DOI: 10.5061/dryad.4qrfj6qf9
title: Exploring the connection between autophagy and heat-stress tolerance in Drosophila melanogaster

description: 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.
description: Confocal Immunofluoresence Imagery of lysotracker signal in the
 <em>Drosophila</em> gut<br>Western blotting against
 autophagy markers<br>Survival assays of
 <em>Drosophila</em> flies
description: # 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.