-related promoters, although RNA Pol I remained active, indicating energy-conservation strategies. Systemic oxidative stress was validated by TIR-1::GFP aggregation and lipofuscin accumulation. Our research is one of the initial investigations that defines anaesthesia-induced subcellular dysfunction beyond the nervous system, providing novel insights for safer anaesthetic protocols and informing long-term health evaluations, especially in individuals who are developing or chronically exposed. Significance statementWhile volatile anaesthetics are widely used for inducing unconsciousness, their effects beyond neurons remain poorly understood. This study reveals that clinically used anaesthetics such as isoflurane and lidocaine cause profound subcellular dysfunction in non-neuronal systems, including mitochondrial depolarization, lysosomal disruption, transcriptional repression, and systemic oxidative stress. Using both human cell lines and Caenorhabditis elegans, we uncover conserved patterns of organellar damage and stress responses. These findings have critical implications for pediatric, intensive care, and long-term anaesthesia cases, urging the need to reassess anaesthetic safety profiles and develop biomarkers for systemic toxicity. Our work opens new avenues for designing cell-type-specific protective strategies under anaesthetic exposure.