Introduction: Ischemic stroke is a leading cause of motor and cognitive impairments, highlighting the need for effective model organisms to understand its underlying mechanisms. Drosophila melanogaster, with its well-characterized nervous system, rapid generational turnover, and high genetic similarity to humans, serves as a valuable model for studying ischemia-induced neurological deficits. Approximately 75% of human disease-associated genes and the entire hypoxia-induced cascade are conserved between flies and humans, making Drosophila particularly useful for investigating hypoxia’s effects on brain function. This study aims to assess motor and sensorycognitive impairments in Drosophila following hypoxic exposure, using behavioral assays such as chemotaxis in response to acetic acid, phototaxis, and locomotion . These assays allow for a multidimensional evaluation of sensory processing, decision-making, and motor coordination, providing insights into neurological deficits induced by stroke. Methods: Fly Preparation Wild-type Drosophila melanogaster (0-5 days old) were divided into control and experimental groups. Hypoxia was induced at 2.5, 4, and 6 hours using a device that reduces oxygen levels while maintaining environmental conditions. After recovery, behavior was assessed through various assays. Chemotaxis Assay A 20 cm vial divided into four areas was used to test chemotaxis. A cotton ball saturated with 5% acetic acid was placed at the top. Flies\\\\\\\\\\\\\\\' positions were recorded, focusing on their movement away from the acetic acid toward the farthest area. Locomotion Assay General motor activity was assessed in an arena where flies could walk freely without flying. Movements were tracked to evaluate total distance traveled, speed, and rest periods. Hypoxia-treated flies were compared with controls to assess motor deficits. Phototaxis Assay Two setups were used: 1. Dark Box Setup: Flies were placed in a dark box connected to a 20 cm vial divided into four parts. After 30 minutes of darkness, a light source was turned on, and flies’ positions were recorded to measure their light response. 2. Two-Vial Setup: Two connected vials were used, one covered in aluminum foil to create darkness. A light source illuminated the other vial, and flies’ positions in the light or dark vial were recorded. light but dispersed randomly after a few minutes. In the two-vial setup, control flies mostly chose the illuminated vial, whereas fewer hypoxia-treated flies did so, further highlighting sensory deficits. Conclusion: These results indicate that flies exposed to hypoxia under well-controlled conditions show clear deficits across behavioral assays. Impairments in motor function were evident in the locomotion assay, while disruptions in sensory-cognitive processes, such as decision-making and memory, were observed in the chemotaxis and phototaxis assays. This suggests that ischemic injury affects both movement and higher-order neural functions, offering insights into the broader effects of brain ischemia.

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