Submitted on March 29, 2005
Revised on April 22, 2005
Accepted on April 23, 2005
In situ perfusion protocol of rat epididymal adipose tissue useful for metabolic studies
Gemma Cònsol-Urgellés, Anna Moles, David Ricart-Jané, and Miquel Llobera
Bioquímica i Biologia Molecular, Universitat de Barcelona, Barcelona, Catalunya 08071
Corresponding Author: 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, whilst 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 due to 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 and we present it here as a protocol in order 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 perfusion of epididymal white adipose tissue for use in metabolic studies.
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