Abstract

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.

Highlights

  • 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

  • To assess the physiological state and viability of the perfused adipose tissue, we studied the following: a) tissue ATP levels; b) adiponectin secretion; c) the regulation of lipolysis by epinephrine and insulin; and d) the release of LPL by heparin

  • The perfusion of adipose tissue was described by Robert and Scow (1) and Ho and Meng (6) in the 1960s

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Summary

Introduction

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. 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. Based on the modification by Gubser, Di Francesco, and Bickel (7) of the Ho and Meng technique (6), we developed a detailed in situ perfusion method with the aim of providing an reproducible protocol useful for other researchers. This method is appropriate for use in metabolic studies involving the perfusion of different substances and collection of the perfusate through a vein cannula. To assess the physiological state and viability of the perfused adipose tissue, we studied the following: a) tissue ATP levels; b) adiponectin secretion; c) the regulation of lipolysis by epinephrine and insulin; and d) the release of LPL by heparin

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