Development of a Three-dimensional Gel Culture Model of Caenorhabditis elegans Muscular Atrophy Induced by Acoustic Levitation Simulating Microgravity

Aims: This study aimed to establish an acoustic levitation-based microgravity simulation model and investigate its effects on the locomotor function and muscle protein expression of Caenorhabditis elegans. Methodology: L4-stage N2 nematodes were assessed on gel ball and on culture plate after 3-, 6-, 9-, and 12-hours post-inoculation. The acoustic levitation system was developed using the standing wave principle, with a three-dimensional gel ball culture system serving as the suspension carrier. Locomotor behavior including head swing frequency, bending frequency and swimming ability were observed after suspended for 9 hours, as well as muscle protein expression of RW1596 nematodes. Results: Nematodes were suspended for 9 hours, during which significant reductions were observed in head swing frequency (41.67 to 21.87 times/min, a 47.52% decrease), bending frequency (19.33 to 9.4 times/min, a 51.38% decrease), and swimming frequency (38.88 to 32.19 times/min, a 17.2% decrease) (all P < 0.01). Additionally, MYO-3 protein fluorescence intensity declined by 17.87% (P < 0.0001). Conclusion: These findings demonstrate that acoustic levitation can induce a muscle atrophy phenotype analogous to that seen in true microgravity within a short time frame. This approach avoids mechanical contact interference and provides a novel ground-based simulation platform for aerospace medical research.