An Innovative Setup for High-Throughput Respirometry of Small Aquatic Animals

Drown, Melissa K.; DeLiberto, Amanda N.; Crawford, Douglas L.; Oleksiak, Marjorie F.

doi:10.1101/2020.01.20.912469

Cited by 6 publications

(6 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…All data are available at the Dryad Digital Repository ( ) [ 102 ]. All scripts used in analysis can be found on github: and have been archived with the Zenodo repository: .…”

Section: Data Accessibilitymentioning

confidence: 99%

Interindividual plasticity in metabolic and thermal tolerance traits from populations subjected to recent anthropogenic heating

et al. 2021

Self Cite

View full text Add to dashboard Cite

To better understand temperature's role in the interaction between local evolutionary adaptation and physiological plasticity, we investigated acclimation effects on metabolic performance and thermal tolerance among natural Fundulus heteroclitus (small estuarine fish) populations from different thermal environments. Fundulus heteroclitus populations experience large daily and seasonal temperature variations, as well as local mean temperature differences across their large geographical cline. In this study, we use three populations: one locally heated (32°C) by thermal effluence (TE) from the Oyster Creek Nuclear Generating Station, NJ, and two nearby reference populations that do not experience local heating (28°C). After acclimation to 12 or 28°C, we quantified whole-animal metabolic (WAM) rate, critical thermal maximum (CT max ) and substrate-specific cardiac metabolic rate (CaM, substrates: glucose, fatty acids, lactate plus ketones plus ethanol, and endogenous (i.e. no added substrates)) in approximately 160 individuals from these three populations. Populations showed few significant differences due to large interindividual variation within populations. In general, for WAM and CT max , the interindividual variation in acclimation response (log 2 ratio 28/12°C) was a function of performance at 12°C and order of acclimation (12–28°C versus 28–12°C). CT max and WAM were greater at 28°C than 12°C, although WAM had a small change (2.32-fold) compared with the expectation for a 16°C increase in temperature (expect 3- to 4.4-fold). By contrast, for CaM, the rates when acclimatized and assayed at 12 or 28°C were nearly identical. The small differences in CaM between 12 and 28°C temperature were partially explained by cardiac remodeling where individuals acclimatized to 12°C had larger hearts than individuals acclimatized to 28°C. Correlation among physiological traits was dependent on acclimation temperature. For example, WAM was negatively correlated with CT max at 12°C but positively correlated at 28°C. Additionally, glucose substrate supported higher CaM than fatty acid, and fatty acid supported higher CaM than lactate, ketones and alcohol (LKA) or endogenous. However, these responses were highly variable with some individuals using much more FA than glucose. These findings suggest interindividual variation in physiological responses to temperature acclimation and indicate that additional research investigating interindividual may be relevant for global climate change responses in many species.

show abstract

“…All data are available at the Dryad Digital Repository ( ) [ 102 ]. All scripts used in analysis can be found on github: and have been archived with the Zenodo repository: .…”

Section: Data Accessibilitymentioning

confidence: 99%

Interindividual plasticity in metabolic and thermal tolerance traits from populations subjected to recent anthropogenic heating

et al. 2021

Self Cite

View full text Add to dashboard Cite

show abstract

“…We additionally thank the reviewers and editor for their thoughtful comments that improved the manuscript. This manuscript has been released as a pre-print at BixRxiv (Drown et al, 2020).…”

