Webb Sarah E, 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.
Biochim Biophys Acta. 2012 Aug;1820(8):1160-8. doi: 10.1016/j.bbagen.2011.12.008. Epub 2011 Dec 17.
In recent years, as our understanding of the various roles played by Ca2+ signaling in development and differentiation has expanded, the challenge of imaging Ca2+ dynamics within living cells, tissues, and whole animal systems has been extended to include specific signaling activity in organelles and non-membrane bound sub-cellular domains.
In this review we outline how recent advances in genetics and molecular biology have contributed to improving and developing current bioluminescence-based Ca2+ imaging techniques. Reporters can now be targeted to specific cell types, or indeed organelles or domains within a particular cell.
These advances have contributed to our current understanding of the specificity and heterogeneity of developmental Ca2+ signaling. The improvement in the spatial resolution that results from specifically targeting a Ca2+ reporter has helped to reveal how a ubiquitous signaling messenger like Ca2+ can regulate coincidental but different signaling events within an individual cell; a Ca2+ signaling paradox that until now has been hard to explain.
Techniques used to target specific reporters via genetic means will have applications beyond those of the Ca2+ signaling field, and these will, therefore, make a significant contribution in extending our understanding of the signaling networks that regulate animal development. This article is part of a Special Issue entitled Biochemical, biophysical and genetic approaches to intracellular calcium signalling.
近年来,随着我们对Ca2+信号在发育和分化中所起各种作用的理解不断扩展,对活细胞、组织和整个动物系统内Ca2+动态进行成像的挑战已延伸至包括细胞器和非膜结合亚细胞结构域中的特定信号活性。
在本综述中,我们概述了遗传学和分子生物学的最新进展如何有助于改进和发展当前基于生物发光的Ca2+成像技术。现在,报告基因可以靶向特定的细胞类型,或者实际上是特定细胞内的细胞器或结构域。
这些进展有助于我们目前对发育中Ca2+信号特异性和异质性的理解。通过特异性靶向Ca2+报告基因所带来的空间分辨率的提高,有助于揭示像Ca2+这样普遍存在的信号信使如何在单个细胞内调节同时发生但不同的信号事件;这是一个迄今为止难以解释的Ca2+信号悖论。
通过遗传手段靶向特定报告基因的技术将具有超出Ca2+信号领域的应用,因此,这些技术将在扩展我们对调节动物发育的信号网络的理解方面做出重大贡献。本文是名为《细胞内钙信号传导的生化、生物物理和遗传方法》特刊的一部分。