Name: FLEX: Flight Exercise Training Protocol for the Fruit Fly Drosophila (Video) | JoVE Journal
Uploaded: 2025-10-14T14:20:00.000+00:00
Duration: 3 min 47 s
Description: Learn flight exercise training for Drosophila to enhance aerobic capacity and metabolic health. This protocol helps researchers study transgenerational effects of exercise on neurological and metabolic disorders.

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5 minutes

This research was supported by NIDDK R01DK129455 (AKM).

1. Exercise machine setup and operation
<ol>
	<li>Build a machine at the University Machine Shop. Use a 35 mm x 35 mm x 1.4 mm rectangular aluminum base with an AC/DC motor and a 35 mm x 35 mm x 0.5 mm horizontal aluminum platform (Figure 1A).</li>
	<li>For the conversion of rotary motion to linear motion, build a custom-made rotating round cam 15 cm in diameter mounted on the motor (Figure 1B). Attach a follower with a 2 cm roller to the bottom of the horizontal platform so that the platform is lifted by the follower and then dropped back to its original position once the follower roller clears the cam.</li>
	<li>Adjust the speed of the motor with a rotary potentiometer such that a full rotational period is about 5,360 ms long. 
	NOTE: Exact motor specifications, dimensions of follower and roller, as well as cam geometry, are not critical. Any motorized configuration that reliably lifts and drops the platform to dislodge the flies and permits user control over the drop frequency is acceptable.</li>
	<li>Secure 1-gallon clear plastic drum fish bowls to the platform by bungee cords that are hooked onto screw eyes (Figure 1C).</li>
</ol>
2. FLEX protocol
<ol>
	<li>Keep flies in a 24 &#176;C incubator or room with 60% humidity and a 12 h light/dark cycle.</li>
	<li>Collect age-matched flies within 3-4 days of hatching. After collection, randomly assign flies to exercise and sedentary groups. Use an equal number of flies for both the exercise and sedentary groups. 
	NOTE: Flies may be housed at densities ranging from 30 to 400 per bowl, as this does not interfere with flight exercise. However, experimental and control groups should always be matched for number, sex, and age to ensure comparability.</li>
	<li>Provide sedentary and controlled flies with 7 mL of formulation food. Secure food vials in the bowls with tape.</li>
	<li>Transfer flies into the bowls using light and brief CO2 anesthesia a day before starting the exercise regimen. Cover bowls&#39; openings with a mesh secured with rubber bands. 
	NOTE: Flies should be anesthetized briefly under light CO2 on a pad. The exposure must remain short to avoid lasting behavioral effects; therefore, anesthesia should be performed 1 day before the exercise experiment.</li>
	<li>Secure the bowls to the platform with bungee cords. Conduct the exercise training in a room with a temperature of 24 &#176;C and a humidity of 60%. If the humidity in the room is not controlled, provide flies with access to water by placing vials of wet cotton into the fish bowls and securing vials to the bowl&#39;s wall with tape.</li>
	<li>For control experiments (sham exercise), place an equal number of flies in a bowl positioned next to the exercise machine but not attached to the platform.</li>
	<li>Exercise flies for 5 consecutive days.
	<ol>
		<li>Control the motor with two timers (Figure 1D). The first timer turns the machine on and off at a specific time of the day. Begin exercise at 8 am and finish at 3 pm.</li>
		<li>Use the second timer to initiate a series of three motor revolutions, with each sequence occurring every 5 min. Each revolution will elevate the platform and then drop it down, triggering the flies into flight.</li>
		<li>Once the exercise treatment for a given day is completed, return the bowls to their incubators. This protocol can be easily modified to suit a particular experiment. 
		NOTE: A humidity below 60% may cause an increase in mortality. A small number of flies may not initiate flight after the first drop, but typically begin flying after the second or third drop. While it is not feasible to track flight in every individual, the majority of flies engage in flight, and it is assumed that all flies eventually participate in exercise. In addition, the large container provides space for voluntary flight between drops.</li>
	</ol>
	</li>
	<li>At the end of the FLEX regimen, immobilize flies by placing the bowls in the cold room for 5 min. Then transfer flies to new food vials for further analyses of phenotypic characteristics or to microcentrifuge tubes for molecular and/or biochemical assay of interest. 
	NOTE: Examples of further analyses include behavioral and biochemical assay and/or gene expression. Assays involving living flies will need to be conducted promptly in accordance with the specific goals of the phenotypic assay. The protocol can be paused here with samples for biochemistry/molecular biology frozen and stored appropriately, depending on the assessments to be performed. A summary of the protocol is provided as Table 1.</li>
</ol>

