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1 Department of Physiology, Medical College of Wisconsin, Milwaukee, Wisconsin 53226; and 2 Department of Biological Sciences and Institute for Biochemistry and Biotechnology, Oakland University, Rochester, Michigan 48309
Only functional studies have suggested the
presence of the ANG II type 2 (AT2) receptor in the
microcirculation. To determine the distribution of this receptor in the
rat skeletal muscle microcirculation, a polyclonal rabbit anti-rat
antiserum was developed and used for immunohistochemistry and Western
blot analysis. The antiserum was prepared against a highly specific and
antigenic AT2-receptor synthetic
peptide and was validated by competition and sensitivity assays.
Western blot analysis demonstrated a prominent, single band at ~40
kDa in cremaster and soleus muscle. Immunohistochemical analysis
revealed a wide distribution of
AT2 receptors throughout the
skeletal muscle microcirculation in large and small microvessels. Microanatomic studies displayed an endothelial localization of the
AT2 receptor, whereas dual
labeling with smooth muscle 
-actin also showed colocalization of the
AT2 receptor with vascular smooth muscle cells. Other cells associated with the microvessels also stained
positive for AT2 receptors.
Briefly, this study confirms previous functional data and localizes the
AT2 receptor to the microcirculation. These studies demonstrate that the
AT2 receptor is present on a
variety of vascular cell types and that it is situated in a fashion
that would allow it to directly oppose ANG II type 1 receptor actions.
immunohistochemistry; vascular smooth muscle cell; endothelial cell; microvessels; angiotensin II
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