DOI: 10.5061/dryad.f4qrfj724
title: Data and code for: Acute heat priming promotes short-term climate resilience of early life stages in a model sea anemone

description: Across diverse taxa, sublethal exposure to abiotic stressors early in life
 can lead to benefits such as increased stress tolerance upon repeat
 exposure. This phenomenon, known as hormetic priming, is largely
 unexplored in early life stages of marine invertebrates, which are
 increasingly threatened by anthropogenic climate change. To investigate
 this phenomenon, larvae of the sea anemone and model marine invertebrate
 Nematostella vectensis were exposed to control (18°C) or elevated (24°C,
 30°C, 35°C, or 39°C) temperatures for 1 hour at 3 days post-fertilization
 (DPF), followed by return to control temperatures (18°C). The animals were
 then assessed for growth, development, metabolic rates, and heat tolerance
 at 4, 7, and 11 DPF. Priming at intermediately elevated temperatures
 (24°C, 30°C, or 35°C) augmented growth and development compared to
 controls or priming at 39°C. Indeed, priming at 39°C hampered
 developmental progression, with around 40% of larvae still in the planula
 stage at 11 DPF, in contrast to 0% for all other groups. Total protein
 content, a proxy for biomass, and respiration rates were not significantly
 affected by priming, suggesting metabolic resilience. Heat tolerance was
 quantified with acute heat stress exposures, and was significantly higher
 for animals primed at intermediate temperatures (24°C, 30°C, or 35°C)
 compared to controls or those primed at 39°C at all time points. To
 investigate a possible molecular mechanism for observed changes in heat
 tolerance, the expression of heat shock protein 70 (HSP70) was quantified
 at 11 DPF. Expression of HSP70 significantly increased with increasing
 priming temperature, with the presence of a doublet band for larvae primed
 at 39°C, suggesting persistent negative effects of priming on protein
 homeostasis. Interestingly, primed larvae in a second cohort cultured to 6
 weeks post-fertilization continued to display hormetic growth responses,
 whereas benefits for heat tolerance were lost; in contrast, negative
 effects of short-term exposure to extreme heat stress (39°C) persisted.
 These results demonstrate that some dose-dependent effects of priming
 waned over time while others persisted, resulting in heterogeneity in
 organismal performance across ontogeny following priming. Overall, these
 findings suggest that heat priming may augment the climate resilience of
 marine invertebrate early life stages via the modulation of key
 developmental and physiological phenotypes, while also affirming the need
 to limit further anthropogenic ocean warming.
description: All details are present in the manuscript and code comments.
description: # Data and code for: Acute heat priming promotes short-term climate
 resilience of early life stages in a model sea anemone (Glass et al.)
 Dataset includes files for: 1. Data pertaining to the development of
 Nematostella vectensis larvae past the planula stage 2. Data pertaining to
 dose-response curves (DRC) quantifying survival of larvae after exposure
 to heat ramps 3. Data pertaining to the expression of heat shock protein
 70 (HSP70) in larvae at 11 days post-fertilization (DPF) 4. Data
 pertaining to the long-term body column lengths and tentacles numbers of
 larvae and juveniles through 6 weeks post-fertilization (WPF) 5. Data
 pertaining to the heat tolerance (survival) of juveniles at 6 WPF
 following heat shock 6. Data pertaining to the lethal temperature 50s
 (LT50s) displayed by larvae derived from DRCs after heat shock 7. Data
 pertaining to the respiration rates and protein content of larvae 8. Data
 pertaining to the sizes of larvae from 4-11 DPF ## Data and file structure
 ### Files 1 and 2: Development\_data.xlsx and .csv These files are for
 data pertaining the development of Nematostella vectensis larvae past the
 planula stage These files have headers, which include: * priming
 temperature (°C) = temperature at which larvae were primed for 1 hour at 3
 DPF * group = arbitrary number assigned to the replicate dishes in which
 larvae were cultured * days post-fertilization = self-explanatory *
 percent/planula count = percent/number of larvae in the planula stage *
 percent/post-planula count = percent/number of larvae past the planula
 stage ### Files 3 and 4: DRC\_data.xlsx and .csv These files are for data
 pertaining to dose-response curves (DRC) quantifying survival of larvae
 after exposure to heat ramps These files have headers, which include: *
 priming temperature (°C) = temperature at which larvae were primed for 1
 hour at 3 DPF * group = arbitrary number assigned to the replicate dishes
 in which larvae were cultured * days post-fertilization = self-explanatory
 * treatment temperature (°C) = peak temperature to which larvae were
