Role of TRP Channels on Collecting Duct Calcium Dynamics
TRP 通道对集合管钙动态的作用
基本信息
- 批准号:7046054
- 负责人:
- 金额:$ 28.71万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2005
- 资助国家:美国
- 起止时间:2005-04-01 至 2010-03-31
- 项目状态:已结题
- 来源:
- 关键词:calcium channelcalcium fluxcalcium ioncell lineconfocal scanning microscopyimmunofluorescence techniqueimmunoprecipitationlaboratory mousemechanical stressmembrane permeabilitymolecular /cellular imagingnorthern blottingsphospholipase Cpolymerase chain reactionprotein isoformsprotein kinaseprotein localizationprotein structure functionprotein transportrenal tubular transportrenal tubulesmall interfering RNAtissue /cell culturevoltage /patch clampwestern blottings
项目摘要
DESCRIPTION (provided by applicant):The overall goal of this proposal is to elucidate the role of Ca-permeable TRPV channel isoforms in regulating mechanically-induced intracellular Ca, [Ca]i, dynamics and Ca reabsorption in renal cortical collecting duct. Mechanical stresses, such as fluid shear stress or hypoosmotic stress, vary widely in late distal tubule and cortical collecting duct (CCD) especially during pathophysiological states (e.g., hyponatremia, diabetes insipidus, diabetes mellitus, diuretic administration), which will contribute to hypocaliuric and/or hypercaliuric states that lead to altered Ca balance. We have implicated two TRPV channels as being mechanosensitive and central to regulating mechano-stimulated Ca entry and reabsorption in mouse CCD. The following four specific aims are proposed to achieve the overall goal: 1. To determine the relative role and pharmacological/ electrophysiological properties of mechanoregulated Ca entry pathways (Ca imaging, patch clamp) in control of [Ca]i dynamics in mouse CCD cells; 2. To identify TRPV channel isoforms expressed in mouse CCD cells (RT-PCR, Northern blot, immunofluorescence, immunoblots) and to determine which specific isoforms participate in mechanoregulated [Ca] dynamics; 3. To delineate the relative contribution of specific mechanoregulated TRPV channels in Ca reabsorption in mouse CCD cells (overexpression/siRNA knockdown) and to examine the mechanism of regulation of the channels (phospholipases, protein kinases); and 4. To evaluate the role of membrane localization/trafficking of TRPV isoforms (confocal microscopy/immunofluorescence, biotinylation-streptavidin purification) and the association with accessory proteins (immunoprecipitation/pull-downs), including other TRPs, on mechanoregulated [Ca]i dynamics and Ca reabsorption. The outcome of the study has important health relatedness as it will provide new insights into the poorly understood mechanisms of calcium signaling and reabsorption in renal tubules and provide a critical new understanding of the mechanism by which altered pathophysiological conditions regulate calcium reabsorption in renal cells.
描述(由申请人提供):该提案的总体目标是阐明可渗透性的TRPV通道同工型在调节机械诱导的细胞内CA中的作用,[CA] I,动力学和Ca在肾脏皮质收集管中的吸收。 Mechanical stresses, such as fluid shear stress or hypoosmotic stress, vary widely in late distal tubule and cortical collecting duct (CCD) especially during pathophysiological states (e.g., hyponatremia, diabetes insipidus, diabetes mellitus, diuretic administration), which will contribute to hypocaliuric and/or hypercaliuric states that lead to altered Ca balance.我们暗示了两个TRPV通道是机械敏感的,并且是调节小鼠CCD中的机械刺激的Ca进入和重吸收的核心。提出了以下四个特定目标来实现总体目标:1。确定机制调节的Ca进入途径(CA成像,贴片夹)在控制小鼠CCD细胞中[CA] I动力学中的相对作用和药理/电生理特性; 2。识别在小鼠CCD细胞中表达的TRPV通道同工型(RT-PCR,北印迹,免疫荧光,免疫印迹),并确定哪些特定同工型参与机械调节的[CA]动力学; 3。描绘小鼠CCD细胞中CA重吸收中特定机械调节的TRPV通道的相对贡献(过表达/siRNA敲低),并检查通道调节的机理(磷脂酶,蛋白质激酶);和4。评估TRPV同工型(共聚焦显微镜/免疫荧光,生物素化 - 链霉亲和素纯化)的膜定位/运输的作用,并与辅助蛋白(包括免疫沉淀/拉力下降)相关联,包括其他TRP,包括其他机械化[CA] INTYRAPTION和CA REANTION和CA REAMETICT和CA REABERIPTION。该研究的结果具有重要的健康相关性,因为它将为肾小管中钙信号传导和重吸收的机制提供新的见解,并对改变病理生理状况调节肾细胞中钙化的机制有一个关键的新理解。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
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ROGER Gordon O'NEIL其他文献
ROGER Gordon O'NEIL的其他文献
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{{ truncateString('ROGER Gordon O'NEIL', 18)}}的其他基金
RNA Aptamers Selective for TPRV Channels
针对 TPRV 通道的选择性 RNA 适体
- 批准号:
7465745 - 财政年份:2008
- 资助金额:
$ 28.71万 - 项目类别:
RNA Aptamers Selective for TPRV Channels
针对 TPRV 通道的选择性 RNA 适体
- 批准号:
7656649 - 财政年份:2008
- 资助金额:
$ 28.71万 - 项目类别:
VALIDATION OF FLUORESCENT DEOXYGLUCOSE (2-NBDG) IN TUMORS
荧光脱氧葡萄糖 (2-NBDG) 在肿瘤中的验证
- 批准号:
7053173 - 财政年份:2006
- 资助金额:
$ 28.71万 - 项目类别:
Role of TRP Channels on Collecting Duct Calcium Dynamics
TRP 通道对集合管钙动态的作用
- 批准号:
6907669 - 财政年份:2005
- 资助金额:
$ 28.71万 - 项目类别:
Role of TRP Channels on Collecting Duct Calcium Dynamics
TRP 通道对集合管钙动态的作用
- 批准号:
7215599 - 财政年份:2005
- 资助金额:
$ 28.71万 - 项目类别:
Role of TRP Channels on Collecting Duct Calcium Dynamics
TRP 通道对集合管钙动态的作用
- 批准号:
7590356 - 财政年份:2005
- 资助金额:
$ 28.71万 - 项目类别:
Role of TRP Channels on Collecting Duct Calcium Dynamics
TRP 通道对集合管钙动态的作用
- 批准号:
7390360 - 财政年份:2005
- 资助金额:
$ 28.71万 - 项目类别:
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