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Originally published In Press as doi:10.1194/jlr.D500016-JLR200 on May 1, 2005

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Journal of Lipid Research, Vol. 46, 1803-1808, August 2005
Copyright © 2005 by American Society for Biochemistry and Molecular Biology

Methods

An in situ perfusion protocol of rat epididymal adipose tissue useful in metabolic studies

Gemma Cònsol, Anna Moles, David Ricart-Jané and Miquel Llobera1

Departament de Bioquímica i Biologia Molecular, Facultat de Biologia, Universitat de Barcelona, E-08071 Barcelona, Spain

Published, JLR Papers in Press, May 1, 2005. DOI 10.1194/jlr.D500016-JLR200

1 To whom correspondence should be addressed. e-mail: millobera{at}ub.edu

Experimental approaches involving the perfusion of tissues and organs offer the advantage of improved physiological relevance over the use of isolated tissues or cells while at the same time being much more controlled and tissue-specific than studies in vivo. Nevertheless, there have been few metabolic studies performed in perfused white adipose tissue, largely because of the difficulty of the surgical technique involved. Although some methods have been described, they are difficult to use as perfusion protocols and their reproducibility is poor. We have modified a rat perfusion method, based on a modification of the Ho and Meng technique, for use with epididymal white adipose tissue (eWAT), and we present it here as a protocol to be reproduced. We also offer surgical solutions for the most common variants of vessel distributions in rats. Using the protocol described here, the perfused adipose tissue is viable and metabolically active, as indicated by the maintenance of tissue ATP levels and adiponectin secretion and by endogenous lipolysis regulation. Moreover, there is a high level of lipoprotein lipase activity in the endothelium of the tissue, which is heparin-releasable.

Thus, this method is a useful and reproducible tool that allows the perfusion of eWAT for use in metabolic studies.

Abbreviations: eWAT, epididymal white adipose tissue; WAT, white adipose tissue

Supplementary key words lipoprotein lipase • heparin • release • lipolysis • adiponectin


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