Abstract
Connexin proteins are the building blocks of gap junctions and connexin hemichannels. Both provide a pathway for cellular communication. Gap junctions support intercellular communication mechanisms and regulate homeostasis. In contrast, open connexin hemichannels connect the intracellular compartment and the extracellular environment, and their activation fuels inflammation and cell death. The development of clinically applicable connexin hemichannel blockers for therapeutic purposes is therefore gaining momentum. This chapter describes a well-established protocol optimized for assessing connexin hemichannel activity by using the reporter dye Yo-Pro1. A schematic depicts the presence of magnesium and calcium ions in the media when the C x 43 hemichannel is closed, whereas the media lacks these ions when the channel opens. Y o-P r o 1 dye permeates through the open hemichannel and allows the binding of C x 43 in the nucleus. A schematic depicts the presence of magnesium and calcium ions in the media when the C x 43 hemichannel is closed, whereas the media lacks these ions when the channel opens. Y o-P r o 1 dye permeates through the open hemichannel and allows the binding of C x 43 in the nucleus. A schematic depicts the presence of magnesium and calcium ions in the media when the C x 43 hemichannel is closed, whereas the media lacks these ions when the channel opens. Y o-P r o 1 dye permeates through the open hemichannel and allows the binding of C x 43 in the nucleus.
Original language | English |
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Article number | 6 |
Pages (from-to) | 75-85 |
Number of pages | 11 |
Journal | Methods in Molecular Biology |
Volume | 2801 |
DOIs | |
Publication status | Published - 6 Apr 2024 |
Bibliographical note
Funding Information:This work was financially supported by the European Research Council (Proof-of-Concept grant 861913), the European Future and Emerging Technologies program (grant 858014), the Research Foundation Flanders-Belgium (grants G012318N, G020018N and G0F7219N), the Methusalem program of the Flemish Government and the University Hospital of the Vrije Universiteit Brussel-Belgium (Scientific Fund Willy Gepts). The authors wish to thank Fien Haenen and Arne Loosen for their dedicated technical assistance.1. Lukowicz-Bedford R, Farnsworth DR, Miller AC (2021) Connexinplexity: the spatial and temporal expression of connexin genes during vertebrate organogenesis. bioRxiv preprint
Publisher Copyright:
© The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2024.