Webb Sarah E, Chan Ching Man, Miller Andrew L
Division of Life Science and State Key Laboratory of Molecular Neuroscience, The Hong Kong University of Science & Technology, Clear Water Bay, Kowloon, Hong Kong, People's Republic of China.
Cold Spring Harb Protoc. 2013 May 1;2013(5):383-6. doi: 10.1101/pdb.top066316.
Ca(2+) signals, whether transient pulses, propagating waves, or long-duration, steady gradients, are generally considered to play an important role in the pattern-forming events that occur during vertebrate development. One vertebrate that has long been a favorite of embryologists because of its ex utero development and the optical clarity of its embryos is the zebrafish, Danio rerio. Using the bioluminescent Ca(2+) reporter aequorin, distinct Ca(2+) signals have been reported for at least the first 48 h of zebrafish development, with signals becoming progressively more complex as the embryo develops. Here we provide a general introduction to aequorin and its use in monitoring Ca(2+) signals and discuss methods for introducing aequorin into zebrafish embryos.
钙离子信号，无论是瞬态脉冲、传播波还是长时间的稳定梯度，通常被认为在脊椎动物发育过程中发生的模式形成事件中起着重要作用。由于其体外发育和胚胎的光学清晰度，长期以来一直是胚胎学家喜爱的一种脊椎动物是斑马鱼（Danio rerio）。使用生物发光钙离子报告蛋白水母发光蛋白，已报道斑马鱼发育的至少前48小时存在不同的钙离子信号，随着胚胎发育，信号逐渐变得更加复杂。在这里，我们对水母发光蛋白及其在监测钙离子信号中的应用进行了一般性介绍，并讨论了将水母发光蛋白引入斑马鱼胚胎的方法。
