Limke Tobi L, Atchison William D
Millipore Corporation, Billerica, Massachusetts, USA.
Curr Protoc Toxicol. 2009 Nov;Chapter 12:Unit 12.15. doi: 10.1002/0471140856.tx1215s42.
Intracellular signaling events play fundamental roles in regulating physiological function. In neurons, these include inducing growth and differentiation, secretion, gene expression, and controlling processes associated with learning and memory. All of these processes have in common the vital dependence on changes in intracellular Ca²(+) Ca²(+). Numerous toxicants, including metals, polychlorinated biphenyls, and biological neurotoxins, can disrupt Ca²(+). Understanding how toxicants disrupt Ca²(+)-dependent neuronal signaling, and thus induce neuronal death or dysfunction, requires the ability to monitor Ca²(+) at the level of individual cells. A series of fluorophores that can report on changes in Ca²(+) has been pivotal in this process. This section describes how to use these fluorophores to study effects of neurotoxicants on two types of processes: changes in Ca²(+) in individual cells and changes in mitochondrial membrane potential. Similar techniques using distinct fluorophores can be applied to other physiological processes.
细胞内信号事件在调节生理功能中发挥着基础作用。在神经元中，这些作用包括诱导生长与分化、分泌、基因表达以及控制与学习和记忆相关的过程。所有这些过程的共同之处在于对细胞内Ca²⁺（[Ca²⁺]i）变化的至关重要的依赖性。许多有毒物质，包括金属、多氯联苯和生物神经毒素，都能扰乱[Ca²⁺]i。要了解有毒物质如何扰乱依赖Ca²⁺的神经元信号传导，进而导致神经元死亡或功能障碍，就需要具备在单个细胞水平监测[Ca²⁺]i的能力。一系列能够报告[Ca²⁺]i变化的荧光团在这一过程中起到了关键作用。本节描述了如何使用这些荧光团来研究神经毒素对两种类型过程的影响：单个细胞内[Ca²⁺]i的变化以及线粒体膜电位的变化。使用不同荧光团的类似技术可应用于其他生理过程。
