Abstract

SNARE proteins are integral to intracellular vesicular trafficking, which in turn is the process underlying the regulated expression of substrate transporters such as the glucose transporter GLUT4 at the cell surface of insulin target tissues. Impaired insulin stimulated GLUT4 trafficking is associated with reduced cardiac function in many disease states, most notably diabetes. Despite this, our understanding of the expression and regulation of SNARE proteins in cardiac tissue and how these may change in diabetes is limited. Here we characterize the array of SNARE proteins expressed in cardiac tissue, and quantify the levels of expression of VAMP2, SNAP23, and Syntaxin4—key proteins involved in insulin-stimulated GLUT4 translocation. We examined SNARE protein levels in cardiac tissue from two rodent models of insulin resistance, db/db mice and high-fat fed mice, and show alterations in patterns of expression are evident. Such changes may have implications for cardiac function.

Highlights

  • Effective regulation of metabolism is essential in all cell types in order to ensure that ATP generation requirements are met

  • The SNARE proteins associated with GLUT4 storage vesicles (GSVs)-plasma membrane (PM) fusion are VAMP2, Syntaxin 4, and SNAP23 [26, 27, 30]

  • Recombinant SNARE proteins were expressed and purified from bacteria and used as standards in quantitative immunoblotting to compare to the signal obtained from mouse cardiac samples

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Summary

Introduction

Effective regulation of metabolism is essential in all cell types in order to ensure that ATP generation requirements are met. This is important within highly energetic organs such as the heart, where the contractile action of cardiomyocytes must be continually fuelled in order to maintain the pumping of approximately 5 liters of oxygen and nutrient rich blood into and around the systemic circulation every minute. Normal cardiac metabolism is characterized by predominant use of fatty acids as a metabolic substrate, with a relatively lower utilization of glucose [1] This is logical as fat is a more abundant and energy rich fuel source, making it ideal for scenarios where sustained moderate levels of ATP are required. Several cardiac disease states are partly defined (and potentially caused) by deficits in glucose uptake and metabolism

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