<ol><li>Ruegsegger, G. N., Booth, F. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Health+benefits+of+exercise%5BTitle%5D)+AND+%22Cold+Spring+Harb+Perspect+Med%22%5BJournal%5D)">Health benefits of exercise.</a> Cold Spring Harb Perspect Med. 8 (7), a029694(2018).</li>
<li>Valenzuela, P. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Exercise+benefits+on+Alzheimer%27s+disease%3A+State-of-the-science%5BTitle%5D)+AND+%22Ageing+Res+Rev%22%5BJournal%5D)">Exercise benefits on Alzheimer's disease: State-of-the-science.</a> Ageing Res Rev. 62, 101108(2020).</li>
<li>Petridou, A., Siopi, A., Mougios, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Exercise+in+the+management+of+obesity%5BTitle%5D)+AND+%22Metabolism%22%5BJournal%5D)">Exercise in the management of obesity.</a> Metabolism. 92, 163-169 (2019).</li>
<li>Fiuza-Luces, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Exercise+benefits+in+cardiovascular+disease%3A+Beyond+attenuation+of+traditional+risk+factors%5BTitle%5D)+AND+%22Nat+Rev+Cardiol%22%5BJournal%5D)">Exercise benefits in cardiovascular disease: Beyond attenuation of traditional risk factors.</a> Nat Rev Cardiol. 15 (12), 731-743 (2018).</li>
<li>Neufer, P. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Understanding+the+cellular+and+molecular+mechanisms+of+physical+activity-induced+health+benefits%5BTitle%5D)+AND+%22Cell+Metab%22%5BJournal%5D)">Understanding the cellular and molecular mechanisms of physical activity-induced health benefits.</a> Cell Metab. 22 (1), 4-11 (2015).</li>
<li>Hastings, M. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Animal+models+of+exercise+from+rodents+to+pythons%5BTitle%5D)+AND+%22Circ+Res%22%5BJournal%5D)">Animal models of exercise from rodents to pythons.</a> Circ Res. 130 (12), 1994-2014 (2022).</li>
<li>Garton, F. C., North, K. N., Koch, L. G., Britton, S. L., Nogales-Gadea, G., Lucia, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Rodent+models+for+resolving+extremes+of+exercise+and+health%5BTitle%5D)+AND+%22Physiol+Genomics%22%5BJournal%5D)">Rodent models for resolving extremes of exercise and health.</a> Physiol Genomics. 48 (2), 82-92 (2016).</li>
<li>Sujkowski, A., Wessells, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Using+Drosophila+to+understand+biochemical+and+behavioral+responses+to+exercise%5BTitle%5D)+AND+%22Exerc+Sport+Sci+Rev%22%5BJournal%5D)">Using Drosophila to understand biochemical and behavioral responses to exercise.</a> Exerc Sport Sci Rev. 46 (2), 112-120 (2018).</li>
<li>Watanabe, L. P., Riddle, N. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((New+opportunities%3A+Drosophila+as+a+model+system+for+exercise+research%5BTitle%5D)+AND+%22J+Appl+Physiol%22%5BJournal%5D)">New opportunities: Drosophila as a model system for exercise research.</a> J Appl Physiol. 127 (2), 482-490 (2019).</li>
<li>Ding, M., Li, H., Zheng, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Drosophila+exercise%2C+an+emerging+model+bridging+the+fields+of+exercise+and+aging+in+human%5BTitle%5D)+AND+%22Front+Cell+Dev+Biol%22%5BJournal%5D)">Drosophila exercise, an emerging model bridging the fields of exercise and aging in human.</a> Front Cell Dev Biol. 10, 966531(2022).</li>
<li>Tinkerhess, M. J., Ginzberg, S., Piazza, N., Wessells, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Endurance+training+protocol+and+longitudinal+performance+assays+for+Drosophila+melanogaster%5BTitle%5D)+AND+%22J+Vis+Exp%22%5BJournal%5D)">Endurance training protocol and longitudinal performance assays for Drosophila melanogaster.</a> J Vis Exp. (61), e3786(2012).</li>
