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Indicator-dependent...
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Yang, MingyuUppsala universitet,Institutionen för medicinsk cellbiologi
(författare)
Indicator-dependent differences in detection of local intracellular Ca2+release events evoked by voltage-gated Ca2+entry in pancreatic & beta;-cells
- Artikel/kapitelEngelska2023
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Elsevier BV,2023
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electronicrdacarrier
Nummerbeteckningar
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LIBRIS-ID:oai:DiVA.org:uu-509244
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https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-509244URI
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https://doi.org/10.1016/j.cellsig.2023.110805DOI
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Språk:engelska
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Sammanfattning på:engelska
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Ämneskategori:ref swepub-contenttype
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Ämneskategori:art swepub-publicationtype
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Genetically encoded Ca2+ indicators have become widely used in cell signalling studies as they offer advantages over cell-loaded dye indicators in enabling specific cellular or subcellular targeting. Comparing responses from dye and protein-based indicators may provide information about indicator properties and cell physiology, but side-by-side recordings in cells are scarce. In this study, we compared cytoplasmic Ca2+ concentration ([Ca2+]i) changes in insulin-secreting & beta;-cells recorded with commonly used dyes and indicators based on circularly permuted fluorescent proteins. Total internal reflection fluorescence (TIRF) imaging of K+ depolarizationtriggered submembrane [Ca2+]i increases showed that the dyes Fluo-4 and Fluo-5F mainly reported stable [Ca2+]i elevations, whereas the proteins R-GECO1 and GCaMP5G more often reported distinct [Ca2+]i spikes from an elevated level. [Ca2+]i spiking occurred also in glucose-stimulated cells. The spikes reflected Ca2+ release from the endoplasmic reticulum, triggered by autocrine activation of purinergic receptors after exocytotic release of ATP and/or ADP, and the spikes were consequently prevented by SERCA inhibition or P2Y1-receptor antagonism. Widefield imaging, which monitors the entire cytoplasm, increased the spike detection by the Ca2+ dyes. The indicator-dependent response patterns were unrelated to Ca2+ binding affinity, buffering and mobility, and probably reflects the much slower dissociation kinetics of protein compared to dye indicators. Ca2+ dyes thus report signalling within the submembrane space excited by TIRF illumination, whereas the protein indicators also catch Ca2+ events originating outside this volume. The study highlights that voltage-dependent Ca2+ entry in & beta;-cells is tightly linked to local intracellular Ca2+ release mediated via an autocrine route that may be more important than previously reported direct Ca2+ effects on phospholipase C or on intracellular channels mediating calcium-induced calcium release.
Ämnesord och genrebeteckningar
Biuppslag (personer, institutioner, konferenser, titlar ...)
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Dyachok, Oleg,1965-Uppsala universitet,Institutionen för medicinsk cellbiologi(Swepub:uu)olegdyac
(författare)
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Xu, YunjianUppsala universitet,Institutionen för medicinsk cellbiologi(Swepub:uu)yunxu459
(författare)
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Gylfe, Erik,1947-Uppsala universitet,Institutionen för medicinsk cellbiologi(Swepub:uu)erikgylf
(författare)
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Idevall Hagren, Olof,1980-Uppsala universitet,Institutionen för medicinsk cellbiologi(Swepub:uu)olide169
(författare)
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Tengholm, Anders,1971-Uppsala universitet,Institutionen för medicinsk cellbiologi(Swepub:uu)andeteng
(författare)
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Uppsala universitetInstitutionen för medicinsk cellbiologi
(creator_code:org_t)
Sammanhörande titlar
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Ingår i:Cellular Signalling: Elsevier BV1090898-65681873-3913
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