Endurance Training Protocol and Longitudinal Performance Assays for &lt;em&gt;Drosophila melanogaster&lt;/em&gt;

Tinkerhess, Martin J.; Ginzberg, Sara; Piazza, Nicole; Wessells, Robert J.

doi:10.3791/3786-v

Cited by 8 publications

(6 citation statements)

References 4 publications

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“…We found that endurance at day 5, measured as time-to-fatigue and plotted as a survival curve (runspan), was already significantly reduced in RU + adult males (On, red lines) compared to their RU-control (Off, black lines, Figure 4B). Then, upon completing the training program, the exercised-trained flies (EX) experienced a series of adaptations, including increased endurance and increased climbing speed compared to age-matched controls for untreated flies (Off, Figure 4 C-D, EX, solid black line vs UN, dashed black line) (Tinkerhess et al . 2012; Sujkowski and Wessells 2018).…”

Section: Resultsmentioning

confidence: 99%

“…). Then, upon completing the training program, the exercised-trained flies (EX) experienced a series of adaptations, including increased endurance and increased climbing speed compared to age-matched controls for untreated flies (Off, Figure4C-D, EX, solid black line vs UN, dashed black line)(Tinkerhess et al 2012;Sujkowski and Wessells 2018). However, turning On the GS system with MHC-GS-GAL4 reduced the climbing endurance after three weeks of exercise training (runspan), as well as the acute climbing speed (Figure4C-D, On EX, solid red line vs Off EX, solid black line).…”

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confidence: 99%

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The RU486-dependent activation of the GeneSwitch system in adult muscles leads to severe adverse effects in Drosophila

Zappia,

Damschroder,

Westacott

et al. 2023

Preprint

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Robust genetic systems to control the expression of transgenes in a spatial and temporal manner are a valuable asset for researchers. The GeneSwitch system induced by the drug RU486 has gained widespread use in the Drosophila community. However, some concerns were raised as negative effects were seen depending on the stock, transgene, stage and tissue under study. Here, we characterized the adverse effects triggered by activating the GeneSwitch system in adult muscles using theMHC-GS-GAL4driver. When a control, mockUAS-RNAitransgene was induced by feeding adult flies with RU486, we found that the overall muscle structure, including myofibrils and mitochondrial shape, was significantly disrupted and led to a significant reduction in the lifespan. Remarkably, lifespan was even shorter when two copies of the driver were used even without the mockUAS-RNAitransgene. Thus, researchers should be cautious when interpreting the results given the adverse effects we found when inducing RU486-dependentMHC-GS-GAL4in adult muscles. To counter the impact of these effects we recommend setting up additional control groups, such as a mock UAS-RNAi transgene, to validate the findings when using this inducible genetic system, as comparing the phenotypes between RU486-treated and untreated animals could be insufficient.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

The RU486-dependent activation of the GeneSwitch system in adult muscles leads to severe adverse effects in Drosophila

Zappia,

Damschroder,

Westacott

et al. 2023

Preprint

Self Cite

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“…Fatigue was recognized as the failure that responded to a negative geotaxis stimulus with climbing behavior. The specific measurement protocol is referred to the study of Tinkerhess et al 17 …”

Section: Methodsmentioning

confidence: 99%

“…16 When constructing the exercise device, the advantage of the flies' natural negative geotaxis behavior was taken to induce upward walking. 17 When flies climbed and reached the top of vial, the top of vial would be converted to the bottom by rotating the vial. 18,19 Flies were exercised in vial with an 8-cm length.…”

Section: Exercise Training Protocolsmentioning

confidence: 99%

Physical exercise ameliorates age‐related deterioration of skeletal muscle and mortality by activating Pten‐related pathways in Drosophila on a high‐salt diet

