Behavioral Analysis of Locomotor Dysfunction in Drosophila melanogaster as a Readout for Neurotoxicity

This protocol presents a high-throughput, cost-effective method for assessing neurotoxicity in the fruit fly (Drosophila melanogaster) by quantifying locomotor dysfunction. Traditional mammalian models, while effective, are resource-intensive and ethically constrained, highlighting the need for alternative in vivo models. The present approach integrates the classic climbing assay with real-time monitoring platform to capture motor impairments in individual flies. This combined system enhances sensitivity, automation, and reproducibility, providing a scalable strategy for neurotoxicological screening. Critical procedural steps include standardized rearing conditions, careful anesthesia and handling to minimize fly mortality or behavioral alterations, and proper calibration of the system. Microcapillary-based feeding enables the optional integration of ingestion quantification, complementing the locomotor analysis and allowing the evaluation of compound intake when required. The method supports real-time and long-term behavioral monitoring, reduces observer bias, and enables daily progress evaluation without interrupting the experiment. Light preference testing can be incorporated using direct measurements. The platform provides significant advantages in behavioral resolution, throughput, and data consistency. Although it lacks real-time fluorescence capabilities found in some alternative systems, it remains highly effective for large-scale behavioral screening. This protocol is particularly valuable for laboratories aiming to assess the neurotoxic effects of pharmaceutical compounds, environmental agents, or genetic modifications using a sensitive, reproducible, and ethically favorable in vivo model.