RyR1 and RyR3 isoforms provide distinct intracellular Ca2+ signals in HEK 293 cells

Daniela Rossi, Ilenia Simeoni, Marcella Micheli, Martin Bootman, Peter Lipp, Paul D. Allen, Vincenzo Sorrentino

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

Ryanodine receptors (RyRs) are expressed on the endoplasmic reticulum of many cells, where they form intracellular Ca2+-release channels that participate in the generation of intracellular Ca2+ signals. Here we report studies on the intracellular localisation and functional properties of transfected RyR1 or RyR3 channels in HEK 293 cells. Immunofluorescence studies indicated that both RyR1 and RyR3 did not form clusters but were homogeneously distributed throughout the endoplasmic reticulum. Ca2+ release experiments showed that transfected RyR1 and RyR3 channels responded to caffeine, although with different sensitivity, generating a global release of Ca2+ from the entire endoplasmic reticulum. However, video imaging and confocal microscopy analysis revealed that, in RyR3-expressing cells, local spontaneous Ca2+ release events were observed. No such spontaneous activity was observed in RyR1-expressing cells or in control cells. Interestingly, the spontaneous release events observed in RyR3-expressing cells were restricted to one or two regions of the endoplasmic reticulum, suggesting the formation of a further subcellular organisation of RyR3 in Ca2+ release units. These results demonstrate that different RyR isoforms can engage in the generation of distinct intracellular Ca2+ signals in HEK 293 cells.

Original languageEnglish (US)
Pages (from-to)2497-2504
Number of pages8
JournalJournal of Cell Science
Volume115
Issue number12
StatePublished - Jun 15 2002
Externally publishedYes

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Ryanodine Receptor Calcium Release Channel
HEK293 Cells
Protein Isoforms
Endoplasmic Reticulum
Video Microscopy
Caffeine
Confocal Microscopy
Fluorescent Antibody Technique

Keywords

  • Calcium release
  • Calcium signalling
  • Calcium stores
  • Endoplasmic sarcoplasmic reticulum
  • Ryanodine receptor channels

ASJC Scopus subject areas

  • Cell Biology

Cite this

Rossi, D., Simeoni, I., Micheli, M., Bootman, M., Lipp, P., Allen, P. D., & Sorrentino, V. (2002). RyR1 and RyR3 isoforms provide distinct intracellular Ca2+ signals in HEK 293 cells. Journal of Cell Science, 115(12), 2497-2504.

RyR1 and RyR3 isoforms provide distinct intracellular Ca2+ signals in HEK 293 cells. / Rossi, Daniela; Simeoni, Ilenia; Micheli, Marcella; Bootman, Martin; Lipp, Peter; Allen, Paul D.; Sorrentino, Vincenzo.

In: Journal of Cell Science, Vol. 115, No. 12, 15.06.2002, p. 2497-2504.

Research output: Contribution to journalArticle

Rossi, D, Simeoni, I, Micheli, M, Bootman, M, Lipp, P, Allen, PD & Sorrentino, V 2002, 'RyR1 and RyR3 isoforms provide distinct intracellular Ca2+ signals in HEK 293 cells', Journal of Cell Science, vol. 115, no. 12, pp. 2497-2504.
Rossi D, Simeoni I, Micheli M, Bootman M, Lipp P, Allen PD et al. RyR1 and RyR3 isoforms provide distinct intracellular Ca2+ signals in HEK 293 cells. Journal of Cell Science. 2002 Jun 15;115(12):2497-2504.
Rossi, Daniela ; Simeoni, Ilenia ; Micheli, Marcella ; Bootman, Martin ; Lipp, Peter ; Allen, Paul D. ; Sorrentino, Vincenzo. / RyR1 and RyR3 isoforms provide distinct intracellular Ca2+ signals in HEK 293 cells. In: Journal of Cell Science. 2002 ; Vol. 115, No. 12. pp. 2497-2504.
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