Cytochrome P-450 omega-hydroxylase: a potential O(2) sensor in rat arterioles and skeletal muscle cells.
The purposes of this study were to 1) further evaluate the possible role that vasoconstrictor metabolites of cytochrome P-450 (CYP) omega-hydroxylase plays in O(2)-induced constriction of arterioles in the rat skeletal muscle microcirculation, 2) determine whether omega-hydroxylases are expressed in rat cremaster muscle, and 3) determine whether the enzyme is located in the parenchyma or the arterioles. O(2)-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 microM) and 17-octadecynoic acid (ODYA; 10 microM). 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 omega-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 O(2) sensing in the skeletal muscle microcirculation may be mediated by CYP4A omega-hydroxylases in both arterioles and parenchymal cells.
Kunert
M P
MP
Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA.
Roman
R J
RJ
Alonso-Galicia
M
M
Falck
J R
JR
Lombard
J H
JH
D000577:Amides / Q000494:pharmacology; D000818:Animals; D001160:Arterioles / Q000187:drug effects / Q000201:enzymology* / Q000502:physiology; D042926:Cytochrome P-450 CYP4A; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology; D006893:Hydroxyeicosatetraenoic Acids / Q000378:metabolism; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D008297:Male; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000098:blood supply / Q000201:enzymology* / Q000502:physiology; D010100:Oxygen / Q000378:metabolism* / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013450:Sulfones / Q000494:pharmacology; D014661:Vasoconstriction / Q000187:drug effects*
D000577:Amides; D005231:Fatty Acids, Unsaturated; D006893:Hydroxyeicosatetraenoic Acids; D013450:Sulfones; C047008:17-octadecynoic acid; C055987:20-hydroxy-5,8,11,14-eicosatetraenoic acid; D003577:Cytochrome P-450 Enzyme System; C117975:DDMS; D006899:Mixed Function Oxygenases; D042926:Cytochrome P-450 CYP4A; C409986:cytochrome P-450 omega-hydroxylase; D010100:Oxygen