Measurement of Ca 2+ Uptake Through Connexin Hemichannels

Increasing evidence points to deregulated flux of ionized calcium (Ca ) 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 non...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 2801; p. 97
Main Authors Nardin, Chiara, Mammano, Fabio
Format Journal Article
LanguageEnglish
Published United States 2024
Subjects
Biological Transport
Calcium - metabolism
Connexin 43 - metabolism
Connexins - genetics
Connexins - metabolism
Ionomycin
HaCaT cells
TAT-Gap19
Hemichannel functionality
Lentiviral transduction
Connexin43
GCaMP6s
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Summary:Increasing evidence points to deregulated flux of ionized calcium (Ca ) 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 Ca 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.
ISSN:1940-6029