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Functional characterisation of UCP1 in the common carp: uncoupling activity in liver mitochondria and cold-induced expression in the brain

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Abstract

Mammalian uncoupling protein 1 (UCP1) mediates nonshivering thermogenesis in brown adipose tissue. We previously reported on the presence of a UCP1 orthologue in ectothermic fish and observed downregulation of UCP1 gene expression in the liver of the common carp. Neither the function of UCP1, nor the mode of UCP1 activation is known in carp liver mitochondria. Here, we compared the proton conductance at 25°C of liver mitochondria isolated from carp either maintained at 20°C (warm-acclimated, WA) or exposed to 8°C (cold-acclimated, CA) water temperature for 7–10 days. Liver mitochondria from WA carp had higher state four rates of oxygen consumption and greater proton conductance at high membrane potential. Liver mitochondria from WA, but not from CA, carp showed a strong increase in proton conductance when palmitate (or 4-hydroxy-trans-2-nonenal, HNE) was added, and this inducible proton conductance was prevented by addition of GDP. This fatty acid sensitive proton leak is likely due to the expression of UCP1 in the liver of WA carp. The observed biochemical properties of proton leak strongly suggest that carp UCP1 is a functional uncoupling protein with broadly the same activatory and inhibitory characteristics as mammalian UCP1. Significant UCP1 expression was also detected in our previous study in whole brain of the carp. We here observed a twofold increase of UCP1 mRNA in carp brain following cold exposure, suggesting a role of UCP1 in the thermal adaptation of brain metabolism. In situ hybridization located the UCP1 gene expression to the optic tectum responsible for visual system control, the descending trigeminal tract and the solitary tract. Taken together, this study characterises uncoupling protein activity in an ectotherm for the first time.

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Abbreviations

ADP:

Adenosine 5′-(trihydrogen diphosphate)

BSA:

Bovine serum albumin

CA:

Cold-acclimated

DEPC:

Diethylpyrocarbonate

DTT:

Dithiothreitol

EDTA:

Ethylenediaminetetraacetic acid

EGTA:

Ethylene glycol bis (2-aminoethyl ether)-N,N,N′N′-tetraacetic acid

FCCP:

Carbonyl cyanide p-trifluoro-methoxyphenylhydrazone

GDP:

Guanosine 5′-diphosphate

HEPES:

4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid

PBS:

Phosphate buffered saline

SSC:

Sodium chloride sodium citrate buffer

35S-UTP:

Uridine 5′-[α-35S]thiotriphosphate

TEA:

Triethanolamine

TPMP:

Triphenylmethylphosphonium

Tris–HCl:

(Hydroxymethyl) aminomethane-hydrochloride

WA:

Warm-acclimated

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Acknowledgments

We thank Helen Boysen, Ian Goldstone and Sigrid Stöhr for excellent technical assistance, and the Department of Zoology, Cambridge University for providing us with their animal facilities. This study was funded by the Medical Research Council, the DAAD and the DFG (Grant KL 973/7).

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Authors and Affiliations

  1. Animal Physiology, Department of Biology, Philipps University Marburg, Karl-von-Frisch-Str. 8, Marburg, 35032, Germany

    Martin Jastroch, Michael Helwig & Martin Klingenspor

  2. Medical Research Council Dunn Human Nutrition Unit, Hills Road, Cambridge, CB2 2XY, UK

    Julie A. Buckingham & Martin D. Brand

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  1. Martin Jastroch
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  2. Julie A. Buckingham
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  3. Michael Helwig
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  4. Martin Klingenspor
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  5. Martin D. Brand
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Correspondence to Martin Jastroch.

Additional information

Communicated by G. Heldmaier.

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360_2007_171_MOESM1_ESM.pdf

Suppl. Photograph Photograph of carp either acclimated to 20°C or 8°C. The body cavity was opened to illustrate altered morphology of the liver due to cold exposure (PDF 85 kb)

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Jastroch, M., Buckingham, J.A., Helwig, M. et al. Functional characterisation of UCP1 in the common carp: uncoupling activity in liver mitochondria and cold-induced expression in the brain. J Comp Physiol B 177, 743–752 (2007). https://doi.org/10.1007/s00360-007-0171-6

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