Speake Tracey, Kibble Jonathan D, Brown Peter D
School of Biological Sciences, University of Manchester, Manchester M13 9PT, United Kingdom.
Am J Physiol Cell Physiol. 2004 Mar;286(3):C611-20. doi: 10.1152/ajpcell.00292.2003. Epub 2003 Nov 5.
The choroid plexuses secrete, and maintain the composition of, the cerebrospinal fluid. K+ channels play an important role in these processes. In this study the molecular identity and properties of the delayed-rectifying K+ (Kv) conductance in rat choroid plexus epithelial cells were investigated. Whole cell K+ currents were significantly reduced by 10 nM dendrotoxin-K and 1 nM margatoxin, which are specific inhibitors of Kv1.1 and Kv1.3 channels, respectively. A combination of dendrotoxin-K and margatoxin caused a depolarization of the membrane potential in current-clamp experiments. Western blot analysis indicated the presence of Kv1.1 and Kv1.3 proteins in the choroid plexus. Furthermore, the Kv1.3 and Kv1.1 proteins appear to be expressed in the apical membrane of the epithelial cells in immunocytochemical studies. The Kv conductance was inhibited by 1 microM serotonin (5-HT), with maximum inhibition to 48% of control occurring in 8 min (P < 0.05 by Student's t-test for paired data). Channel inhibition by 5-HT was prevented by the 5-HT2C antagonist mesulergine (300 nM). It was also attenuated in the presence of calphostin C (a protein kinase C inhibitor). The conductance was partially inhibited by 1,2-dioctanoyl-sn-glycerol and phorbol 12-myristate 13-acetate, both of which activate protein kinase C. These data suggest that 5-HT acts at 5-HT2C receptors to activate protein kinase C, which inhibits the Kv channels. In conclusion, Kv1.1 and Kv1.3 channels make a significant contribution to K+ efflux at the apical membrane of the choroid plexus.
脉络丛分泌并维持脑脊液的成分。钾离子通道在这些过程中发挥着重要作用。在本研究中,对大鼠脉络丛上皮细胞中延迟整流钾离子(Kv)电导的分子特性和性质进行了研究。10 nM的树眼镜蛇毒素-K和1 nM的玛格毒素分别显著降低了全细胞钾离子电流,它们分别是Kv1.1和Kv1.3通道的特异性抑制剂。在电流钳实验中,树眼镜蛇毒素-K和玛格毒素的组合导致膜电位去极化。蛋白质印迹分析表明脉络丛中存在Kv1.1和Kv1.3蛋白。此外,在免疫细胞化学研究中,Kv1.3和Kv1.1蛋白似乎在上皮细胞的顶端膜中表达。Kv电导被1 μM的5-羟色胺(5-HT)抑制,在8分钟内最大抑制至对照的48%(配对数据的Student t检验,P < 0.05)。5-HT2C拮抗剂美舒麦角(300 nM)可防止5-HT对通道的抑制。在钙泊三醇C(一种蛋白激酶C抑制剂)存在的情况下,这种抑制作用也会减弱。1,2-二辛酰基-sn-甘油和佛波酯12-肉豆蔻酸酯13-乙酸酯均可部分抑制该电导,二者均可激活蛋白激酶C。这些数据表明,5-HT作用于5-HT2C受体以激活蛋白激酶C,进而抑制Kv通道。总之,Kv1.1和Kv1.3通道对脉络丛顶端膜的钾离子外流有显著贡献。