High‐throughput multispecies quantitative polymerase chain reaction assays to study the effects of acute thermal stress in three species of Acipenser sturgeon

Abstract

Fishes experience thermal stress in their natural environment that can be caused by natural or anthropogenic factors and can directly affect their physiology. In this study, we developed a multispecies OpenArray™ qPCR (quantitative polymerase chain reaction) ‘chip’ to measure the effects of acute thermal stress on the mRNA response of Acipenser sturgeons. The qPCR chips were then tested on three species: Acipenser fulvescens , Acipenser oxyrinchus and Acipenser brevirostrum. The A. fulvescens were from two genetically distinct populations (Burntwood River and Winnipeg River, Manitoba, Canada) that were acclimated to three different temperatures (16, 20 and 24°C) to quantify the effects of genetic background and acclimation temperatures on the transcriptomic response to acute thermal stress. Additionally, juvenile A. oxyrinchus oxyrinchus and A. brevirostrum were acclimated to 10°C and then experienced acute thermal stress. All three species were sampled for gill and liver before and after experiencing acute thermal stress, which was applied using a CT max test. The expression of 52 genes of interest was quantified using the multispecies qPCR chip for all three species, along with four reference genes. We showed that acute thermal stress affected the mRNA expression of 17–26 of the thermal metabolic stress genes that were assayed using the chip, depending on the species. However, the mRNA patterns varied between species and genetically distinct populations, suggesting differences in cellular responses to thermal stress that can lead to different susceptibilities to thermal stress in their natural habitats.

Bibliographic Information