<li>Watanabe, L. P., Riddle, N. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Characterization+of+the+rotating+exercise+quantification+system+%28REQS%29%2C+a+novel+Drosophila+exercise+quantification+apparatus%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Characterization of the rotating exercise quantification system (REQS), a novel Drosophila exercise quantification apparatus.</a> PLoS One. 12 (10), e0185090(2017).</li>
<li>Piazza, N., Gosangi, B., Devilla, S., Arking, R., Wessells, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Exercise-training+in+young+Drosophila+melanogaster+reduces+age-related+decline+in+mobility+and+cardiac+performance%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Exercise-training in young Drosophila melanogaster reduces age-related decline in mobility and cardiac performance.</a> PLoS One. 4 (6), e5886(2009).</li>
<li>Lowman, K. E., Wyatt, B. J., Cunneely, O. P., Reed, L. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+TreadWheel%3A+Interval+training+protocol+for+gently+induced+exercise+in+Drosophila+melanogaster%5BTitle%5D)+AND+%22J+Vis+Exp%22%5BJournal%5D)">The TreadWheel: Interval training protocol for gently induced exercise in Drosophila melanogaster.</a> J Vis Exp. (136), e57788(2018).</li>
<li>Cheng, X., Sun, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Wing-kinematics+measurement+and+aerodynamics+in+a+small+insect+in+hovering+flight%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">Wing-kinematics measurement and aerodynamics in a small insect in hovering flight.</a> Sci Rep. 6, 25706(2016).</li>
<li>Aono, H., Liang, F., Liu, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Near-+and+far-field+aerodynamics+in+insect+hovering+flight%3A+An+integrated+computational+study%5BTitle%5D)+AND+%22J+Exp+Biol%22%5BJournal%5D)">Near- and far-field aerodynamics in insect hovering flight: An integrated computational study.</a> J Exp Biol. 211 (Pt 2), 239-257 (2008).</li>
<li>Bennett, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Insect+aerodynamics%3A+Vertical+sustaining+force+in+near-hovering+flight%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">Insect aerodynamics: Vertical sustaining force in near-hovering flight.</a> Science. 152 (3726), 1263-1266 (1966).</li>
<li>Fry, S. N., Sayaman, R., Dickinson, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+aerodynamics+of+hovering+flight+in+Drosophila%5BTitle%5D)+AND+%22J+Exp+Biol%22%5BJournal%5D)">The aerodynamics of hovering flight in Drosophila.</a> J Exp Biol. 208 (Pt 12), 2303-2318 (2005).</li>
<li>Lane, S. J., Frankino, W. A., Elekonich, M. M., Roberts, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+effects+of+age+and+lifetime+flight+behavior+on+flight+capacity+in+Drosophila+melanogaster%5BTitle%5D)+AND+%22J+Exp+Biol%22%5BJournal%5D)">The effects of age and lifetime flight behavior on flight capacity in Drosophila melanogaster.</a> J Exp Biol. 217 (Pt 9), 1437-1443 (2014).</li>
<li>Margotta, J. W., Roberts, S. P., Elekonich, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effects+of+flight+activity+and+age+on+oxidative+damage+in+the+honey+bee%2C+Apis+mellifera%5BTitle%5D)+AND+%22J+Exp+Biol%22%5BJournal%5D)">Effects of flight activity and age on oxidative damage in the honey bee, Apis mellifera.</a> J Exp Biol. 221 (Pt 14), jeb183228(2018).</li>
