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Immunolocalization of ACE2 and AT₂ Receptors in Rabbit Atherosclerotic Plaques

Anthony Zulli, Louise M. Burrell, Robert E. Widdop, M. Jane Black, Brian F. Buxton and David L. Hare

Departments of Cardiology (AZ,DLH), Cardiac Surgery (BFB), and Medicine (AZ,LMB), University of Melbourne, Austin Health, Heidelberg, Australia, and Departments of Pharmacology (REW) and Anatomy & Cell Biology (MJB), Monash University, Australia

Correspondence to: Dr. Anthony Zulli, Division of Cardiovascular Research, Department of Cardiology, Austin Health, Heidelberg 3084, Australia. E-mail: azulli{at}unimelb.edu.au

Evidence suggests that angiotensin type 2 receptor (AT₂R) and angiotensin-converting enzyme 2 (ACE2) play a protective role in atherogenesis. These factors have not been identified in rabbit atherosclerotic plaques. Our goal was to localize ACE2 and AT₂R in rabbit atherosclerotic tissues, and determine which cell types express these factors. New Zealand White rabbits were fed either a control diet or a 0.5% cholesterol diet (n=8/group) for 12 weeks. Paraffin-fixed thoracic aorta were serially sectioned and processed for immunohistochemistry using commercially available antibodies to ACE2, AT₂R, RAM 11 (to identify macrophages), and smooth muscle cell actin (SMC) to identify smooth muscle cells and myofibroblasts. AT₂R immunoreactivity, but not ACE2 immunoreactivity, was clearly present in endothelia overlying normal wall. However, both AT₂R and ACE2 immunoreactivity were clearly present in endothelia overlying neo-intima formation and atherosclerotic plaques. Within plaques, both AT₂R and ACE2 immunoreactivity were observed in macrophages and SMC actin-positive cells. Examination of serial sections showed that the majority of cells were both ACE2- and AT₂R-positive. Macrophages and SMC actin-positive cells produce ACE2 and the AT₂R in atherosclerotic plaques. Determining a role for these factors in the control of atherosclerosis will require additional studies. (J Histochem Cytochem 54:147–150, 2006)

Key Words: atherosclerosis • ACE2 • AT₂ receptors • immunohistochemistry

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