 exposed for 1 hour during a heat ramp * larvae surviving = number of
 larvae alive 48 hours post-treatment * total larvae = number of larvae
 originally treated * proportion/percent surviving = larvae surviving/total
 larvae ### Files 5 and 6: HSP70\_data.xlsx and .csv These files are for
 data pertaining to the expression of heat shock protein 70 (HSP70) in
 larvae at 11 days post-fertilization (DPF) These files have headers, which
 include: * priming temperature (°C) = temperature at which larvae were
 primed for 1 hour at 3 DPF * group = arbitrary number assigned to the
 replicate dishes in which larvae were cultured * days post-fertilization =
 self-explanatory * HSP70 = mean gray value for Western blot band
 representing HSP70 as determined via ImageJ * tubulin = mean gray value
 for Western blot band representing tubulin as determined via ImageJ *
 normalized HSP70 = HSP70/tubulin * LT50 (°C) = lethal temperature 50 for
 larvae at 11 DPF, copied from LT50 data file ### Files 7 and 8:
 Long\_term\_growth\_data.xlsx and .csv These files are for data pertaining
 to the long-term body column lengths and tentacles numbers of larvae and
 juveniles through 6 weeks post-fertilization (WPF) These files have
 headers, which include: * priming temperature (°C) = temperature at which
 larvae were primed for 1 hour at 3 DPF * group = arbitrary number assigned
 to the replicate dishes in which larvae were cultured * weeks
 post-fertilization = self-explanatory * body column length/width (cm/mm) =
 lengths and widths of larval/juvenile body columns as determined from
 images via ImageJ * aspect ratio = length/width * volume/surface area
 (mm^3/mm^2) = calculated from lengths and widths * surface area to volume
 ratio = surface area/volume * tentacles = number of tentacles possessed by
 juveniles ### Files 9 and 10: Long\_term\_heattol\_data.xlsx and .csv
 These files are for data pertaining to the heat tolerance (survival) of
 juveniles at 6 WPF following heat shock These files have headers, which
 include: * priming temperature (°C) = temperature at which larvae were
 primed for 1 hour at 3 DPF * group = arbitrary number assigned to the
 replicate dishes in which larvae were cultured * juveniles surviving =
 number of juveniles surviving at 21 hours following heat shock * total
 juveniles = number of juveniles originally exposed to heat shock *
 survival (%) = juveniles surviving/total juveniles\*100 ### Files 11 and
 12: LT50\_data.xlsx and .csv These files are for data pertaining to the
 lethal temperature 50s (LT50s) displayed by larvae derived from DRCs after
 heat shock These files have headers, which include: * priming temperature
 (°C) = temperature at which larvae were primed for 1 hour at 3 DPF * group
 = arbitrary number assigned to the replicate dishes in which larvae were
 cultured * days post-fertilization = self-explanatory * LT50 (°C) = lethal
 temperature as determined from dose-response curves of survival following
 acute heat shock ### Files 13 and 14: Respiration\_and\_protein\_data.xlsx
 and .csv These files are for data pertaining to the respiration rates and
 protein content of larvae These files have headers, which include: *
 priming temperature (°C) = temperature at which larvae were primed for 1
 hour at 3 DPF * group = arbitrary number assigned to the replicate dishes
 in which larvae were cultured * days post-fertilization = self-explanatory
 * respiration (nmol O2 per minute) = respiration rate determined by taking
 the slope of raw data from respirometer * number of larvae = number of
 larvae in well of respirometry plate * respiration (nmol O2 per minute per
 larva) = respiration (nmol O2 per minute)/number of larvae * protein (ug)
 = protein content of larvae in well as determined via Bradford assays *
 protein (ug per larva) = protein (ug)/number of larvae * respiration (nmol
 O2 per minute per protein) = respiration (nmol O2 per minute per
 larva)/protein (ug) * respiration (pmol O2 per minute per protein) =
 respiration (nmol O2 per minute per protein)\*1000 ### Files 15 and 16:
 Size\_data.xlsx and .csv These files are for data pertaining to the sizes
 of larvae from 4-11 DPF These files have headers, which include: * priming
 temperature (°C) = temperature at which larvae were primed for 1 hour at 3
 DPF * group = arbitrary number assigned to the replicate dishes in which
 larvae were cultured * weeks post-fertilization = self-explanatory * body
 column length/width (cm/mm) = lengths and widths of larval body columns as
 determined from images via ImageJ * aspect ratio = length/width *
 volume/surface area (mm^3/mm^2) = calculated from lengths and widths *
 surface area to volume ratio = surface area/volume ## Code/Software ###
 Filse 1 and 2: Nematostella HP script for manuscript.R These files are an
 R script (markdown and PDF forms) that was used to analyze and generate
 plots of data, and can be run using the .csv files.