<li>Bartholomew, N. R., Burdett, J. M., VandenBrooks, J. M., Quinlan, M. C., Call, G. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Impaired+climbing+and+flight+behaviour+in+Drosophila+melanogaster+following+carbon+dioxide+anaesthesia%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">Impaired climbing and flight behaviour in Drosophila melanogaster following carbon dioxide anaesthesia.</a> Sci Rep. 5, 15298(2015).</li>
<li>Chambers, R. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Nicotine+increases+lifespan+and+rescues+olfactory+and+motor+deficits+in+a+Drosophila+model+of+Parkinson%27s+disease%5BTitle%5D)+AND+%22Behav+Brain+Res%22%5BJournal%5D)">Nicotine increases lifespan and rescues olfactory and motor deficits in a Drosophila model of Parkinson's disease.</a> Behav Brain Res. 253, 95-102 (2013).</li>
<li>Pesah, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Drosophila+parkin+mutants+have+decreased+mass+and+cell+size+and+increased+sensitivity+to+oxygen+radical+stress%5BTitle%5D)+AND+%22Development%22%5BJournal%5D)">Drosophila parkin mutants have decreased mass and cell size and increased sensitivity to oxygen radical stress.</a> Development. 131 (9), 2183-2194 (2004).</li>
<li>Murashov, A. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Preference+and+detrimental+effects+of+high+fat%2C+sugar%2C+and+salt+diet+in+wild-caught+Drosophila+simulans+are+reversed+by+flight+exercise%5BTitle%5D)+AND+%22FASEB+Bioadv%22%5BJournal%5D)">Preference and detrimental effects of high fat, sugar, and salt diet in wild-caught Drosophila simulans are reversed by flight exercise.</a> FASEB Bioadv. 3 (1), 49-64 (2021).</li>
<li>Murashov, A. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Paternal+Western+diet+causes+transgenerational+increase+in+food+consumption+in+Drosophila+with+parallel+alterations+in+the+offspring+brain+proteome+and+microRNAs%5BTitle%5D)+AND+%22FASEB+J%22%5BJournal%5D)">Paternal Western diet causes transgenerational increase in food consumption in Drosophila with parallel alterations in the offspring brain proteome and microRNAs.</a> FASEB J. 37 (6), e22966(2023).</li>
<li>Van der Horst, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Insect+adipokinetic+hormones%3A+Release+and+integration+of+flight+energy+metabolism%5BTitle%5D)+AND+%22Comp+Biochem+Physiol+B+Biochem+Mol+Biol%22%5BJournal%5D)">Insect adipokinetic hormones: Release and integration of flight energy metabolism.</a> Comp Biochem Physiol B Biochem Mol Biol. 136 (2), 217-226 (2003).</li>
<li>Suarez, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Energy+metabolism+during+insect+flight%3A+Biochemical+design+and+physiological+performance%5BTitle%5D)+AND+%22Physiol+Biochem+Zool%22%5BJournal%5D)">Energy metabolism during insect flight: Biochemical design and physiological performance.</a> Physiol Biochem Zool. 73 (6), 765-771 (2000).</li>
<li>Lehmann, F. O., Skandalis, D. A., Berthe, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Calcium+signalling+indicates+bilateral+power+balancing+in+the+Drosophila+flight+muscle+during+manoeuvring+flight%5BTitle%5D)+AND+%22J+R+Soc+Interface%22%5BJournal%5D)">Calcium signalling indicates bilateral power balancing in the Drosophila flight muscle during manoeuvring flight.</a> J R Soc Interface. 10 (82), 20121050(2013).</li>
</ol>

