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Effects of dietary fatty acids on mitochondrial phospholipid compositions, oxidative status and mitochondrial gene expression of zebrafish at different ages

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Abstract

Mitochondrial decay is generally associated with impairment in the organelle bioenergetics function and increased oxidative stress, and it appears that deterioration of mitochondrial inner membrane phospholipids (PL) and accumulation of mitochondrial DNA (mtDNA) mutations are among the main mechanisms involved in this process. In the present study, mitochondrial membrane PL compositions, oxidative status (TBARS content and SOD activity) and mtDNA gene expression of muscle and liver were analyzed in zebrafish fed two diets with lipid supplied either by rapeseed oil (RO) or a blend 60:40 of RO and DHA500 TG oil (DHA). Two feeding trials were performed using zebrafish from the same population of two ages (8 and 21 months). Dietary FA composition affected fish growth in 8-month-old animals, which could be related to an increase in stress promoted by diet composition. Lipid peroxidation was considerably higher in mitochondria of 8-month-old zebrafish fed the DHA diet than in animals fed the RO diet. This could indicate higher oxidative damage to mitochondrial lipids, very likely due to increased incorporation of DHA in PL of mitochondrial membranes. Lipids would be among the first molecules affected by mitochondrial reactive oxygen species, and lipid peroxidation could propagate oxidative reactions that would damage other molecules, including mtDNA. Mitochondrial lipid peroxidation and gene expression of 21-month-old fish showed lower responsiveness to diet composition than those of younger fish. Differences found in the effect of diet composition on mitochondrial lipids between the two age groups could be indicating age-related changes in the ability to maintain structural homeostasis of mitochondrial membranes.

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Abbreviations

B2M:

β-2-Microglobulin

BACT:

β-Actin

BHT:

Butylated hydroxytoluene

cDNA:

Complementary DNA

CL:

Cardiolipin

COX:

Cytochrome c oxidase complex

DHA:

Docosahexaenoic acid

ETC:

Electron transport chain

FA:

Fatty acid

FAME:

Fatty acid methyl esters

HP-TLC:

High-performance thin-layer chromatography

LA:

Linoleic acid

LC-PUFA:

Long-chain polyunsaturated fatty acid

MIM:

Mitochondrial inner membrane

mtDNA:

Mitochondrial DNA

MUFA:

Monounsaturated fatty acids

NAC:

No-amplification control

ND:

NADH-coenzyme Q oxidoreductase complex

NTC:

No-template control

OA:

Oleic acid

PC:

Phosphatidylcholine

PE:

Phosphatidylethanolamine

PI:

Phosphatidylinositol

PIn:

Peroxidation index

PL:

Phospholipid

PS:

Phosphatidylserine

PUFA:

Polyunsaturated fatty acid

qPCR:

Quantitative PCR

RO:

Rapeseed oil

ROS:

Reactive oxygen species

SFA:

Saturated fatty acids

SM:

Sphingomyelin

RT-PCR:

Real-time PCR

SEM:

Standard error of the mean

SGR:

Specific growth rate

SOD:

Superoxide dismutase

TBARS:

Thiobarbituric acid reactive substances

TBA:

Thiobarbituric acid

TCA:

Trichloroacetic acid

TLC:

Thin-layer chromatography

Tm:

Melting temperature

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Acknowledgments

This research and P.F.A.-P. were funded by a Marie Curie Intra-European Fellowship within the 7th Community Framework Programme (PIEF-GA-2011-297964, OLDMITO). The authors report no conflicts of interest.

Ethical standard

The authors confirm that there is no potential conflict of interests. Fish were treated in accordance with British national ethical requirements and the experiments conducted under the UK Government Home Office project Licence number PPL 60/03969 in accordance with the amended Animals Scientific Procedures Act 1986 implementing EU directive 2010/63.

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    1. School of Natural Sciences, Institute of Aquaculture, University of Stirling, Stirling, FK9 4LA, Scotland, UK

      M. B. Betancor, P. F. Almaida-Pagán, A. Hernández & D. R. Tocher

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    1. M. B. Betancor
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    2. P. F. Almaida-Pagán
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    3. A. Hernández
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    4. D. R. Tocher
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    Correspondence to M. B. Betancor.

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    M. B. Betancor and P. F. Almaida-Pagán have contributed equally to this work.

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    Betancor, M.B., Almaida-Pagán, P.F., Hernández, A. et al. Effects of dietary fatty acids on mitochondrial phospholipid compositions, oxidative status and mitochondrial gene expression of zebrafish at different ages. Fish Physiol Biochem 41, 1187–1204 (2015). https://doi.org/10.1007/s10695-015-0079-0

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