FLEX: Flight Exercise Training Protocol for the Fruit Fly Drosophila

Our research focuses on transgenerational susceptibility to neurological and metabolic disorders. We investigate molecular mechanisms, specifically bioenergetic changes that might transmit susceptibility across generations. Omics techniques like transcriptomics, proteomics, and metabolomics can help identify the mechanisms and causes of neurodegenerative disorders.

To begin, keep the Drosophila flies in an incubator or room maintained at 24 degrees Celsius with 60%humidity and a 12-hour light and dark cycle. Collect age-matched flies within three to four days after eclosion. Randomly assign the flies to exercise and sedentary groups.

Provide both groups with seven milliliters of formulation food and use tape to secure the food vials inside the bowls. If humidity cannot be controlled, provide water access by placing vials of wet cotton inside the bowls and taping them to the bowl walls. Transfer flies into the bowls one day before the exercise regimen begins.

Cover the openings of the bowls with mesh and secure it using rubber bands. Conduct the training in a room maintained at 24 degrees Celsius with 60%humidity. Secure the bowls to the exercise platform using bungee cords.

Use two timers to control the motor. Set the first timer to start the machine at 8:00 a.m. and stop it at 3:00 p.m.daily.

Program the second timer to trigger a series of three motor revolutions every five minutes. Each revolution should raise and drop the platform, inducing flight in the flies. At the end of the FLEX regimen, place the bowls in a cold room for five minutes to immobilize the flies.

Then, transfer the flies into new food vials for phenotypic analysis or into micro centrifuge tubes for molecular or biochemical assays. The five-day flight exercise regimen significantly reduced mortality in flies on a Western diet. Flight exercise also improved climbing ability in Western-diet-fed flies.

Maximal oxygen consumption during complex-one-supported respiration was reduced by 42%in flight muscles from Western-diet-fed flies compared to controls, but was restored in flies from the Western diet plus exercise group. Offspring of Western-diet-fed fathers showed significantly more feeding licks than other groups, while paternal exercise negated this increase. We found that paternal Western diet reprograms offspring phenotype, including changes in activity, learning and memory, and feeding behavior.

Many protocols depend on forced climbing behavior. Flight exercise is more physiological and doesn't lead to injuries. This protocol will utilize more physiological exercise that generates significant bioenergetic demands and minimize injuries.