DOI: 10.5061/dryad.gf1vhhmxm
title: Data from: HIF-signaling in the prothoracic gland regulates growth and development in hypoxia but not normoxia in Drosophila

description: The developmental regulation of body size is a fundamental life-history
 characteristic that in most animals is tied to the transition from
 juvenile to adult form. In holometabolous insects this transition is
 ostensibly initiated at the attainment of a critical weight in the final
 larval instar. It has been hypothesized that the size-sensing mechanism
 used to determine attainment of critical weight exploits oxygen limitation
 as a larvae grows beyond the oxygen-delivery capacity of its fixed
 tracheal system; that is, developmentally-induced cellular hypoxia
 initiates the synthesis of the molting hormone ecdysone by the prothoracic
 gland. We tested this hypothesis in Drosophila by assaying cellular
 hypoxia throughout the third-larval instar at 21 and 10 kPa O2, using the
 activity of the HIF-signalling pathway as a measure of hypoxia. While
 HIF-signalling was elevated at low levels of environmental O2 it did not
 markedly increase during development at either oxygen level, and was only
 suppressed by hyperoxia after feeding had ceased. Further, changes in
 HIF-signalling in the prothoracic gland alone did not alter body size or
 developmental time in a way that would be expected if cellular hypoxia in
 the prothoracic gland was part of the critical weight mechanism. Our data
 do show, however, that reduced HIF-signalling in the prothoracic gland
 decreases survival and retards development at 10 kPa O2, suggesting that
 prothoracic HIF-signaling is a necessary part of the beneficial plasticity
 mechanism that controls growth and development in response to low oxygen
 level.
description: The data include: Gene expression
 analysis of HIF-signaling genes in <em>Drosophila
 melanogaster</em> third-instar larvae reared at 10, 21 and 40 kPa
 O<sub>2</sub>. Western blot analysis of
 HIF-1α protein in <em>Drosophila melanogaster</em>
 third-instar larvae reared at 10, 21 and 40 kPa
 O<sub>2</sub>. Adult body size,
 developmental time, and survival of <em>Drosophila melanogaster
 r</em>eared at 10 and 21 kPa O<sub>2,</sub> in which
 HIF-signaling in the prothoracic gland has been up- and
 down-regulated.
description: # Data from: HIF-signaling in the prothoracic gland regulates growth and
 development in hypoxia but not normoxia in Drosophila
 [https://doi.org/10.5061/dryad.gf1vhhmxm](https://doi.org/10.5061/dryad.gf1vhhmxm) ## Description of the data and file structure The data were collected from *Drosophila* larvae and adults reared in hypoxia (10kPa O2), normoxia (21kPa O2), or hyperoxia (40kPa O2).  *Drosophila* were either wildtype controls or had up- and down-regulated HIF signaling in the PG by knockdown of *Hph* and *HIF-1α *expression, respectively, using phm-GAL4 to drive expression of *UAS*-*Hph*.RNAi and *UAS*-*HIF-1α.RNAi*. ### Files and variables The archive comprises six files. An RMarkdown file (Analysis.rmd) of the annotated code used to conduct the analysis and five text files (.csv) of the data used in the analysis. The data files are as follows: 1. Expression.csv contains the gene expression data. Variables include which gene is being analyzed (gene), experimental treatment (treatment), age in hours from ecdysis to third instar as a categorical variable (age), age in hours from ecdysis to third instar as a continuous variable (agenum), copy number (copies), pooled age as a categorical variable (pooled), pooled age as a continuous variable (poolnum), oxygen level in percent as a categorical variable (ox), and oxygen level in percent as a continuous variable (oxnum). 2. Westen.csv contains the western blot data assaying HIF-1α protein levels. Variables include age of larvae (time), oxygen level in precent (oxygen), and protein level (density and logdensity, unitless). 3. Body Size.csv contains the body size data. Variables include cross (cross: fga/phm – knockdown of *Hph*; sima/phm – knockdown of *HIF-1α*), oxygen level in percent (o2), sex (sex), genotype (genotype: +/+, uas/+, gal4/+, uas/gal4), adult mass in mg (mass), and experimental type (type: control, exp). 4. Developmental Time.csv contains the developmental time data. Variables include cross (uas: fga [*Hph*], sima [*HIF-1α*]), oxygen level in percent (o2), genotype (genotype: +/+, fga/+, sima/+, phm/+, fga/phm, sima/phm), total number of flies (total.flies), age in hours from oviposition (age), number of eclosed flies at each age (eclose), number of uneclosed flies at each age (not.eclose), and experimental type (type: control, exp).  5. Survival.csv contains the survival data. Variables include cross (uas: fga [*Hph*], sima [*HIF-1α*]), oxygen level in percent (o2), total number of flies (total.flies), genotype (genotype: +/+, uas/+, gal4/+, uas/gal4), number of flies that eclose as adults (eclose), number of flies that do not eclose as adults (not.eclose), and experimental type (type: control, exp).  ## Code/software The analysis can be conducted in R using RStudio (2022.07.1) and R (4.0.3). The R Markdown file contains the annotated script of all the analysis described in the paper.