Application of single-cell microfluorimetry to neurotoxicology assays. 2009

Tobi L Limke, and William D Atchison
Millipore Corporation, Billerica, Massachusetts, USA.

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.

UI MeSH Term Description Entries
D009474 Neurons The basic cellular units of nervous tissue. Each neuron consists of a body, an axon, and dendrites. Their purpose is to receive, conduct, and transmit impulses in the NERVOUS SYSTEM. Nerve Cells,Cell, Nerve,Cells, Nerve,Nerve Cell,Neuron
D002118 Calcium A basic element found in nearly all tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes. Coagulation Factor IV,Factor IV,Blood Coagulation Factor IV,Calcium-40,Calcium 40,Factor IV, Coagulation
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D003592 Cytophotometry A method for the study of certain organic compounds within cells, in situ, by measuring the light intensities of the selectively stained areas of cytoplasm. The compounds studied and their locations in the cells are made to fluoresce and are observed under a microscope. Microfluorometry,Cytophotometries,Microfluorometries
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013050 Spectrometry, Fluorescence Measurement of the intensity and quality of fluorescence. Fluorescence Spectrophotometry,Fluorescence Spectroscopy,Spectrofluorometry,Fluorescence Spectrometry,Spectrophotometry, Fluorescence,Spectroscopy, Fluorescence
D014116 Toxicology The science concerned with the detection, chemical composition, and biological action of toxic substances or poisons and the treatment and prevention of toxic manifestations. Toxinology,Evidence Based Toxicology,Evidence-Based Toxicology,Based Toxicologies, Evidence,Based Toxicology, Evidence,Evidence Based Toxicologies,Evidence-Based Toxicologies,Toxicologies, Evidence Based,Toxicologies, Evidence-Based,Toxicology, Evidence Based,Toxicology, Evidence-Based
D053078 Membrane Potential, Mitochondrial The voltage difference, normally maintained at approximately -180mV, across the INNER MITOCHONDRIAL MEMBRANE, by a net movement of positive charge across the membrane. It is a major component of the PROTON MOTIVE FORCE in MITOCHONDRIA used to drive the synthesis of ATP. Delta Psi M,DeltaPsi M,DeltapsiM,Mitochondrial Membrane Potential,Mitochondrial Transmembrane Potential,M, DeltaPsi,Membrane Potentials, Mitochondrial,Mitochondrial Membrane Potentials,Mitochondrial Transmembrane Potentials,Transmembrane Potential, Mitochondrial,Transmembrane Potentials, Mitochondrial
D059010 Single-Cell Analysis Assaying the products of or monitoring various biochemical processes and reactions in an individual cell. Analyses, Single-Cell,Analysis, Single-Cell,Single Cell Analysis,Single-Cell Analyses

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