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. 2024:2801:97-109.
doi: 10.1007/978-1-0716-3842-2_8.

Measurement of Ca2+ Uptake Through Connexin Hemichannels

Chiara Nardin  1   2 , Fabio Mammano  3   4
Affiliations

Measurement of Ca2+ Uptake Through Connexin Hemichannels

Chiara Nardin et al. Methods Mol Biol. 2024.

Abstract

Increasing evidence points to deregulated flux of ionized calcium (Ca2+) mediated by hyperactive mutant connexin (Cx) hemichannels (HCs) as a common gain-of-function etiopathogenetic mechanism for several diseases, ranging from skin disorders to nervous system defects. Furthermore, the opening of nonmutated Cx HCs is associated with an impressive list of widespread diseases including, but not limited to, ischemia/stroke, Alzheimer's disease, and epilepsy. HC inhibitors are attracting a growing attention due to their therapeutic potential for numerous pathologies. This chapter describes a quantitative method to measure Ca2+ uptake though HCs expressed in cultured cells. The assay we developed can be used to probe HC activity as wells as to test HC inhibitors. Furthermore, with minor changes it can be easily adapted to high-throughput high-content platforms and/or primary cells and microtissues.

Keywords: Connexin43; GCaMP6s; HaCaT cells; Hemichannel functionality; Ionomycin; Lentiviral transduction; TAT-Gap19.

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References

    1. Maeda S, Nakagawa S, Suga M et al (2009) Structure of the connexin 26 gap junction channel at 3.5 A resolution. Nature 458(7238):597–602. https://doi.org/10.1038/nature07869 - DOI - PubMed
    1. Myers JB, Haddad BG, O’Neill SE et al (2018) Structure of native lens connexin 46/50 intercellular channels by cryo-EM. Nature 564(7736):372–377. https://doi.org/10.1038/s41586-018-0786-7 - DOI - PubMed - PMC
    1. Lee HJ, Jeong H, Hyun J et al (2020) Cryo-EM structure of human Cx31.3/GJC3 connexin hemichannel. Sci Adv 6(35):eaba4996. https://doi.org/10.1126/sciadv.aba4996 - DOI - PubMed - PMC
    1. Khan AK, Jagielnicki M, Bennett BC et al (2021) Cryo-EM structure of an open conformation of a gap junction hemichannel in lipid bilayer nanodiscs. Structure. https://doi.org/10.1016/j.str.2021.05.010
    1. Wang N, De Bock M, Decrock E et al (2013) Paracrine signaling through plasma membrane hemichannels. Biochim Biophys Acta 1828(1):35–50. https://doi.org/10.1016/j.bbamem.2012.07.002 - DOI - PubMed

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