DOI: 10.5061/dryad.41ns1rnt6
title: Context-dependent relationships between metabolism and behavior across temperature acclimations in sheepshead minnows (<em>Cyprinodon variegatus</em>)

description: Marine fish are able to cope changes in their environment through altering
 their physiology and behaviour. Among environmental stressors, thermal
 stress is particularly relevant for fish because increased temperature
 leads to elevated baseline metabolic costs and also has known impacts on
 behavior. Whether behavioral and physiological responses to warming are
 linked has become a major area of interest within the broader study of
 animal behavior. The Performance Model provides a conceptual framework
 linking metabolism and behavior such that individual differences in
 baseline metabolic demand influence behavioral expression specifically
 related to acquiring resources that support metabolic machinery. Thus,
 under warming, increased energy demand should lead to increased
 risk-taking and lower behaviours that deter predation (e.g., shoaling and
 scototaxis). Alternatively, behavior related to risk-taking has been
 linked to differences in physiological stress, particularly cortisol. On
 this background, our objective was to assess behavioral and metabolic
 responses to warming and the relationship between behaviour and metabolic
 rate across two temperatures in an estuarine model teleost, the sheepshead
 minnow (Cyprinodon variegatus). Fish were acclimated to 22°C or 32°C for
 14-days and standard metabolic rate (SMR) was measured. A separate group
 of fish were acutely raised to 32°C following acclimation at the control
 temperature. Each fish subsequently tested in a series of behavioural
 assays to assess activity, exploration, sociability, and anxiety-like
 behavior. As anticipated, warmer temperatures increased in SMR and
 increased activity while reducing anxiety-like behavior. However,
 relationships between behavior and metabolism were present only in control
 conditions and dissipated under the warming. We sought to explain
 differences in behavioral-metabolic patterns across the two temperatures
 through mitochondrial performance and baseline cortisol. We argue that the
 behavioral-metabolic relationships are sensitive to context and
 environmental factors need to be considered when exploring the
 intersection between behavior and physiology.
description: # Context-dependent relationships between metabolism and behavior across
 temperature acclimations in sheepshead minnows (*Cyprinodon variegatus*)
 Dataset DOI: [10.5061/dryad.41ns1rnt6](10.5061/dryad.41ns1rnt6) ##
 Description of the data and file structure These data were used to assess
 relationships between behavior and metabolism among fish acclimated to 22
 degrees C or 32 degrees C. We additionally aimed to explore these
 relationships with an assessment of brain mitochondrial respirometry and
 baseline cortisol analysis. Multiple datasets are included for behavior
 and whole-body respiration, mitochondrial respiration, and baseline
 cortisol. ### Files and variables #### File:
 2023_minnows_TempAccl_RespBehaviorPC_Aug2025.csv **Description:** Behavior
 and metabolic data across two temperature acclimations (22 and 32 degrees
 Celsius). Blank wells indicate missing data. Note that individuals in the
 acute treatment (acutely transferred from the control temperature to the
 warm temperature during respirometry) were not tested for behavior. #####
 Variables * fishID: individuals * treatment: warm-acclimated, control
 temperature acclimated, acutely transferred from control temperature to
 the warm temperature * sex: male or female * resp_mass: mass in grams of
 the fish at the time of respirometry * mass_start: mass in grams of the
 fish at the beginning of the acclimation window * mass_end: mass in grams
 of the fish following the last experimental assay * TL_end: total length
 (cm) following the final assay * SL_end: standard length (cm) following
 the final assay * SMR: standard metabolic rate * RMR: routine metabolic
 rate * prop_shoal: proportion of total trial time a fish spent within 2-cm
