Analyses of Regulatory Mechanism of Na+-K+-Cl- Cotransporter Function in Wild Type and Mutant
野生型和突变体Na-K-Cl-协同转运蛋白功能调控机制分析
基本信息
- 批准号:17591939
- 负责人:
- 金额:$ 2.11万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:2005
- 资助国家:日本
- 起止时间:2005 至 2006
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
The results obtained in the research from April in 2005 to March in 2006 are as follows :1. Establishment of culture cell system for analyses of Cl transportForskolin, a cAMP-increasing agent, induced an ionic current in MDCK and Calu-3 cells, which have an intrinsic Na^+-K^+-2Cl^- cotransporter activity. HEK293 cells were stained by anti-human Na^+-K^+-2Cl^- cotransporter but not by anti-rat Na^+-K^+-2Cl^- cotransporter, while the cells were stained by anti-rat Na^+-K^+-2Cl^- cotransporter after transfection with rat Na^+-K^+-2Cl^- cotransporter-containing plasmid. We started mutation experiments of the Na^+-K^+-2Cl^- cotransporter.2. Analyses of the rat Na^+-K^+-2Cl^- cotransporter activity in salivary glandsTo clarify the regulation mechanism of Na^+-K^+-2Cl^- cotransporter activity by Ca^<2+> and cAMP signals, we investigated the effects of β-adrenergic receptor stimulation on carbachol (CCh)-induced Cl^- current and the effect of α and β receptor stimulation by noradrenaline (NA). … More CCh induced a bumetanide-sensitive oscillatory inward Cl^- current, suggesting that the current carrier is Cl^- which moves into the cell through the Na^+-K^+-2Cl^- cotransporter. Addition of the β-adrenergic agonist, isoproterenol, and forskolin + IBMX suppressed the CCh-induced Cl^- current. This suggests that the reduction of Na^+-K^+-2Cl^- cotransporter activity by cAMP may results in decrease in electrolyte transport and then suppression of salivation during muscarinic receptor stimulation by β-adrenergic stimulation. NA induced an oscillatory anion current of which amplitude gradually decreased to a steady-state level in submandibular acinar cells. The β-adrenergic antagonist, propranolol, did not induce any current, but inhibited the gradual decrease in the NA-induced current, suggesting that the current is induced via the α-adrenergic stimulation by NA and suppressed via the β-adrenergic stimulation by NA and that the relatively small volume of saliva induced by sympathetic stimulation may be at least partially due to the reduction of Cl secretion in the acinar cells by the β-adrenergic stimulation. Less
2005年4月至2006年3月的研究结果如下:1。培养细胞体系的建立camp增加剂forskolin在具有Na^+- k ^+- 2cl ^-共转运体活性的MDCK和Calu-3细胞中诱导离子电流。HEK293细胞用抗人Na^+- k ^+- 2cl ^-共转运体染色,不用抗大鼠Na^+- k ^+- 2cl ^-共转运体染色,用含Na^+- k ^+- 2cl ^-共转运体的大鼠质粒转染细胞后,用抗大鼠Na^+- k ^+- 2cl ^-共转运体染色。我们开始了Na^+- k ^+- 2cl ^-共转运体的突变实验。为了阐明Ca^<2+>和cAMP信号对Na^+- k ^+- 2cl ^-共转运体活性的调控机制,我们研究了β-肾上腺素受体刺激对碳bachol (CCh)诱导的Cl^-电流的影响以及去甲肾上腺素(Na)刺激α和β受体的作用。更多的CCh诱导了对布美他尼敏感的向内振荡Cl^-电流,表明电流载体是Cl^-,它通过Na^+- k ^+- 2cl ^-共转运体进入细胞。β-肾上腺素能激动剂、异丙肾上腺素和福斯克林+ IBMX的加入抑制了cch诱导的Cl^-电流。这表明cAMP降低Na^+- k ^+- 2cl ^-共转运体活性可能导致β-肾上腺素能刺激毒蕈碱受体时电解质转运减少,进而抑制唾液分泌。NA在颌下腺泡细胞中诱导振荡阴离子电流,其振幅逐渐下降至稳态水平。β-肾上腺素能拮抗剂心得安不诱导任何电流,但能抑制NA诱导电流的逐渐减少,提示NA通过α-肾上腺素能刺激诱导电流,而NA通过β-肾上腺素能刺激抑制电流,交感刺激诱导的相对小的唾液量可能至少部分是由于β-肾上腺素能刺激减少了腺泡细胞的Cl分泌。少
项目成果
期刊论文数量(6)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Electrolyte transport and evaluation of secretory function in salivary glands.
唾液腺电解质转运和分泌功能评估。
- DOI:
- 发表时间:2006
- 期刊:
- 影响因子:0
- 作者:Hirono;C.
- 通讯作者:C.
Variousness of carbachol-induced bicarbonate current in rat parotid duct cells.
大鼠腮腺管细胞中卡巴胆碱诱导的碳酸氢盐电流的多样性。
- DOI:
- 发表时间:2005
- 期刊:
- 影响因子:0
- 作者:Hirono;C.
- 通讯作者:C.
カルバコール刺激でラット耳下腺腺房細胞から分泌されるC1^-電流のイソプロテレノールおよびフォルスコリンによる抑制
异丙肾上腺素和毛喉素刺激卡巴胆碱对大鼠腮腺腺泡细胞分泌的 C1^-电流的抑制
- DOI:
- 发表时间:2006
- 期刊:
- 影响因子:0
- 作者:広野 力
- 通讯作者:広野 力
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