Direct Visualization of Anesthesia-Induced Subcellular Dysfunction in Caenorhabditis elegans

The volatile anesthetic isoflurane is extensively utilized for reversible induction of unconsciousness, exhibiting well-defined impacts on neural networks. However, its influence on the whole body in developing tissues remains inadequately understood. To rectify this deficiency, we examined the subcellular impacts of anesthetics utilizing Caenorhabditis elegans as a comprehensive in vivo organismal model. Our results demonstrated that isoflurane induced mitochondrial fragmentation, loss of branching, and disruption of tubular lysosomes. These observations suggested compromised energy metabolism and organellar stress. We further observed a selective reduction in GFP reporter fluorescence driven by neuron- and immune-related promoters, while core transcriptional machinery reporter signal from RPOA-2::GFP levels and nucleolar localization remained largely unchanged. This suggests that important transcriptional factors, such as RNA polymerase I, are maintained. Functionally, anesthesia-exposed worms exhibited increased mortality when challenged with the pathogens Pseudomonas aeruginosa, and while a similar trend was observed following Staphylococcus aureus challenge, it did not reach statistical significance, indicating compromised immune resilience. Systemic oxidative stress was validated by the H2DCFDA assay and lipofuscin accumulation. Our findings provide comprehensive insights into the organism-wide cellular stress induced by acute anesthetic exposure, laying the foundation for the development of safer anesthetic strategies.