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Vol. 290, Issue 1, 452-463, July 1999
1-Adrenoceptors on Vascular
Smooth Muscle: Correlation with the Regulation of
Contraction1
The Department of Pharmacology (S.L.H., S.E.E., D.F.M., J.R.O.,
R.W.H., M.T.P.) and the Vascular Biology Research Group (S.L.H.,
S.E.E., D.F.M., J.R.O., M.T.P.), The University of Kentucky College of
Medicine, Lexington, Kentucky; The Department of Molecular Cardiology,
Lerner Research Institute, The Cleveland Clinic Foundation, Cleveland,
Ohio (D.M.P.)
Previous work has shown that the genes encoding each
1-adrenoceptor subtype are coexpressed throughout the
peripheral vascular system. We have evaluated subtype-selective
antibodies as tools to determine the extent of protein expression in
arteries. The 1A-, 1B-, and
1D-adrenoceptors were detected in the medial layer of
the aorta, caudal, femoral, iliac, renal, superior mesenteric, and
mesenteric resistance arteries. In Rat1 fibroblasts expressing each
subtype, immunoreactivity was noted both on the cell surface and in a
perinuclear orientation. Intense 1B-adrenoceptor
immunostaining was similarly localized in cultured femoral and renal
vascular smooth muscle cells. Although the cellular localization
appeared to be the same, immunoreactivity obtained with
1A- and 1D-adrenoceptors was much less
intense than that with the 1B-adrenoceptor. The 1A-adrenoceptor selective agonist A-61603 was 22-fold
more potent in activating renal artery contraction when compared with
the femoral artery. The expression of each
1-adrenoceptor was significantly decreased by in vivo
application of antisense oligonucleotides targeted against each
subtype. Inhibition of the expression of only one, the
1A in renal and the 1D in femoral
arteries, reduced the contractile response to naphazoline. The results
show: 1) subtype-selective antibodies can be used in tissues and cell
culture to localize the 1-adrenoceptor subtypes, 2) in
addition to expression on the cell surface, the
1-adrenoceptors are expressed intracellularly, and 3)
despite expression of all adrenoceptors, a single subtype mediates the
contractile response in the femoral and renal arteries.
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