DOI: 10.5061/dryad.bcc2fqzjn
title: Experiencing short heat waves early in development changes thermal responsiveness of turtle embryos to later heat waves

description: Although physiological responses to the thermal environment are most
 frequently investigated using constant temperatures, the incorporation of
 thermal variability can allow for a more accurate prediction of how
 thermally sensitive species respond to a rapidly changing climate. In
 species with temperature-dependent sex determination (TSD), developmental
 responses to incubation temperature are mediated by several genes involved
 in gonadal differentiation. Kdm6b and Dmrt1 respond to cool incubation
 temperatures and are associated with testis development, while Foxl2 and
 Cyp19A1 respond to warm incubation temperatures and are associated with
 ovary development. Using fluctuating incubation temperatures, we designed
 two studies, one investigating how conflicting thermal cues affect the
 timing of commitment to gonadal development, and another investigating the
 rapid molecular responses to conflicting thermal cues in the red-eared
 slider turtle (Trachemys scripta). Using gene expression as a proxy of
 timing of commitment to gonadal fate, results from the first study show
 that exposure to high amounts of conflicting thermal cues during
 development delays commitment to gonadal fate. Results from the second
 study show that Kdm6b splice variants exhibit differential responses to
 early heat wave exposure, but rapidly (within 2 days) recover to
 pre-exposure levels after the heat wave. Despite changes in the expression
 of Kdm6b splice variants, there was no effect on Dmrt1 expression.
 Collectively, these findings demonstrate how short exposures to heat early
 in development can change how embryos respond to heat later in
 development.
description: # Experiencing short heat waves early in development changes thermal
 responsiveness of turtle embryos to later heat waves ## Description of the
 Data and file structure Each data file (one for each of the two studies)
 has gene expression values obtained from biological samples from multiple
 genes. Each data file also has information on the treatment and sampling
 point of each individual analysed in both studies. Rapid response study
 data column headers: * Egg number: identifies individual T. scripta
 embryos used in study * Experiment: shows which gene is being analyzed *
 Treatment: shows whether or not individuals were exposed to heat wave *
 Stage: shows which embryonic stage individual was sampled at * Day: shows
 day of incubation that individual was sampled on * Gapdh: expression (CT)
 for house keeping gene * Dmrt1: expression of Dmrt1 gene * Kdm6bGe:
 expression of Kdm6b general (both splicing variants) * Kdm6bUp3:
 expression of Kdm6b(-IR) * Kdm6b(+IR): expression of Kdm6b(+IR) * DeltaCT:
 expression of gene of interest minus expression of housekeeping gene *
 Normalized: normalized expression following delta delta CT method * Log:
 log of normalized expression Delayed response study column headers: *
 Tube: identifies sample tube that cDNA for each sample is stored in *
 Sample: identifies the egg number of sampled embryo * TRT: shows
 temperature treatment * Day: shows day of incubation embryo was sampled *
 Stage: shows stage of development individual was sampled * Mean Gapdh CT:
 mean expression for housekeeping gene * NDmrt1: normalized Dmrt1
 expression * NCyp19A1: normalized Cyp19A1 expression * NFoxl2: normalized
 Foxl2 expression * NSox9: normalized Sox9 expression * NKdm6b: normalized
 Kdm6b(+IR) expression Each of the R script codes show how we analyzed our
 data for both experiments and how assumptions of GLMs were checked, as
 well as how post-hoc comparisons were carried out and how they were
 adjusted. ## Sharing/access Information NA