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Measuring Connexin Hemichannel Opening in Response to an InsP3-Mediated Cytosolic Ca2+ Increase

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Connexin Hemichannels

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2801))

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Abstract

The opening of connexin hemichannels (HCs) expressed at the plasma membrane of mammalian cells is regulated by a number of physiological parameters, including extracellular and intracellular Ca2+ ions. Submicromolar variations of the cytosolic Ca2+ concentration ([Ca2+]c) are per se sufficient to trigger extracellular bursts of messenger molecules through connexin HCs, thus mediating paracrine signaling. In this chapter, we present a quantitative method to measure the opening dynamics of connexin HCs expressed in a single HeLa cell upon stimulation by a canonical InsP3-mediated [Ca2+]c transient. The protocol relies on a combination of Ca2+ imaging and patch-clamp techniques. The insights gained from our method are expected to make a significant contribution to understanding the structure–function relationship of connexin HCs. The protocol is also suitable to screen candidate therapeutic compounds to treat connexin-related diseases linked to HC dysfunction.

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Acknowledgments

This work was funded by grants awarded to Mario Bortolozzi by the French Telethon Foundation and by Theraphies for Inherited Neuropathies (TIN US) non-profit organization.

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Authors and Affiliations

  1. Department of Physics and Astronomy “G. Galilei”, University of Padua, Padova, Italy

    Erva Bayraktar & Mario Bortolozzi

  2. Veneto Institute of Molecular Medicine (VIMM), Padova, Italy

    Erva Bayraktar & Mario Bortolozzi

Authors
  1. Erva Bayraktar
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  2. Mario Bortolozzi
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Corresponding author

Correspondence to Mario Bortolozzi .

Editor information

Editors and Affiliations

  1. Department of Physics, University of Padova, Padova, Italy

    Fabio Mammano

  2. Facultad de Medicina, Universidad del Desarrollo, Santiago, Chile

    Mauricio Retamal

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Bayraktar, E., Bortolozzi, M. (2024). Measuring Connexin Hemichannel Opening in Response to an InsP3-Mediated Cytosolic Ca2+ Increase. In: Mammano, F., Retamal, M. (eds) Connexin Hemichannels. Methods in Molecular Biology, vol 2801. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3842-2_14

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  • DOI: https://doi.org/10.1007/978-1-0716-3842-2_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3841-5

  • Online ISBN: 978-1-0716-3842-2

  • eBook Packages: Springer Protocols

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