Expression and characterization of the human alpha 2B-adrenoceptor in a vascular smooth muscle cell line.
A vascular smooth muscle cell line stably expressing the human alpha 2B-adrenoceptor at a density of 1.5 pmol/mg membrane protein was generated by transfection of rat A7r5 cells. [35S]GTPgammaS binding experiments and [3H]thymidine incorporation experiments indicated that the expressed receptors were functional, had the expected pharmacological characteristics and efficiently stimulated smooth muscle cell proliferation. Confocal fluorescence microscopy was used to visualize alpha2B-adrenoceptors in A7r5-alpha 2B cells and indicated that the receptors were mainly localized in the plasma membrane. The expression of the smooth muscle-specific marker alpha-actin was similar in transfected A7r5-alpha 2B cells and in non-transfected A7r5 wild-type cells. The generated A7r5-alpha 2B cell line will be a useful tool for studying the function and regulation of alpha 2B-adrenoceptors in vascular smooth muscle cells.
Huhtinen
Anna
A
Department of Pharmacology, Drug Development and Therapeutics, University of Turku, FI-20520 Turku, Finland. anna.huhtinen@utu.fi
Scheinin
Mika
M
D001711:Biotransformation / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003627:Data Interpretation, Statistical; D005455:Fluorescent Antibody Technique; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008856:Microscopy, Fluorescence; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D011485:Protein Binding; D043704:Receptor, Adenosine A2B / Q000096:biosynthesis* / Q000235:genetics; D013936:Thymidine / Q000378:metabolism
D008024:Ligands; D043704:Receptor, Adenosine A2B; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D013936:Thymidine