Tao Xiaoxiao, Lin Mike T, Thorington Glyne U, Wilson Sean M, Longo Lawrence D, Hessinger David A
Division of Pharmacology, School of Medicine, Loma Linda University, Loma Linda, California;
Division of Physiology, School of Medicine, Loma Linda University, Loma Linda, California; Department of Physiology and Cell Biology, University of South Alabama, Mobile, Alabama.
Am J Physiol Heart Circ Physiol. 2015 Apr 1;308(7):H707-22. doi: 10.1152/ajpheart.00564.2014. Epub 2015 Jan 16.
Acclimatization to high-altitude, long-term hypoxia (LTH) reportedly alters cerebral artery contraction-relaxation responses associated with changes in K(+) channel activity. We hypothesized that to maintain oxygenation during LTH, basilar arteries (BA) in the ovine adult and near-term fetus would show increased large-conductance Ca(2+) activated potassium (BK) channel activity. We measured BK channel activity, expression, and cell surface distribution by use of patch-clamp electrophysiology, flow cytometry, and confocal microscopy, respectively, in myocytes from normoxic control and LTH adult and near-term fetus BA. Electrophysiological data showed that BK channels in LTH myocytes exhibited 1) lowered Ca(2+) set points, 2) left-shifted activation voltages, and 3) longer dwell times. BK channels in LTH myocytes also appeared to be more dephosphorylated. These differences collectively make LTH BK channels more sensitive to activation. Studies using flow cytometry showed that the LTH fetus exhibited increased BK β1 subunit surface expression. In addition, in both fetal groups confocal microscopy revealed increased BK channel clustering and colocalization to myocyte lipid rafts. We conclude that increased BK channel activity in LTH BA occurred in association with increased channel affinity for Ca(2+) and left-shifted voltage activation. Increased cerebrovascular BK channel activity may be a mechanism by which LTH adult and near-term fetal sheep can acclimatize to long-term high altitude hypoxia. Our findings suggest that increasing BK channel activity in cerebral myocytes may be a therapeutic target to ameliorate the adverse effects of high altitude in adults or of intrauterine hypoxia in the fetus.
据报道,适应高海拔、长期缺氧(LTH)会改变与钾离子通道活性变化相关的脑动脉收缩-舒张反应。我们假设,为了在长期缺氧期间维持氧合,成年绵羊和近足月胎儿的基底动脉(BA)会表现出大电导钙激活钾(BK)通道活性增加。我们分别使用膜片钳电生理学、流式细胞术和共聚焦显微镜,测量了常氧对照、长期缺氧成年和近足月胎儿基底动脉肌细胞中的BK通道活性、表达和细胞表面分布。电生理数据显示,长期缺氧肌细胞中的BK通道表现为:1)钙设定点降低;2)激活电压左移;3)停留时间延长。长期缺氧肌细胞中的BK通道似乎也有更多的去磷酸化。这些差异共同使长期缺氧的BK通道对激活更敏感。流式细胞术研究表明,长期缺氧的胎儿表现出BK β1亚基表面表达增加。此外,在两个胎儿组中,共聚焦显微镜显示BK通道聚集增加,并与肌细胞脂筏共定位。我们得出结论,长期缺氧的基底动脉中BK通道活性增加与通道对钙的亲和力增加和电压激活左移有关。脑血管BK通道活性增加可能是成年和近足月胎儿绵羊适应长期高海拔缺氧的一种机制。我们的研究结果表明,增加脑肌细胞中的BK通道活性可能是改善成年人高海拔或胎儿宫内缺氧不良影响的治疗靶点。