CIC-3 chloride ion channels in vascular smooth muscle

血管平滑肌中的CIC-3氯离子通道

• 批准号: 7216399
• 负责人: FRED S LAMB
• 金额: $ 27.97万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7216399
• 关键词: Acids; Affect; Agonist; Alternative Splicing; Amino Acid Motifs; Angiotensin II; Animals; Anions; Antibiotic A23187; Antioxidants; Aorta; Applications Grants; Arteries; Atherosclerosis; Binding; Biochemical; Biological; Biological Assay; Blindness; Blood Pressure; Blood Vessels; Calcium; Calcium/calmodulin-dependent protein kinase; Cardiovascular Abnormalities; Cardiovascular Diseases; Cardiovascular Physiology; Cardiovascular system; Catecholamines; Cell membrane; Cell surface; Cells; Cessation of life; Characteristics; Chloride Channels; Chloride Ion; Chlorides; Chronic; ClC-3 channel; Complement component C1s; Complex; Condition; Conflict (Psychology); Coupled; Cultured Cells; Cystic Fibrosis Transmembrane Conductance Regulator; Cytochromes; Cytoplasm; Data; Defect; Depth; Dose; Endosomes; Endothelium; Epitopes; Event; Exons; Exposure to; Family; Feline Immunodeficiency Virus; Functional disorder; GTP-Binding Proteins; Genes; Green Fluorescent Proteins; Growth; Heterogeneity; High Blood Pressure; Hippocampus (Brain); Human; Hypertension; In Vitro; Intervention; Ionophores; Ions; Knock-out; Knockout Mice; Knowledge; Laboratories; Left; Left Ventricular Hypertrophy; Link; Literature; Localized; Location; Luciferases; Measures; Mediating; Membrane; Membrane Protein Traffic; Mesentery; Metabolism; Modeling; Movement; Mus; Mutation; Nature; Nitric Oxide; Nitroprusside; Oxidative Stress; Pathway interactions; Phenotype; Physical activity; Physiological; Play; Process; Production; Protein Binding; Protein Isoforms; Proteins; RNA Splicing; Reactive Oxygen Species; Recombinant Proteins; Recombinants; Regulation; Relaxation; Resistance; Rest; Role; Screening procedure; Seizures; Series; Signal Pathway; Signal Transduction; Single-Gene Defect; Smooth Muscle Myocytes; Superoxides; Swelling; Synaptic Vesicles; System; Tachycardia; Testing; Time; Tissues; Transcript; Variant; Vascular Smooth Muscle; Vasodilation; Vasodilation disorder; Vesicle; Viral; Yeasts; base; concept; erection; extracellular; insight; member; novel; patch clamp; promoter; protein expression; protein protein interaction; protein structure; receptor; research study; response; retinal neuron; sex; trafficking; ventricular hypertrophy; voltage; yeast two hybrid system

Chloride (CI) currents contribute to agonist-induced depolarization of vascular smooth muscle (VSM) cells. As part of the original grant proposal for which this application is a continuation, we created a mouse lacking a specific chloride channel, CIC-3 (encoded by the CIcn3 gene). Five findings from our laboratory demonstrate the importance of CIC-3 to cardiovascular function: 1) Animals lacking CIC-3 CI channels display multiple cardiovascular abnormalities including; hypertension, left ventricular hypertrophy and diastolic dysfunction, and impaired endothelium-dependent relaxation in resistance vessels, 2) CIC-3 channels are located intracellularly in resting VSM cells, but are inserted into the plasma membrane in response to All, 3) there is an unappreciated diversity of CIC-3 protein structure resulting from alternative splicing that may alter membrane trafficking, 4) Clcn3-/- cells have increased levels of intracellular reactive oxygen species (ROS), and 5) superoxide anion may pass through CIC-3 channels. There is conflicting data in the literature as to the biophysical nature of CIC-3. These inconsistencies may reflect a lack of in-depth knowledge of channel localization within the cell and the mechanisms that move the channel between cellular compartments. We will first carefully define the localization and trafficking of CIC-3. We will then test the hypothesis that the altered microvascular function observed in Clcn3-/- mice is related to the absence of a conductance that normally provides a mechanism by which superoxide anion moves across biological membranes. In this renewal application, we will; 1) Define the subcellular localization of native CIC-3 protein in murine VSM and identify factors that regulate the trafficking of CIC-3 to the plasma membrane in response to angiotensin II, 2) Define the subcellular localization of the six distinct splice variants of CIC-3 in VSM cells using FIV-driven expression of recombinant CIC-3 protein and identify motifs and physiologic factors that regulate membrane trafficking of CIC-3, and 3) Determine why intracellular ROS levels are elevated in Clcn3-/- cells and tissues and discern if this increase is physiologically relevant. The CIC-3 knockout mouse represents a novel single-gene defect model of hypertension. Careful analysis of the physiological defects in Clcn3-/- mice will yield important insight into cellular ROS metabolism and the link between ROS and high blood pressure.

