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首页> 外文会议>Biomedical Imaging: Reporters, Dyes, and Instrumentation >Intracellular pH (pHin) and cytosolic calcium (Ca2+cyt) regulation via ATPases: studies in cell populations single cells and subcellular compartments
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Intracellular pH (pHin) and cytosolic calcium (Ca2+cyt) regulation via ATPases: studies in cell populations single cells and subcellular compartments

机译：通过ATPase调节细胞内pH（pHin）和胞质钙（Ca2 + cyt）：在细胞群体中的单细胞和亚细胞区室研究

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Abstract: Changes in pH$+in$/ and (Ca$+2$PLU$/)$+cyt$/ are important in the signal transduction mechanisms leading to many physiological responses including cell growth, motility, secretion/exocytosis, etc. The concentrations of these ions are regulated via primary and secondary ion transporting mechanisms. In diabetes, specific pH and Ca$+2$PLU$/ regulatory mechanism might be altered. To study these ions, we employ fluorescence spectroscopy, and cell imagin spectroscopy/confocal microscopy. pH and Ca$+2$PLU$/ indicators are loaded in the cytosol with acetoxymethyl ester forms of dyes, and in endosomal/lysosomal (E/L) compartments by overnight incubation of cells with dextran- conjugated ion fluorescent probes. We focus on specific pH and Ca$+2$PLU$/ regulatory systems: plasmalemmal vacuolar- type H$+$PLU$/-ATPases (pm V-ATPases) and sarcoplasmic/endoplasmic reticulum Ca$+2$PLU$/-ATPases (SERCA). As experimental models, we employ vascular smooth muscle (VSM) and microvascular endothelial cells. We have chosen these cells because they are important in blood flow regulation and in angiogenesis. These processes are altered in diabetes. In many cell types, ion transport processes are dependent on metabolism of glucose for maximal activity. Our main findings are: (a) glycolysis coupling the activity of SERCA is required for cytosolic Ca$+2$PLU$/ homeostasis in both VSM and microvascular endothelial cells; (b) E/L compartments are important for pH and Ca$+2$PLU$/ regulation via H$+$PLU$/-ATPases and SERCA, respectively; and (c) pm-V- ATPases are important for pH$+in$/ regulation in microvascular endothelial cells. !48
机译：摘要：pH $ + in $ /和（Ca $ + 2 $ PLU $ /）$ + cyt $ /的变化在信号转导机制中很重要，导致许多生理反应，包括细胞生长，运动，分泌/胞吐等。这些离子的浓度通过一级和二级离子传输机制进行调节。在糖尿病中，特定的pH和Ca $ + 2 $ PLU $ /调节机制可能会改变。为了研究这些离子，我们采用了荧光光谱和细胞成像光谱/共聚焦显微镜。 pH和Ca $ + 2 $ PLU $ /指示剂通过与葡聚糖偶联的离子荧光探针一起孵育过夜，在细胞溶质中装载有乙酰氧基甲基酯形式的染料，并在内体/溶酶体（E / L）隔室中装载。我们关注特定的pH和Ca $ + 2 $ PLU $ /调节系统：质膜液泡型H $ + $ PLU $ /-ATPases（pm V-ATPases）和肌浆/内质网Ca $ + 2 $ PLU $ /- ATPase（SERCA）。作为实验模型，我们采用血管平滑肌（VSM）和微血管内皮细胞。我们选择这些细胞是因为它们在血流调节和血管生成中很重要。这些过程在糖尿病中有所改变。在许多细胞类型中，离子转运过程依赖于葡萄糖的代谢才能发挥最大活性。我们的主要发现是：（a）在VSM和微血管内皮细胞中，胞浆Ca $ + 2 $ PLU $ /稳态需要糖酵解结合SERCA的活性； （b）E / L区隔对于分别通过H $ + $ PLU $ /-ATPases和SERCA调节pH和Ca $ + 2 $ PLU $ /很重要； （c）pm-V- ATPase对微血管内皮细胞的pH $ + in $ /调节很重要。 ！48

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