Thakali Keshari, Galligan James J, Fink Gregory D, Gariepy Cheryl E, Watts Stephanie W
Department of Pharmacology and Toxicology, Michigan State University, East Lansing, MI 48823, USA.
Vascul Pharmacol. 2008 Jul;49(1):6-13. doi: 10.1016/j.vph.2008.03.005. Epub 2008 Apr 3.
Heterodimerization of G-protein coupled receptors can alter receptor pharmacology. ET A and ET B receptors heterodimerize when co-expressed in heterologous expression lines. We hypothesized that ET A and ET B receptors heterodimerize and pharmacologically interact in vena cava from wild-type (WT) but not ET B receptor deficient (sl/sl) rats. Pharmacological endothelin receptor interaction was assessed by comparing ET-1-induced contraction in rings of rat thoracic aorta and thoracic vena cava from male Sprague Dawley rats under control conditions, ET A receptor blockade (atrasentan, 10 nM), ET B receptor blockade (BQ-788, 100 nM) or ET B receptor desensitization (Sarafotoxin 6c, 100 nM) and ET A plus ET B receptor blockade or ET A receptor blockade plus ET B receptor desensitization. In addition, similar pharmacological ET receptor antagonism experiments were performed in rat thoracic aorta and vena cava from WT and sl/sl rats. ET A but not ET B receptor blockade or ET B receptor desensitization inhibited aortic and venous ET-1-induced contraction. In vena cava but not aorta, when ET B receptors were blocked (BQ-788, 100 nM) or desensitized (S6c, 100 nM), atrasentan caused a greater inhibition of ET-1-induced contraction. Vena cava from WT but not sl/sl rats exhibited similar pharmacological ET receptor interaction. Immunocytochemistry was performed on freshly dissociated aortic and venous vascular smooth muscle cells to determine localization of ET A and ET B receptors. ET A and ET B receptors qualitatively co-localized more strongly to the plasma membrane of aortic compared to venous vascular smooth muscle cells. Our data suggest that pharmacological ET A and ET B receptor interaction may be dependent on the presence of functional ET B receptors and independent of receptor location.
G蛋白偶联受体的异源二聚化可改变受体药理学特性。当在异源表达系统中共表达时,ET A和ET B受体会形成异源二聚体。我们推测,在野生型(WT)大鼠而非ET B受体缺陷型(sl/sl)大鼠的腔静脉中,ET A和ET B受体会形成异源二聚体并在药理学上相互作用。通过比较在对照条件下、ET A受体阻断(阿曲生坦,10 nM)、ET B受体阻断(BQ-788,100 nM)或ET B受体脱敏(铃蟾毒素6c,100 nM)以及ET A加ET B受体阻断或ET A受体阻断加ET B受体脱敏情况下,雄性Sprague Dawley大鼠胸主动脉和胸段腔静脉环中ET-1诱导的收缩,来评估药理学上的内皮素受体相互作用。此外,在WT和sl/sl大鼠的胸主动脉和腔静脉中进行了类似的药理学ET受体拮抗实验。ET A受体阻断而非ET B受体阻断或ET B受体脱敏可抑制主动脉和静脉中ET-1诱导的收缩。在腔静脉而非主动脉中,当ET B受体被阻断(BQ-788,100 nM)或脱敏(S6c,100 nM)时,阿曲生坦对ET-1诱导的收缩具有更大的抑制作用。WT大鼠而非sl/sl大鼠的腔静脉表现出类似的药理学ET受体相互作用。对新鲜分离的主动脉和静脉血管平滑肌细胞进行免疫细胞化学检测,以确定ET A和ET B受体的定位。与静脉血管平滑肌细胞相比,ET A和ET B受体在主动脉细胞膜上的定性共定位更强。我们的数据表明,药理学上的ET A和ET B受体相互作用可能依赖于功能性ET B受体的存在,而与受体位置无关。