Exploring the connection between autophagy and heat-stress tolerance in Drosophila melanogaster
Data files
Sep 25, 2023 version files 24.74 MB
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Immunofluoresence_data.zip
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README.md
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Survival_and_developemental_data.zip
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Western_blotting_data.pdf
Sep 25, 2023 version files 24.74 MB
-
Immunofluoresence_data.zip
-
README.md
-
Survival_and_developemental_data.zip
-
Western_blotting_data.pdf
Abstract
Mechanisms aimed at recovering from heat-induced damages are closely associated with the ability of ectotherms to survive exposure to stressful temperatures. Autophagy, a ubiquitous stress-responsive catabolic process, has recently gained renewed attention as one of these mechanisms. By increasing the turnover of cellular structures as well as the clearance of long-lived protein and protein aggregates, the induction of autophagy has been linked to increased tolerance to a range of abiotic stressors in diverse ectothermic organisms. However, whether a link between autophagy and heat-tolerance exists in insect models remains unclear despite broad ecophysiological implications thereof. Here, we explored the putative association between autophagy and heat-tolerance using Drosophila melanogaster as a model. We hypothesized that (i) heat-stress would cause an increase of autophagy in flies' tissues, and (ii) rapamycin exposure would trigger a detectable autophagic response in adults and increase their heat-tolerance. In line with our hypothesis, we report that flies exposed to heat-stress present signs of protein aggregation and appear to trigger an autophagy-related homoeostatic response as a result. We further show that rapamycin feeding causes the systemic effect associated with target of rapamycin (TOR) inhibition, induces autophagy locally in the fly gut, and increases the heat-stress tolerance of individuals. These results argue in favour of a substantial contribution of autophagy to the heat-stress tolerance mechanisms of insects.
README: Title of Dataset
Exploring the Connection between Autophagy and Heat-Stress Tolerance in Drosophila melanogaster
General description: files include
Files include:
(1) Survival and developemental data.zip:
- Developmental_assay.txt
- Survival_assays_37.txt
- Survival_assays_41.txt:
- Recovery_assays_41.txt
(2) Immunofluoresence data.zip:
- Control folder
- 50 μm rapamycin folder
- 200 μm rapamycin folder
- Confocal thresholding.xlsx
(3) Western blotting data.pdf
Date of data collection: 2019-2023
Localisation of collection: Stellenbsoch
Links to publications that cite or use the data:
Willot, Q., du Toit, A., de Wet, S., Huisamen, E. J., Loos, B., & Terblanche, J. S. (2023). Exploring the connection between autophagy and heat-stress tolerance in Drosophila melanogaster. Proceedings of the Royal Society B, 290, 20231305.
Description of the data and file structure
(1) Survival and developemental data.zip
- Developmental_assay.txt: Rate of pupation and hatching of D. melanogaster larvae with and without 50 μM rapamycin exposure in the Kaplan-Meier curve format.
- Survival_assays_37.txt: Survival of D. melanogaster flies to a static 37°C heat-stress in the Kaplan-Meier curve format in either control conditions or exposed to 50 and 200 μM of rapamycin.
- Survival_assays_41.txt: Survival of D. melanogaster flies to a static 41°C heat-stress in the Kaplan-Meier curve format in either control conditions or exposed to 50 and 200 μM of rapamycin.
- Recovery_assays_41.txt: recovery rate of D. melanogster flies from a static 41°C heat-stress in the Kaplan-Meier curve format in either control conditions or exposed to 50 and 200 μM of rapamycin.
(2) Immunofluoresence data.zip
- Control folder: Portions and areas of the Drosophila midgut stained with lysotracker and imaged without rapamycin exposure. Each image replicate contains the raw image (.tif) and the processed image (8-bit greyscale) in ImageJ (counted area and pixels above lower set intensity threshold).
- 50 μm rapamycin folder: Portions and areas of the Drosophila midgut stained with lysotracker and imaged with 50 μm exposure. Each image replicate contains the raw image (.tif) and the processed image (8-bit greyscale) in ImageJ (counted area and pixels above lower set intensity threshold)
- 200 μm rapamycin folder: Portions and areas of the Drosophila midgut stained with lysotracker and imaged with 200 μm rapamycin exposure.Each image replicate contains the raw image (.tif) and the processed image (8-bit greyscale) in ImageJ (counted area and pixels above lower set intensity threshold)
- Confocal thresholding.xlsx: details for quantification of fluroresent pixels above set threshold for each processed image of each replicate of each condition
(3) Western blotting data.pdf: western blotting in triplicate against GABARAP proteins, Ref(2)P, and HSP70 in whole Drosophila tissue extracts following either rapamycin exposure and/or 1h heat-stress at 37°C. N= replicicate. Signal is given for both protein of interest and total protein signal recorded for standardization.
Sharing/access information
1. Links to other publicly accessible locations of the data: https://doi:10.5061/dryad.4qrfj6qf9
2. Was data derived from another source? No
A. If yes, list source(s): NA
3. Recommended citation for this dataset:
Willot, Q., du Toit, A., de Wet, S., Huisamen, E. J., Loos, B., & Terblanche, J. S. (2023). Exploring the connection between autophagy and heat-stress tolerance in Drosophila melanogaster. Proceedings of the Royal Society B, 290, 20231305.
Methods
Confocal Immunofluoresence Imagery of lysotracker signal in the Drosophila gut
Western blotting against autophagy markers
Survival assays of Drosophila flies