Book chapter
Techniques for Simultaneous Mitochondrial and Cytosolic Ca2+ Imaging in Neurons
Techniques to Investigate Mitochondrial Function in Neurons, pp.151-178
Neuromethods, Springer New York
03/18/2017
DOI: 10.1007/978-1-4939-6890-9_8
Abstract
Mitochondria efficiently buffer Ca2+ influx during excitation, which limits the amplitude of cytosolic Ca2+ rise, and then slowly release Ca2+ back into the cytosol thereby extending the duration of cytosolic Ca2+ response. This mitochondrial Ca2+ cycling helps shape Ca2+ transients and regulates Ca2+-dependent functions in neurons such as excitability, synaptic plasticity, bioenergetics, and survival. Therefore identifying the molecular components of mitochondrial Ca2+ transport in neurons and defining their pharmacological and functional properties presents an important and challenging task for neuroscientists. Fulfilling this task requires a set of tools for simple and reliable measurement of Ca2+ concentration inside and outside mitochondria, i.e., within the mitochondrial matrix and cytosol, respectively. In this chapter we describe instrumentation and techniques for simultaneous measurements of mitoc hondrial and cytosolic Ca2+ concentration in central and peripheral neurons by using synthetic (e.g., Fura-2) and genetic (e.g., mito-R-GECO1) fluorescent Ca2+ indicators. We include detailed protocols for preparing primary cultures of hippocampal and dorsal root ganglion (DRG) sensory neurons, gene transfer into these cells and the use of fluorescent microscopy and patch clampatch clampp for comprehensive characterization of Ca2+ fluxes across the mitochondrial and plasma membranes.
Details
- Title: Subtitle
- Techniques for Simultaneous Mitochondrial and Cytosolic Ca2+ Imaging in Neurons
- Creators
- Jacob E Rysted - Department of Pharmacology, University of Iowa Carver College of Medicine, Iowa City, USAZhihong Lin - Department of Pharmacology, University of Iowa Carver College of Medicine, Iowa City, USAYuriy M Usachev - Department of Pharmacology, University of Iowa Carver College of Medicine, Iowa City, USA
- Resource Type
- Book chapter
- Publication Details
- Techniques to Investigate Mitochondrial Function in Neurons, pp.151-178
- Publisher
- Springer New York; New York, NY
- Series
- Neuromethods
- DOI
- 10.1007/978-1-4939-6890-9_8
- eISSN
- 1940-6045
- ISSN
- 0893-2336
- Language
- English
- Date published
- 03/18/2017
- Academic Unit
- Neuroscience and Pharmacology; Anesthesia; Iowa Neuroscience Institute; Neurology; Fraternal Order of Eagles Diabetes Research Center
- Record Identifier
- 9984070496502771
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