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-hydroxylase: a potential
O2 sensor in rat arterioles and skeletal muscle
cells
Medical College of Wisconsin and Marquette University, Milwaukee, Wisconsin 53226
The purposes
of this study were to 1) further evaluate the possible role
that vasoconstrictor metabolites of cytochrome P-450 (CYP)
-hydroxylase plays in O2-induced constriction of
arterioles in the rat skeletal muscle microcirculation, 2)
determine whether -hydroxylases are expressed in rat cremaster
muscle, and 3) determine whether the enzyme is located in
the parenchyma or the arterioles. O2-induced constriction
of third-order arterioles in the in situ cremaster muscle of
Sprague-Dawley rats was significantly inhibited by the CYP inhibitors
N-methyl-sulfonyl-12,12-dibromododec-11-enamide (DDMS; 50 µM) and 17-octadecynoic acid (ODYA; 10 µM). Immunoblot analysis
with antibody raised against CYP4A protein indicated the presence of
immunoreactive proteins in the cremaster muscle and in isolated
arterioles and muscle fibers from this tissue. However, the molecular
mass of the immunoreactive proteins was 85 kDa instead of the expected
50-52 kDa for CYP4A -hydroxylase isolated from rat liver or
kidney. Treatment of the cremaster muscle with deglycosidases
shifted the bands to the expected range which indicates that these
proteins are likely glycosylated in skeletal muscle.
Immunohistochemistry revealed intense staining of both muscle fibers
and microvessels in the cremaster muscle. The results of this study
indicate that O2 sensing in the skeletal muscle
microcirculation may be mediated by CYP4A -hydroxylases in both
arterioles and parenchymal cells.
microcirculation; 20-hydroxyeicosatetraenoic acid; autoregulation; vascular smooth muscle
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