氯化物 (CI) 电流有助于激动剂诱导的血管平滑肌 (VSM) 细胞去极化。 作为本申请的延续的原始资助提案的一部分，我们创造了一种缺乏特定氯离子通道 CIC-3（由 CIcn3 基因编码）的小鼠。 我们实验室的五项发现证明了 CIC-3 对心血管功能的重要性：1) 缺乏 CIC-3 CI 通道的动物表现出多种心血管异常，包括；高血压、左心室肥大和舒张功能障碍，以及阻力血管中内皮依赖性舒张受损，2) CIC-3 通道位于静息 VSM 细胞的细胞内，但响应 All 插入到质膜中，3) 由于可能改变膜的选择性剪接，CIC-3 蛋白质结构存在未被认识到的多样性 运输，4) Clcn3-/- 细胞细胞内活性氧 (ROS) 水平增加，5) 超氧阴离子可能通过 CIC-3 通道。 关于 CIC-3 的生物物理性质，文献中存在相互矛盾的数据。 这些不一致可能反映出缺乏对细胞内通道定位以及在细胞区室之间移动通道的机制的深入了解。 我们将首先仔细定义 CIC-3 的本地化和贩运。 然后，我们将测试以下假设：在 Clcn3-/- 小鼠中观察到的微血管功能改变与电导的缺失有关，而电导通常提供超氧阴离子跨生物膜移动的机制。 在此续签申请中，我们将； 1) 定义小鼠 VSM 中天然 CIC-3 蛋白的亚细胞定位，并识别响应血管紧张素 II 调节 CIC-3 运输至质膜的因素，2) 使用 FIV 驱动的重组 CIC-3 蛋白表达，定义 VSM 细胞中 CIC-3 六种不同剪接变体的亚细胞定位，并识别基序和生理因素 调节 CIC-3 的膜运输，以及 3) 确定 Clcn3-/- 细胞和组织中细胞内 ROS 水平升高的原因，并辨别这种升高是否具有生理相关性。 CIC-3 基因敲除小鼠代表了一种新型的单基因缺陷高血压模型。 仔细分析 Clcn3-/- 小鼠的生理缺陷将有助于深入了解细胞 ROS 代谢以及 ROS 与高血压之间的联系。

项目成果

Endothelial superoxide production is altered in sheep programmed by early gestation dexamethasone exposure.

妊娠早期接触地塞米松会改变绵羊内皮超氧化物的产生。

• DOI: 10.1159/000105521
• 发表时间: 2008
• 期刊: Neonatology
• 影响因子: 2.5
• 作者: Roghair,RobertD;MillerJr,FrancisJ;Scholz,ThomasD;Lamb,FredS;Segar,JeffreyL
• 通讯作者: Segar,JeffreyL

Impact of maternal dexamethasone on coronary PGE(2) production and prostaglandin-dependent coronary reactivity.

母体地塞米松对冠状动脉 PGE(2) 产生和前列腺素依赖性冠状动脉反应性的影响。

• DOI: 10.1152/ajpregu.00658.2011
• 发表时间: 2012
• 期刊: American journal of physiology. Regulatory, integrative and comparative physiology
• 作者: Roghair,RobertD;Volk,KennethA;Lamb,FredS;Segar,JeffreyL
• 通讯作者: Segar,JeffreyL

Newborn lamb coronary artery reactivity is programmed by early gestation dexamethasone before the onset of systemic hypertension.

• DOI: 10.1152/ajpregu.00369.2005
• 发表时间: 2005-10
• 期刊: American journal of physiology. Regulatory, integrative and comparative physiology
• 作者: R. Roghair;J. Segar;Ram V. Sharma;M. Zimmerman;D. Jagadeesha;E. Segar;T. Scholz;F. Lamb

Vascular nitric oxide and superoxide anion contribute to sex-specific programmed cardiovascular physiology in mice.

血管一氧化氮和超氧阴离子有助于小鼠性别特异性的程序性心血管生理学。

• DOI: 10.1152/ajpregu.90756.2008
• 发表时间: 2009
• 期刊: American journal of physiology. Regulatory, integrative and comparative physiology
• 作者: Roghair,RobertD;Segar,JeffreyL;Volk,KennethA;Chapleau,MarkW;Dallas,LindsayM;Sorenson,AnnaR;Scholz,ThomasD;Lamb,FredS
• 通讯作者: Lamb,FredS

Early gestation dexamethasone alters baroreflex and vascular responses in newborn lambs before hypertension.

妊娠早期地塞米松在高血压之前改变新生羔羊的压力反射和血管反应。

• DOI: 10.1152/ajpregu.00677.2005
• 发表时间: 2006
• 期刊: American journal of physiology. Regulatory, integrative and comparative physiology
• 作者: Segar,JeffreyL;Roghair,RobertD;Segar,EmilyM;Bailey,MelissaC;Scholz,ThomasD;Lamb,FredS
• 通讯作者: Lamb,FredS