Rosay P, Davies S A, Yu Y, Sözen M A, Kaiser K, Dow J A
Division of Molecular Genetics, Institute of Biomedical and Life Sciences, University of Glasgow, UK.
J Cell Sci. 1997 Aug;110 ( Pt 15):1683-92. doi: 10.1242/jcs.110.15.1683.
Calcium is a ubiquitous second messenger that plays a critical role in both excitable and non-excitable cells. Calcium mobilisation in identified cell types within an intact renal epithelium, the Drosophila melanogaster Malpighian tubule, was studied by GAL4-directed expression of an aequorin transgene. CAP2b, a cardioactive neuropeptide that stimulates fluid secretion by a mechanism involving nitric oxide, causes a rapid, dose-dependent rise in cytosolic calcium in only a single, genetically-defined, set of 77 principal cells in the main (secretory) segment of the tubule. In the absence of external calcium, the CAP2b-induced calcium response is abolished. In Ca2+-free medium, the endoplasmic reticulum Ca2+-ATPase inhibitor, thapsigargin, elevates [Ca2+]i only in the smaller stellate cells, suggesting that principal cells do not contain a thapsigargin-sensitive intracellular pool. Assays for epithelial function confirm that calcium entry is essential for CAP2b to induce a physiological response in the whole organ. Furthermore, the data suggest a role for calcium signalling in the modulation of the nitric oxide signalling pathway in this epithelium. The GAL4-targeting system allows general application to studies of cell-signalling and pharmacology that does not rely on invasive or cytotoxic techniques.
钙是一种普遍存在的第二信使,在可兴奋细胞和非可兴奋细胞中均发挥关键作用。通过GAL4指导的水母发光蛋白转基因表达,研究了完整肾上皮(果蝇黑腹果蝇马氏管)中特定细胞类型的钙动员情况。CAP2b是一种心脏活性神经肽,通过一种涉及一氧化氮的机制刺激液体分泌,仅在肾小管主要(分泌)段中一组由基因定义的77个主细胞中引起细胞溶质钙的快速、剂量依赖性升高。在没有外部钙的情况下,CAP2b诱导的钙反应被消除。在无钙培养基中,内质网钙ATP酶抑制剂毒胡萝卜素仅在较小的星状细胞中升高[Ca2+]i,这表明主细胞中不含有对毒胡萝卜素敏感的细胞内钙库。上皮功能检测证实,钙内流对于CAP2b在整个器官中诱导生理反应至关重要。此外,数据表明钙信号在此上皮中一氧化氮信号通路的调节中发挥作用。GAL4靶向系统允许普遍应用于不依赖侵入性或细胞毒性技术的细胞信号传导和药理学研究。