Abstract
Intracellular Ca2+ is involved in control of a large variety of cell functions including apoptosis and neuron cell death . For example, intracellular Ca2+ overload is critical in neuron cell death induced by excitotoxicity . Thus, single cell monitoring of intracellular Ca2+ concentration ([Ca2+]cyt ) in neurons concurrently with apoptosis and neuron cell death is widely required.
Procedures for culture and preparation of primary cultures of hippocampal rat neurons and fluorescence imaging of cytosolic Ca2+ concentration in Fura2/AM -loaded neurons are described. We also describe a method for apoptosis detection by immunofluorescence imaging . Finally, a simple method for concurrent measurements of [Ca2+]cyt and apoptosis in the same neurons is described.
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Acknowledgments
This work was supported by Ministerio de Economía y Competi-tividad (Spain) grants BFU2009-08967 and BFU2012-37146 and Regional Government Junta de Castilla y León (Spain) grants BIO103/VA45/11, VA145U13, and BIO/VA33/13. María Calvo was supported by a predoctoral fellowship from Regional Government Junta de Castilla y León (Spain) and the European Social Fund.
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Calvo, M., Villalobos, C., Núñez, L. (2015). Calcium Imaging in Neuron Cell Death. In: Lossi, L., Merighi, A. (eds) Neuronal Cell Death. Methods in Molecular Biology, vol 1254. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2152-2_6
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DOI: https://doi.org/10.1007/978-1-4939-2152-2_6
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