 of a shoal of five mix-sex conspecifics during a sociability assay *
 black_prop: proportion of total trial time that a fish spent in the black
 half of the scototaxis assay * oft_active_percTime: percentage of time
 spent active in the open field test * prop_oft_thig: percentage of time
 spent within 2-cm of the tank walls during the open field test *
 prop_plant: percentage of time spent with a plastic aquarium plant during
 the sociability assay * PC1: composite behavior scores for individuals
 following principal component analysis for the first principal component *
 PC2: composite behavior scores following principal component analysis for
 the second principal component * scaled SMR: standard metabolic rate
 scaled using a scaling exponent of 0.89 #### File:
 2023_MinnowsTempAccl_Mitochondria.csv **Description:** Mitochondrial data
 for fish acclimated to control (22 degrees C) and warm (32 degrees C)
 temperatures. ##### Variables * fishID: individual * treatment: warm or
 control acclimation * sex: male or female * Start (Complex I - LEAK
 State): Oxygen flux (pmol/s/mg) accounting for proton leak in the presence
 of reducing substrates but not ADP. * Complex I + ADP (PHOS State):
 Complex I induced oxygen flux (pmol/s/mg) measured with the addition of
 ADP. * Cytochrome C (Membrane Integrity) * Succinate (CI+CII / PHOS
 State): Oxygen flux (pmol/s/mg) measured after the activation of Complex
 II with the addition of succinate. * OXPHOS: The maximum oxygen
 consumption (pmol/s/g) linked with the oxidation of substrates that
 transfer electrons to oxygen. This process chemiosmotically drives the
 production of ATP. * LEAK: Oxygen consumption (pmol/s/mg) following
 oligomycin-induced inhibition of ATP synthase. Oxygen consumption at this
 state is not coupled with the production of ATP. * Rotenone (Inhibit
 CI): Oxygen consumption (pmol/s/mg) after the inhibition of Complex I with
 the addition of rotenone). * Antimycin A (Inhibit CIII / ROX): Background
 cellular oxygen consumption (pmol/s/mg) measured after the inhibition of
 Complex III with antimycin A. * Coupling (LEAK/OXPHOS): An efficiency
 measure calculated as the ratio of oligomycin-induced leak to OXPHOS. *
 Complex I Coupled (CI/OXPHOS): The contribution of Complex I to OXPHOS
 taken as the ratio of Complex 1 to OXPHOS. * OXPHOS Capacity
 (OXPHOS-LEAK): The difference between OXPHOS and LEAK measures (pmol/s/mg)
 indicating the mitochondrial capacity for oxygen consumption--coupled with
 ATP production--above leak respiration. * RCR: An efficiency measure taken
 as the ratio of OXPHOS to oligomycin-induced LEAK respiration. * OXPHOS
 CE: The OXPHOS control efficiency is calculated as (OXPHOS - LEAK)/OXPHOS.
 #### File: Cortisol_Data.csv **Description:** Fish were acclimated for 2
 weeks at control (22 degrees Celsius) or warm (32 degrees Celsius)
 conditions. Baseline cortisol measures were taken from plasma and assessed
 with an ELISA assay. ##### Variables * treatment: Indicates control or
 warm-acclimated treatments. * cortisol: The baseline cortisol in ng/mL
 **Behavioral videos** **File: oft.zip** **Description:** Open field tests
 for control and warm-acclimated fish. Video files are labelled with the
 fish identification number and assay.  **File: scot.zip** **Description:**
 Scototaxis videos for control and warm-acclimated fish. Video files are
 labelled with the fish identification number and the assay name.  **File:
 soc_control.zip** **Description:** Sociability assay videos for control
 fish. This assay is run in two 10-minute halves, in between which the
 conspecific shoal is transferred to the opposite side of the tank. Video
 files are labelled with the fish identification number and the assay name,
 along with "a" or b" to indicate trial half.  **File:
 soc_warm_acclimated.zip** **Description:** Sociability assay videos for
 warm-acclimated fish. Each file is named with the fish ID, assay type, and
 the half of the assay (a or b). ## Code/software All data were analyzed in
 RStudio (version 2023.03.0+386) with lme4, betareg, FSA, stats, and car
 packages.