The authors have nothing to disclose.

The FLEX protocol provides a novel and physiologically relevant method for inducing aerobic exercise in Drosophila melanogaster through flight. Unlike traditional paradigms such as the Power Tower and TreadWheel, which rely on negative geotaxis and mechanical agitation, FLEX harnesses the fly&#39;s innate flight behavior. This approach minimizes exercise-induced injuries and stress responses, making it a safer and more biologically appropriate model for exercise biology.
Insects rely ...

Physical activity and exercise are the focus of intense research for their immediate and long-term health benefits1. Due to its powerful systemic effect on metabolism, exercise is increasingly used to prevent and treat metabolic, cardiovascular, and neurological disorders2,3,4. Despite the increasing evidence of the numerous health benefits associated with regular exercise, the exact molecular mechanisms underpinning those benefits remain poorly understood5. To understand these mechanisms, the use of innovative and high-throughput animal models is essential. While rodent models are commonly used to investigate the impact of exercise on metabolic diseases, such studies require significant costs, regulatory oversight, and specialized equipment6,7.
Recently, Drosophila melanogaster has emerged as a new model for researching exercise biology (reviewed8,9,10). Equipped with a wide array of potent genetic tools, a short life cycle, extensive progeny, and economical maintenance requirements, the fruit fly presents a valuable resource for investigating genetic, epigenetic, and transgenerational mechanisms. When modeling exercise in flies, current methods typically rely on negative geotaxis, the behavioral reaction to climb upwards11,12.
One of the first automated exercise protocols utilized a special exercise apparatus named the Power Tower11. The Power Tower uses a motor unit that repeatedly lifts vials with animals and then drops them down to induce the negative geotaxis response11. Prolonged exercise in the Power Tower results in a significant reduction in age-related decline in mobility and cardiac performance13. Additionally, the Power Tower regimen can be stressful for flies, and excessive training intensity has been reported to cause injury or mortality11.
To address this issue, another exercise paradigm called Treadwheel was developed14. The Treadwheel also leverages the negative geotaxis response of Drosophila, but instead of lifting and dropping flies as in the Power Tower, it inverts vials with animals. Unlike the Power Tower, this method is gentler on flies and prevents exercise-related injuries and mortality9. However, the flies in the Treadwheel could habituate to the rotating motion, which might complicate the interpretation of the results12. To address this issue, a Locomotor Activity Monitor (LAM25H, Trikinetics) was added to measure activity during exercise, to quantify Drosophila activity, and account for habituation. Subsequently, the name of the system was updated to the Rotational Exercise Quantification System (REQS)12.
Interestingly, very few studies took advantage of the natural flight abilities of Drosophila to model exercise. The predominant focus of flight research has been on the aerodynamics and biomechanical aspects of hovering and maneuvering in the air15,16,17,18. Several studies have delved into the physiological and cellular mechanisms related to flight19,20. In these studies, flight ability was assessed via the flight assay21,22, where individual flies were released into a clear acrylic box and their ability to maintain steady elevation and flight was observed23.
In one particular study, researchers examined the effects of induced flight exercise by placing flies in bottles attached to a vortex-type shaker. The shaker gently shook the bottles at random intervals between 5 min and 10 min, using a programmable timer19. This study reported that induced flight behavior accelerated senescence and the age-related loss of antioxidant capacity in flight muscles19. Building on the previous research, our laboratory has developed a new exercise protocol called FLEX (flight exercise) by integrating an automated negative geotaxis system with housing flies in large containers24. For the FLEX training, flies were placed in 1-gallon clear plastic drum fish bowls strapped to a horizontal platform attached to a motor. The motor was controlled by a timer, which initiated motor revolutions every 5 min. Each revolution elevated the platform and then dropped it down, triggering the flies into flight. The FLEX was performed daily for 7 h for 5 days. No mortalities or injuries associated with exercise were observed24. Moreover, a 5-day FLEX regimen in flies on the Western Diet (WD) markedly negated the adverse effects of the WD, improving survival, climbing behavior, and mitochondrial respiration24. Together, this paper presents a novel exercise paradigm based on natural flight activity that does not cause injury. This is a high-throughput procedure that allows a large number of flies to be trained simultaneously to study the interaction between exercise, diet, drugs, aging, environment, and/or genetics. The protocol can be used to complement existing exercise methods to study the effects of physical activity on fly physiology and behavior.

<table><tbody><tr><td>17 x 100 mm 14 mL polypropylene culture tube&amp;nbsp;</td><td>VWR</td><td>&amp;nbsp;60818-689</td><td></td></tr><tr><td>1-gallon clear&lt;br/&gt; plastic drum fish bowls&amp;nbsp;</td><td>Petco</td><td>N/A</td><td>Available from Petco</td></tr><tr><td>8 Outlet Surge Protector with 7-Day Digital Timer</td><td>BN-LINK&amp;nbsp;</td><td>EGPH020028</td><td>Available from Amazon</td></tr><tr><td>AC/DC motor&amp;nbsp;</td><td>Dayton&amp;nbsp;</td><td>Model 12W368 with torque 80 IN-LBS</td><td></td></tr><tr><td>Bloomington Formulation diet&amp;nbsp;</td><td>Nutri-Fly</td><td>&amp;nbsp;66-112</td><td>Available from Genesee Scientific Inc., San Diego, CA</td></tr><tr><td>Kim-wipes</td><td>Fisher Scientific</td><td>06-666</td><td>Kimberly-Clark Professional 34120</td></tr><tr><td>Outlet Multifunctional Programmable Timer with Countdown and Infinite Short Cycle</td><td>NEARPOW&amp;nbsp;</td><td>SPM10188153502</td><td>Available from Amazon</td></tr></tbody></table>