Hou,

Wen,

Zhong

et al. 2023

The FASEB Journal

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The phosphatase and tensin congeners (Pten) gene affects cell growth, cell proliferation, and rearrangement of connections, and it is closely related to cellular senescence, but it remains unclear the role of muscle‐Pten gene in exercise against age‐related deterioration in skeletal muscle and mortality induced by a high‐salt diet (HSD). In here, overexpression and knockdown of muscle Pten gene were constructed by building MhcGAL4/PtenUAS‐overexpression and MhcGAL4/PtenUAS‐RNAi system in flies, and flies were given exercise training and a HSD for 2 weeks. The results showed that muscle Pten knockdown significantly reduced the climbing speed, climbing endurance, GPX activity, and the expression of Pten, Sirt1, PGC‐1α genes, and it significantly increased the expression of Akt and ROS level, and impaired myofibril and mitochondria of aged skeletal muscle. Pten knockdown prevented exercise from countering the HSD‐induced age‐related deterioration of skeletal muscle. Pten overexpression has the opposite effect on skeletal muscle aging when compared to it knockdown, and it promoted exercise against HSD‐induced age‐related deterioration of skeletal muscle. Pten overexpression significantly increased lifespan, but its knockdown significantly decreased lifespan of flies. Thus, current results confirmed that differential expression of muscle Pten gene played an important role in regulating skeletal muscle aging and lifespan, and it also affected the adaptability of aging skeletal muscle to physical exercise since it determined the activity of muscle Pten/Akt pathway and Pten/Sirt1/PGC‐1α pathway.

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“…According to the Drosophila against gravity climbing characteristics, Drosophila endurance exercise training, using Drosophila endurance training exercise device, its principle is when the bottom of the test tube Drosophila climbs to the top of the test tube, the test tube begins to turn at a constant speed of 60 rad/s, until the top and the bottom exchange, at this time Drosophila climb to the top again, so cycle. [14][15][16] Reserve 8 cm of culture medium from the bottom of the test tube to the bottom of the cotton plug for the Drosophila movement area. After the two ends of the test tube change directions, there are 10 s for climbing.…”

Section: Exercise Trainingmentioning

confidence: 99%

Muscle‐specific overexpression of Atg2 gene and endurance exercise delay age‐related deteriorations of skeletal muscle and heart function via activating the AMPK/Sirt1/PGC‐1α pathway in male Drosophila

Wang,

Wen,

Wang

et al. 2023

The FASEB Journal

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Atg2 is a key gene in autophagy formation and plays an important role in regulating aging progress. Exercise is an important tool to resist oxidative stress in cells and delay muscle aging. However, the relationship between exercise and the muscle Atg2 gene in regulating skeletal muscle aging remains unclear. Here, overexpression or knockdown of muscle Atg2 gene was achieved by constructing the AtgUAS/MhcGal4 system in Drosophila, and these flies were also subjected to an exercise intervention for 2 weeks. The results showed that both overexpression of Atg2 and exercise significantly increased the climbing speed, climbing endurance, cardiac function, and lifespan of aging flies. They also significantly up‐regulated the expression of muscle Atg2, AMPK, Sirt1, and PGC‐1α genes, and they significantly reduced muscle malondialdehyde and triglyceride. These positive benefits were even more pronounced when the two were combined. However, the effects of Atg2 knockdown on skeletal muscle, heart, and lifespan were reversed compared to its overexpression. Importantly, exercise ameliorated age‐related changes induced by Atg2 knockdown. Therefore, current results confirmed that both overexpression of muscle Atg2 and exercise delayed age‐related deteriorations of skeletal muscle, the heart function, and lifespan, and exercise could also reverse age‐related changes induced by Atg2 knockdown. The molecular mechanism is related to the overexpression of the Atg2 gene and exercise, which increase the activity of the AMPK/Sirt1/PGC‐1α pathway, oxidation and antioxidant balance, and lipid metabolism in aging muscle.

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Endurance Training Protocol and Longitudinal Performance Assays for <em>Drosophila melanogaster</em>

Cited by 8 publications

References 4 publications

The RU486-dependent activation of the GeneSwitch system in adult muscles leads to severe adverse effects in Drosophila

The RU486-dependent activation of the GeneSwitch system in adult muscles leads to severe adverse effects in Drosophila

Physical exercise ameliorates age‐related deterioration of skeletal muscle and mortality by activating Pten‐related pathways in Drosophila on a high‐salt diet

Muscle‐specific overexpression of Atg2 gene and endurance exercise delay age‐related deteriorations of skeletal muscle and heart function via activating the AMPK/Sirt1/PGC‐1α pathway in male Drosophila

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