Application of single-cell microfluorimetry to neurotoxicology assays.
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(+)](i). Numerous toxicants, including metals, polychlorinated biphenyls, and biological neurotoxins, can disrupt [Ca²(+)](i). Understanding how toxicants disrupt Ca²(+)-dependent neuronal signaling, and thus induce neuronal death or dysfunction, requires the ability to monitor [Ca²(+)](i) at the level of individual cells. A series of fluorophores that can report on changes in [Ca²(+)](i) 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(+)](i) in individual cells and changes in mitochondrial membrane potential. Similar techniques using distinct fluorophores can be applied to other physiological processes.
Millipore Corporation, Billerica, Massachusetts, USA.
Tobi L
TL
Limke
William D
WD
Atchison
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D003592:Cytophotometry / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D059010:Single-Cell Analysis / Q000295:instrumentation / Q000379:methods*; D013050:Spectrometry, Fluorescence; D014116:Toxicology / Q000295:instrumentation / Q000379:methods*
D002118:Calcium