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Intracellular Calcium Measurements for Functional Characterization of Neuronal Phenotypes

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Embryonic Stem Cell Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1341))

Abstract

The central and peripheral nervous system is built by a network of many different neuronal phenotypes together with glial and other supporting cells. The repertoire of expressed receptors and secreted neurotransmitters and neuromodulators are unique for each single neuron leading to intracellular signaling cascades, many of them involving intracellular calcium signaling. Here we suggest the use of calcium signaling analysis upon specific agonist application to reliably identify neuronal phenotypes, being important not only for basic science, but also providing a reliable tool for functional characterization of cells prior to transplantation. Calcium imaging provides various cellular information including signaling amplitudes, cell localization, duration, and frequency. Microfluorimetry reveals a signal summarizing the entire population, and its use is indicated for high-throughput screening purposes.

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References

  1. Gaspard N, Vanderhaeghen P (2010) Mechanisms of neural specification from embryonic stem cells. Curr Opin Neurobiol 20:37–43

    Article  CAS  PubMed  Google Scholar 

  2. Smith AG (2001) Embryo-derived stem cells: of mice and men. Annu Rev Cell Dev Biol 17:435–462

    Article  CAS  PubMed  Google Scholar 

  3. Calloni R, Cordero EA, Henriques JA et al (2013) Reviewing and updating the major molecular markers for stem cells. Stem Cells Dev 22:1455–1476

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  4. Berridge MJ, Lipp P, Bootman MD (2000) The versatility and universality of calcium signalling. Nat Rev Mol Cell Biol 1:11–21

    Article  CAS  PubMed  Google Scholar 

  5. Vanaman T, Carafoli E (2012) Introduction to thematic minireview series on calcium. J Biol Chem 287:31623

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  6. Parekh AB (2011) Decoding cytosolic Ca2+ oscillations. Trends Biochem Sci 36:78–87

    Article  CAS  PubMed  Google Scholar 

  7. Cheng H, Lederer WJ (2008) Calcium sparks. Physiol Rev 88:1491–1545

    Article  CAS  PubMed  Google Scholar 

  8. Berridge MJ (2006) Calcium microdomains: organization and function. Cell Calcium 40:405–412

    Article  CAS  PubMed  Google Scholar 

  9. Buonanno A, Fields RD (1999) Gene regulation by patterned electrical activity during neural and skeletal muscle development. Curr Opin Neurobiol 9:110–120

    Article  CAS  PubMed  Google Scholar 

  10. Lin JH, Takano T, Arcuino G et al (2007) Purinergic signaling regulates neural progenitor cell expansion and neurogenesis. Dev Biol 302:356–366

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  11. Spitzer NC, Lautermilch NJ, Smith RD et al (2000) Coding of neuronal differentiation by calcium transients. Bioessays 22:811–817

    Article  CAS  PubMed  Google Scholar 

  12. Gu X, Spitzer NC (1995) Distinct aspects of neuronal differentiation encoded by frequency of spontaneous Ca2+ transients. Nature 375:784–787

    Article  CAS  PubMed  Google Scholar 

  13. Simpson PB, Challiss RA, Nahorski SR (1995) Neuronal Ca2+ stores: activation and function. Trends Neurosci 18:299–306

    Article  CAS  PubMed  Google Scholar 

  14. Strubing C, Ahnert-Hilger G, Shan J et al (1995) Differentiation of pluripotent embryonic stem cells into the neuronal lineage in vitro gives rise to mature inhibitory and excitatory neurons. Mech Dev 53:275–287

    Article  CAS  PubMed  Google Scholar 

  15. Grienberger C, Konnerth A (2012) Imaging calcium in neurons. Neuron 73:862–885

    Article  CAS  PubMed  Google Scholar 

  16. Abranches E, Silva M, Pradier L et al (2009) Neural differentiation of embryonic stem cells in vitro: a road map to neurogenesis in the embryo. PLoS One 4:e6286

    Article  PubMed Central  PubMed  Google Scholar 

  17. Salti A, Nat R, Neto S et al (2013) Expression of early developmental markers predicts the efficiency of embryonic stem cell differentiation into midbrain dopaminergic neurons. Stem Cells Dev 22:397–411

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  18. Bibel M, Richter J, Lacroix E et al (2007) Generation of a defined and uniform population of CNS progenitors and neurons from mouse embryonic stem cells. Nat Protoc 2:1034–1043

    Article  CAS  PubMed  Google Scholar 

  19. Wichterle H, Lieberam I, Porter JA et al (2002) Directed differentiation of embryonic stem cells into motor neurons. Cell 110:385–397

    Article  CAS  PubMed  Google Scholar 

  20. Lau T, Schneidt T, Heimann F et al (2010) Somatodendritic serotonin release and re-uptake in mouse embryonic stem cell-derived serotonergic neurons. Neurochem Int 57:969–978

    Article  CAS  PubMed  Google Scholar 

  21. Maisano X, Litvina E, Tagliatela S et al (2012) Differentiation and functional incorporation of embryonic stem cell-derived GABAergic interneurons in the dentate gyrus of mice with temporal lobe epilepsy. J Neurosci 32:46–61

    Article  PubMed Central  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was supported by research grants from Brazilian funding agencies Sao Paulo Research Foundation (FAPESP), National Council for Scientific and Technological Development (CNPq), and Provost’s Office for Research of the University of São Paulo, Grant number: 2011.1.9333.1.3 (NAPNA-USP), Brazil.

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Authors and Affiliations

  1. Department of Biochemistry, Institute of Chemistry, University of São Paulo, Av. Prof. Lineu Prestes 748, São Paulo, Brazil

    Talita Glaser, Ana Regina G. Castillo, Ágatha Oliveira & Henning Ulrich

Authors
  1. Talita Glaser
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  2. Ana Regina G. Castillo
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  3. Ágatha Oliveira
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  4. Henning Ulrich
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Corresponding author

Correspondence to Henning Ulrich .

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Editors and Affiliations

  1. Ottawa Hospital Research Institute, Ottawa, Ontario, Canada

    Kursad Turksen

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© 2015 Springer Science+Business Media New York

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Glaser, T., Castillo, A.R.G., Oliveira, Á., Ulrich, H. (2015). Intracellular Calcium Measurements for Functional Characterization of Neuronal Phenotypes. In: Turksen, K. (eds) Embryonic Stem Cell Protocols. Methods in Molecular Biology, vol 1341. Humana Press, New York, NY. https://doi.org/10.1007/7651_2015_271

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  • DOI: https://doi.org/10.1007/7651_2015_271

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2953-5

  • Online ISBN: 978-1-4939-2954-2

  • eBook Packages: Springer Protocols

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