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Both RyRs and TPCs are required for NAADP-induced intracellular Ca²⁺ release.

Gerasimenko JV, Charlesworth RM, Sherwood MW, Ferdek PE, Mikoshiba K, Parrington J, Petersen OH, Gerasimenko OV - Cell Calcium (2015)

Bottom Line: Antibodies against RyR2 had practically no effect on NAADP-evoked Ca(2+) release, but reduced release in response to cADPR by 55%.We conclude that full NAADP-mediated Ca(2+) release requires both TPCs and RyRs.Our results indicate that the primary, but very small, NAADP-elicited Ca(2+) release via TPCs from endosomes/lysosomes triggers the detectable Ca(2+)-induced Ca(2+) release via RyR1 and RyR3 occurring from the granules and the ER.

View Article: PubMed Central - PubMed

Affiliation: Medical Research Council Group, Cardiff School of Biosciences, Cardiff University, Cardiff, Wales, UK.

No MeSH data available.


Related in: MedlinePlus

Inhibitory effect of Ned-19 (100 μM) on NAADP-induced signalling in permeabilized cells. (A) Representative trace of application of 100 nM NAADP to permeabilized pancreatic acinar cells. (B) Representative trace of application of 100 nM NAADP followed by 10 μM IP3 to permeabilized pancreatic acinar cells in the presence of 100 μM Ned-19. (C) Representative trace of application of 10 μM cADPR to permeabilized pancreatic acinar cells. (D) Representative trace of application of 10 μM cADPR followed by 10 μM IP3 to permeabilized pancreatic acinar cells in the presence of 100 μM Ned-19. (E) Summary of data comparison of applications of NAADP (control 26.7 ± 1.8%, n = 13; with Ned-19 8.6 ± 1.1, n = 5, p = 0.00002), cADPR (control 21.2 ± 0.9%, n = 10; with Ned-19 18.3 ± 0.9%, n = 4; p = 0.1) or IP3 (control 23.6 ± 1.5%, n = 11; with Ned-19 21.2 ± 1.1%, n = 6, p = 0.3) in the presence and absence of Ned-19. Error bars represent ± SEM. Cells were loaded with Fluo-5N in AM form.
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fig0010: Inhibitory effect of Ned-19 (100 μM) on NAADP-induced signalling in permeabilized cells. (A) Representative trace of application of 100 nM NAADP to permeabilized pancreatic acinar cells. (B) Representative trace of application of 100 nM NAADP followed by 10 μM IP3 to permeabilized pancreatic acinar cells in the presence of 100 μM Ned-19. (C) Representative trace of application of 10 μM cADPR to permeabilized pancreatic acinar cells. (D) Representative trace of application of 10 μM cADPR followed by 10 μM IP3 to permeabilized pancreatic acinar cells in the presence of 100 μM Ned-19. (E) Summary of data comparison of applications of NAADP (control 26.7 ± 1.8%, n = 13; with Ned-19 8.6 ± 1.1, n = 5, p = 0.00002), cADPR (control 21.2 ± 0.9%, n = 10; with Ned-19 18.3 ± 0.9%, n = 4; p = 0.1) or IP3 (control 23.6 ± 1.5%, n = 11; with Ned-19 21.2 ± 1.1%, n = 6, p = 0.3) in the presence and absence of Ned-19. Error bars represent ± SEM. Cells were loaded with Fluo-5N in AM form.

Mentions: In two-photon permeabilized pancreatic acinar cells, pre-incubation with a high concentration of Ned-19 (100 μM) inhibited NAADP-elicited Ca2+ responses by 67.8% (Fig. 2A, B, and E), whereas responses to cADPR (10 μM) (Fig. 2C–E) and IP3 (Fig. 2E) were not significantly affected, indicating that the action of Ned-19 is specific for NAADP-induced responses.


Both RyRs and TPCs are required for NAADP-induced intracellular Ca²⁺ release.

Gerasimenko JV, Charlesworth RM, Sherwood MW, Ferdek PE, Mikoshiba K, Parrington J, Petersen OH, Gerasimenko OV - Cell Calcium (2015)

Inhibitory effect of Ned-19 (100 μM) on NAADP-induced signalling in permeabilized cells. (A) Representative trace of application of 100 nM NAADP to permeabilized pancreatic acinar cells. (B) Representative trace of application of 100 nM NAADP followed by 10 μM IP3 to permeabilized pancreatic acinar cells in the presence of 100 μM Ned-19. (C) Representative trace of application of 10 μM cADPR to permeabilized pancreatic acinar cells. (D) Representative trace of application of 10 μM cADPR followed by 10 μM IP3 to permeabilized pancreatic acinar cells in the presence of 100 μM Ned-19. (E) Summary of data comparison of applications of NAADP (control 26.7 ± 1.8%, n = 13; with Ned-19 8.6 ± 1.1, n = 5, p = 0.00002), cADPR (control 21.2 ± 0.9%, n = 10; with Ned-19 18.3 ± 0.9%, n = 4; p = 0.1) or IP3 (control 23.6 ± 1.5%, n = 11; with Ned-19 21.2 ± 1.1%, n = 6, p = 0.3) in the presence and absence of Ned-19. Error bars represent ± SEM. Cells were loaded with Fluo-5N in AM form.
© Copyright Policy - CC BY
Related In: Results  -  Collection

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fig0010: Inhibitory effect of Ned-19 (100 μM) on NAADP-induced signalling in permeabilized cells. (A) Representative trace of application of 100 nM NAADP to permeabilized pancreatic acinar cells. (B) Representative trace of application of 100 nM NAADP followed by 10 μM IP3 to permeabilized pancreatic acinar cells in the presence of 100 μM Ned-19. (C) Representative trace of application of 10 μM cADPR to permeabilized pancreatic acinar cells. (D) Representative trace of application of 10 μM cADPR followed by 10 μM IP3 to permeabilized pancreatic acinar cells in the presence of 100 μM Ned-19. (E) Summary of data comparison of applications of NAADP (control 26.7 ± 1.8%, n = 13; with Ned-19 8.6 ± 1.1, n = 5, p = 0.00002), cADPR (control 21.2 ± 0.9%, n = 10; with Ned-19 18.3 ± 0.9%, n = 4; p = 0.1) or IP3 (control 23.6 ± 1.5%, n = 11; with Ned-19 21.2 ± 1.1%, n = 6, p = 0.3) in the presence and absence of Ned-19. Error bars represent ± SEM. Cells were loaded with Fluo-5N in AM form.
Mentions: In two-photon permeabilized pancreatic acinar cells, pre-incubation with a high concentration of Ned-19 (100 μM) inhibited NAADP-elicited Ca2+ responses by 67.8% (Fig. 2A, B, and E), whereas responses to cADPR (10 μM) (Fig. 2C–E) and IP3 (Fig. 2E) were not significantly affected, indicating that the action of Ned-19 is specific for NAADP-induced responses.

Bottom Line: Antibodies against RyR2 had practically no effect on NAADP-evoked Ca(2+) release, but reduced release in response to cADPR by 55%.We conclude that full NAADP-mediated Ca(2+) release requires both TPCs and RyRs.Our results indicate that the primary, but very small, NAADP-elicited Ca(2+) release via TPCs from endosomes/lysosomes triggers the detectable Ca(2+)-induced Ca(2+) release via RyR1 and RyR3 occurring from the granules and the ER.

View Article: PubMed Central - PubMed

Affiliation: Medical Research Council Group, Cardiff School of Biosciences, Cardiff University, Cardiff, Wales, UK.

No MeSH data available.


Related in: MedlinePlus