Sirtuins may mediate temperature-induced metabolic remodeling in threespine stickleback
Data files
Dec 10, 2025 version files 14.53 KB
Abstract
The molecular pathway governing metabolic remodeling in response to temperature in fishes is unknown. We tested the hypothesis that sirtuins, a family of NAD+ dependent lysine deacetylases, drives an increase in aerobic metabolism in response to cold acclimation in stickleback. Stickleback were acclimated to 5, 12 and 20°C and sirtuin activity measured in liver, oxidative and glycolytic muscles using a Promega SIRT-Glo Assay kit. Protein levels of SIRT1 and SIRT3 were quantified using custom antibodies and capillary western blotting. In a second set of experiments, sirtuin activity was inhibited by feeding with EX-527 or activated by feeding with the NAD+ pre-cursor, nicotinamide mononucleotide (NMN). Standard metabolic rate (SMR) was measured using intermittent flow respirometry. Mitochondrial respiration rates were measured in permeabilized liver tissue using a substrate-uncoupler-inhibitor titration protocol . Sirtuin activity significantly increased in response to cold acclimation in liver and oxidative and glycolytic muscles but SIRT3 protein levels declined and SIRT1 protein was undetectable, suggesting one or more other SIRT isoforms contribute to an increase in sirtuin activity in response to cold acclimation. Inhibiting sirtuin activity with EX-527 decreased SMR and the activity of complex I in liver, providing further support for the role of sirtuins in metabolic remodeling in stickleback.
Dataset DOI: 10.5061/dryad.n8pk0p38d
Description of the data and file structure
Stickleback (Gasterosteus aculeatus) were acclimated to 5, 12, and 20°C for 13 weeks. Sirtuin activity was then measured in liver, oxidative, and glycolytic muscles using a Promega SIRT-Glo assay kit, and protein levels of SIRT1 and SIRT3 were quantified using custom antibodies and capillary western blotting. In a second set of experiments, sticklebacks were acclimated to 20°C for 12 weeks. A subset of fish were then moved into an environmental chamber where the temperature in the room was decreased by 3°C per day over 2 days and then by 4°C on day 3 to reach 10°C. After 14 days at 10ºC, bloodworms were supplemented with either the sirtuin inhibitor EX-527 or the sirtuin activator, NMN, mixed with 7% agar for 14 days. Standard metabolic rate was then measured using intermittent respirometry, and mitochondrial function in permeabilized liver tissue was measured using a substrate-uncoupler-inhibitor titration protocol.
Files and variables
File: Physical_characteristics_for_sirtuin_activity_and_NAD(H)_levels.csv
Description: Measurements of physical characteristics of stickleback used to quantify Sirtuin activity and levels of NAD+ and NADH
Variables
- Fish ID: Unique fish identification number
- Temp (C): temperature animal was acclimated to
- Length (cm): standard length of the fish
- mass (g): body mass of the fish
- Condition Factor: = bm/Ls ^3 X100; bm = body mass (g), and Ls = standard length (cm).
File: Sirtuin_activity_and_NAD(H)_levels.csv
Description: Values for measurements of Sirtuin activity and NAD+ and NADH levels liver, oxidative skeletal muscle taken from the pectoral adductor muscle, and white, glycolytic skeletal muscle.
Variables
- Fish ID: Unique fish identification number
- Tissue: tissue type
- Tissue_mass (mg): mass of the tissue
- Sirtuin activity RxnRate_RL/min/ug: activity of sirtuin measured as reaction rate per min per ug protein
- NAD Total (pmol/mg tissue): total nicotinamide adenine dinucleotide plus reduced nicotinamide adenine dinucleotide in pmol per milligram tissue
- NADH (pmol per mg tissue): reduced nicotinamide adenine dinucleotide
- NAD (pmol per mg tissue): nicotinamide adenine dinucleotide measured in pmol per milligram tissue
- NAD:NADH: ratio of ncotinamide adenine dinucleotide -to- reduced nicotinamide adenine dinucleotide
File: Sirtuin_protein_levels.csv
Description: Measurements of protein levels of Sirtuin 3 short and long isoforms in liver, oxidative skeletal muscle taken from the pectoral adductor muscle, and white, glycolytic skeletal muscle (WM). Sirtuin protein levels were normalized to total protein in each sample. Protein levels were quantified using the Compass software (ProteinSimple).
Variables
- Fish_ID liver: Unique fish identification number for the liver sample
- SIRT3_short_normalized liver: amount of the sirtuin 3 short isoform in liver
- SIRT3_long_normalized liver: amount of the sirtuin 3 long isoform in liver
- Fish_ID pect: Unique fish identification number for the pectoral adductor sample
- SIRT3_short_normalized pect: amount of the sirtuin 3 short isoform in the pectoral adductor
- SIRT3_long_normalized pect: amount of the sirtuin 3 long isoform in pectoral adductor
- Fish_ID WM: Unique fish identification number for the white muscle sample
- SIRT3_short_normalized WM: amount of the sirtuin 3 short isoform in white muscle
- SIRT3_long_normalized WM: amount of the sirtuin 3 long isoform in white muscle
File: SMR_and_mito_respiration.csv
Description: Measurements of standard metabolic rate (SMR) using intermittent flow respirometry and respiration rates of mitochondria in permeabilized liver.
Variables
- Fish ID: Unique fish identification number
- Tank: tank number
- Respirometry Chamber: number of respirometry chamber
- Temperature Group: temperature acclimation group of the individual
- Treatment: 10 degree C control or 10 degree C treated with NMN or 10 degree C treated with EX-527, or 20 degree C
- Weight (g) (anesthetized): mass of fish obtained after it was anesthetized
- Length (cm): standard length of fish
- condition factor: bm/Ls ^3 X100; bm = body mass (g), and Ls = standard length (cm).
- OXP I: rate of oxidative phosphorylation through complex I
- OXP II: rate of oxidative phosphorylation through complex II
- OXP I,II: rate of oxidative phosphorylation through complex I and II
- ETS: rate of uncoupled respiration
- CCO: cytochrome c oxidase activity
- Leak Omy: rate of leak respiration in the presence of oligomycin.
- SMR: standard metabolic rate
- Method for calculating SMR: means of the lowest normal distribution (MLND) or 20% quantile
Missing variable indicated with blank cell or N/A