CONFOCAL IMAGING OF CHANGES IN CYTOPLASMIC CA2+ IN GUARD CELLS OF ARABIDOPSIS

拟南芥保卫细胞细胞质 CA2 变化的共聚焦成像

• 批准号: 6282125
• 负责人: GETHYN J ALLEN
• 金额: $ 0.74万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-01 至 1999-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6282125
• 关键词: biomedical resource; environmental health; gas; microscopy; plants; technology /technique

The elevation of cytosolic Ca2+ is a ubiquitous signaling process in eukaryotic cells. Specificity of the physiological response initiated by the Ca2+ signal is in part encoded by the nature of the cytosolic Ca2+ increase. In plant cells, the cell pairs that regulate gaseous exchange in the leaf, specificity to various stimuli (light, temperature, abscisic acid, CO2) is thought to be encoded by spatial, temporal and amplitude (STA) variations in the Ca2+ signal. We plan to dissect these comples Ca2+ signals using single guard cell imaging utilizing the techniques available at NCMIR. Previous studies in the laboratory of Dr. J.I. Schroeder have shown that guard cells can be loaded with the Ca2+ -sensitive dyes Calcium green and Fura 2 by a non-invasive ester loading technique. This significant advance can now be utilized to study Ca2+ signals in these cells. Questions such as the source of Ca2+ (extracellular or intracellular), the STA nature of the signal and the possible oscillation of the signal will be addressed. The information obtained from the images obtained at NCMIR will be combined with concurrent experiments performed in the Schroeder laboratory that utilize electrophysiological techniques such as patch clamping and single electrode voltage clamp. The combination of these techniques will yield important information on the nature and regulation of the transport processes that underlie the generation of Ca2+ signals in guard cells. The work will use Arabidopsis thaliana guard cells. A. thaliana mutants defective in guard cell function have been isolated in the Schroeder laboratory. These mutants will be tested for variations in their CA2+ signaling phenotype, so specific mutations can be linked to defined roles in guard cell signaling. The mutant work will focus particularly on the hormone Absisic acid (ABA). ABA regulates guard cell turgor, and therefore, stomatal aperture by Ca2+-mediated regulation of ion channels. Many mutants are now available in the Schroeder laboratory which show aberrant ABA responses. Using these ABA mutants and an integration of the techniques available at NCMIR and in the Schroeder laboratory could yield important information on the mechanism of action of this important plant hormone. Work on this project is underway.

胞质 Ca2+ 升高是一个普遍存在的信号传导过程 在真核细胞中。 生理反应的特异性 由 Ca2+ 信号引发的部分是由 Ca2+ 信号的性质编码的 胞质Ca2+增加。 在植物细胞中，调节细胞对 叶子中的气体交换，对各种刺激（光、 温度、脱落酸、CO2）被认为是由空间编码的， Ca2+ 信号的时间和幅度 (STA) 变化。 我们计划 使用单保卫细胞成像剖析这些复杂的 Ca2+ 信号 利用 NCMIR 提供的技术。 之前的研究在 J.I.博士实验室施罗德证明保卫细胞可以 负载 Ca2+ 敏感染料钙绿和 Fura 2 非侵入性酯装载技术。 这一重大进步可以 现在可用于研究这些细胞中的 Ca2+ 信号。 诸如此类的问题 作为 Ca2+ 的来源（细胞外或细胞内），STA 性质 信号和信号可能的振荡将是 已解决。 从 NCMIR 获得的图像中获得的信息 将与同时进行的实验相结合 施罗德实验室利用电生理技术，例如 如膜片钳和单电极电压钳。 组合 这些技术将产生有关性质和的重要信息 对产生的运输过程进行监管 保卫细胞中的 Ca2+ 信号。 该作品将使用拟南芥 保卫细胞。 保卫细胞功能缺陷的拟南芥突变体 已被隔离在施罗德实验室。 这些突变体将是 测试了 CA2+ 信号表型的变化，因此具有特异性 突变可能与保卫细胞信号传导中的特定作用有关。 这 突变体工作将特别关注脱落酸（ABA）激素。 ABA 通过调节保卫细胞的膨压，从而调节气孔孔径 Ca2+ 介导的离子通道调节。 现在很多变种人 Schroeder 实验室提供，显示异常 ABA 回应。 使用这些 ABA 突变体和整合 NCMIR 和 Schroeder 实验室现有的技术可以 提供有关该作用机制的重要信息 重要的植物激素。 该项目的工作正在进行中。