flex: flight exercise training protocol for the fruit fly drosophila

FLEX: Flight Exercise Training Protocol for the Fruit Fly Drosophila

Drosophila has recently become a popular model for studying obesity and related disorders. While it's relatively easy to induce metabolic disorders in Drosophila using specific diets, creating an exercise model for the flies is more challenging. Existing methods, such as the Power Tower and Treadwheel, utilize shaking or rotating vials to induce the flies to climb, leveraging their negative geotaxis response. However, continuous climbing for long periods is unnatural for the flies and can lead to exercise-related injuries and mortalities. This paper introduces a new exercise protocol that leverages the fly's natural ability to fly, offering a more natural, physiological, and injury-free exercise approach. The experiments show that a 5-day flight exercise regimen increases the fly's bioenergetic capacity and counteracts the adverse effects of a Western diet (WD), improving climbing behavior and reducing WD-associated mortality. This protocol enables the simultaneous training of a large number of flies to study the interaction between exercise and various factors, including diet, drugs, aging, and genetics. Together, this method can complement existing methods for studying the effects of exercise on fly physiology and behavior.

The FLEX protocol was previously validated in a published work from our laboratory. In a study published, we showed that flight exercise mitigates the negative metabolic effects of a Western Diet (WD)24. This diet was made on the basis of the standard Nutri-Fly Bloomington diet with the following additions of 15% Nutiva USDA Certified Organic, non-GMO, Red Palm Oil, 15% Sucrose, and 0.1 M NaCl.
Three-to-four-day old adult male fruit flies were randomly assigned to each ...

The goal of this protocol is to provide flight exercise for the fruit fly Drosophila. Representative results show that flight exercise training results in an increased aerobic capacity and resistance to a high-fat diet.

Drosophila has recently become a popular model for studying obesity and related disorders. While it's relatively easy to induce metabolic disorders in ...

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.

flex-flight-exercise-training-protocol-for-the-fruit-fly-drosophila

Research

JoVE Journal

Developmental Biology

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A method for rearing Drosophila melanogaster under axenic and gnotobiotic conditions is presented. Fly embryos are dechorionated in sodium hypochlorite, transferred aseptically to sterile diet, and reared in closed containers. Inoculating diet and embryos with bacteria leads to gnotobiotic associations, and bacterial presence is confirmed by plating whole-body Drosophila homogenates.

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In this paper, we provide a detailed protocol for exposing species in the genus Drosophila to pollutants with the goal of studying the impact of exposure on a range of phenotypic outputs at different developmental stages and for more than one generation.

Experimental Protocol for Using Drosophila As an Invertebrate Model System for Toxicity Testing in the Laboratory

The Rotating Exercise Quantification System (REQS) can induce exercise in Drosophila melanogaster through rotation while simultaneously measuring the amount of activity performed by the animals. Here, we present a point-by-point protocol detailing how to measure activity levels of animals experiencing rotational exercise treatments using the REQS.

Measuring Exercise Levels in Drosophila melanogaster Using the Rotating Exercise Quantification System (REQS)

The TreadWheel uses rotational motion to gently induce exercise in adult Drosophila melanogaster by exploiting flies' innate, negative geotaxis. It allows for analysis of the interactions between exercise and factors, such as genotype, sex, and diet, and their effect on physiological and molecular assays to assess metabolic health.

The TreadWheel: Interval Training Protocol for Gently Induced Exercise in Drosophila melanogaster

Described here is the use of several homemade tools to transfer, chill, and kill adult Drosophila, as well as to clean glass culture vials and collect eggs. These tools are easy to make and are rather efficient in handling Drosophila.

Simple Homemade Tools to Handle Fruit Flies&#8212;Drosophila melanogaster

Learn flight exercise training for Drosophila to enhance aerobic capacity and metabolic health. This protocol helps researchers study transgenerational effects of exercise on neurological and metabolic disorders.

FLEX: Flight Exercise Training Protocol for the Fruit Fly Drosophila

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