Section: Discussionmentioning

confidence: 99%

An Innovative Setup for High-Throughput Respirometry of Small Aquatic Animals

et al. 2020

View full text Add to dashboard Cite

Metabolic rate is often measured as a phenotype in evolutionary genetics, among other fields including many facets of physiology, behavior, and ecology, because it impacts organismal fitness, is repeatable and heritable, and is responsive to numerous environmental variables. Aquatic respirometry, a method used to measure metabolic rate, has allowed key questions in these fields to be investigated, namely: (1) why do individuals from the same population exhibit up to 3-fold differences in metabolic rate, (2) how does metabolic rate change during an individual's lifetime, and ( 3) what metabolic rate is advantageous in a specific environment? Current respirometry studies often suffer from small sample sizes and rely on low throughput approaches to measure metabolic rate, making it difficult to answer these and other relevant ecological and evolutionary questions due to lack of power, failure to capture true biological variation, and confounding variables, like time, that are introduced due to limitations in methodology.Here we describe a scalable high-throughput intermittent flow respirometer (HIFR) design and use it to measure the metabolic rates of 19 aquatic animals in one night while reducing equipment costs and time by more than 50%.

show abstract

“…Individuals acclimated to 12°C and 28°C for at least 4 weeks were measured for whole animal metabolic rate (overnight intermittent flow respirometry) with a minimum of 20 replicate metabolic rate measures per individual used to determine the standard metabolic rate (SMR) in mgO2 hr -1 (7). Critical thermal maximum (CT max ) was measured in a 10-gallon tank that was slowly heated at a rate of 0.3°C min -1 as in (86) and was defined as the point when fish lost equilibrium in the water column for 5 consecutive seconds.…”

Section: Methodsmentioning

confidence: 99%

“…In comparison to inbred lines, non-model organisms have more polymorphic alleles and greater heterozygosity, and thus non-model organism studies can provide a more nuanced understanding of how natural genetic variation impacts complex phenotypes (5,6). Furthermore, non-model organism studies have become even more relevant with increased genomic resource availability and development of high-throughput approaches to measure behavioral and physiological traits under realistic ecological conditions (7)(8)(9)(10).…”

Section: Introductionmentioning

confidence: 99%

mRNA expression explains metabolic and thermal physiology

Drown

Crawford

Oleksiak

2022

Preprint

Self Cite

View full text Add to dashboard Cite

Quantifying mRNA expression, which is heritable and physiologically inducible, reveals biologically important networks and pathways underlying complex traits. Here, we quantify mRNA expression in Fundulus heteroclitus, a small teleost fish, among three populations acclimated to 12°C and 28°C and relate it to variation in six, complex, physiological traits (whole animal metabolism (WAM), critical thermal maximum (CT max), and four substrate specific cardiac metabolic rates (CaM)). Although 366 heart mRNAs and 528 brain mRNAs had significant differential expression between the two acclimation temperatures, none of the mRNA acclimation responses were shared across all three populations in any tissue. Yet, within an acclimation temperature across all three populations, weighted gene co-expression network analyses show that mRNA expression patterns explained WAM, CT max, and CaM trait variation. These analyses revealed 9 significant heart MEs (first principal component of module expression) and 4 significant brain MEs. Heart MEs explained variation in WAM, CT max, and two of the four substrate specific cardiac metabolic rates at 12°C, and CT max at 28C. In contrast, brain MEs explained CT max and WAM at 28°C but not at 12°C. Combining MEs as multiple correlations, 82% of variation in WAM at 12°C was explained by four heart MEs, 80% of variation in fatty-acid CaM at 12°C was explained by three heart MEs, and 72% of variation in CT max at 28°C was explained by three brain MEs. These MEs were enriched for Kyoto Encyclopedia of Genes and Genomes (KEGG) terms related to specific metabolic pathways, suggesting that they represent biologically relevant pathways. Together these data suggest that mRNA co-expression explains complex traits; moreover, physiological traits are more reliant on heart expression at 12°C and brain expression at 28°C.

show abstract

An Innovative Setup for High-Throughput Respirometry of Small Aquatic Animals

Cited by 6 publications

References 31 publications

Interindividual plasticity in metabolic and thermal tolerance traits from populations subjected to recent anthropogenic heating

Interindividual plasticity in metabolic and thermal tolerance traits from populations subjected to recent anthropogenic heating

An Innovative Setup for High-Throughput Respirometry of Small Aquatic Animals

mRNA expression explains metabolic and thermal physiology

Contact Info

Product

Resources

About