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Cyclic ADP ribose is a novel regulator of intracellular Ca2+ oscillations in human bone marrow mesenchymal stem cells.

Tao R, Sun HY, Lau CP, Tse HF, Lee HC, Li GR - J. Cell. Mol. Med. (2011)

Bottom Line: However, cADPR had no effect on adipogenesis or osteogenesis in human MSCs.Our results indicate that cADPR is a novel regulator of Ca(2+) (i) oscillations in human MSCs.It permeates the cell membrane through the nucleoside transporters and increases Ca(2+) oscillation via activation of the TRPM2 channel, resulting in enhanced phosphorylation of ERK1/2 and, thereby, stimulation of human MSC proliferation.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.

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TRPM2 channel and cADPR in human MSCs. (A) Cyclic ADP ribose increased frequency of Ca2+i oscillations in a representative cell. (B) Cyclic ADP ribose had no significant effect on Ca2+i oscillations in the cell pre-treated with the TRMP2 channel inhibitor clotrimazole. (C) Econazole prevented cADPR effect on Ca2+i oscillations. (D) Mean values for the effects of 50 μM cADPR (n = 36), 10 μM clotrimazole plus 50 μM cADPR (n = 23) and 10 μM econazole plus 50 μM cADPR (n = 26) on frequency of Ca2+i oscillations. *P < 0.05 versus before cADPR. (E) RT-PCR revealed the expression of full length isoform of TRPM2 (TRPM2-L) gene. (F) Western blot analysis showed TRPM2 channel protein in human MSCs from passages 2, 5 and 8.
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fig04: TRPM2 channel and cADPR in human MSCs. (A) Cyclic ADP ribose increased frequency of Ca2+i oscillations in a representative cell. (B) Cyclic ADP ribose had no significant effect on Ca2+i oscillations in the cell pre-treated with the TRMP2 channel inhibitor clotrimazole. (C) Econazole prevented cADPR effect on Ca2+i oscillations. (D) Mean values for the effects of 50 μM cADPR (n = 36), 10 μM clotrimazole plus 50 μM cADPR (n = 23) and 10 μM econazole plus 50 μM cADPR (n = 26) on frequency of Ca2+i oscillations. *P < 0.05 versus before cADPR. (E) RT-PCR revealed the expression of full length isoform of TRPM2 (TRPM2-L) gene. (F) Western blot analysis showed TRPM2 channel protein in human MSCs from passages 2, 5 and 8.

Mentions: To investigate whether TRPM2 channels participate in the cADPR-induced increase of Ca2+i oscillation frequency, two structurally different inhibitors of the channel, econazole (10 μM) and clotrimazole (10 μM) [41], were tested in human MSCs. Both showed no significant effect on the spontaneous Ca2+i oscillations, but both effectively prevented the increase of Ca2+i oscillation frequency induced by cADPR (Fig. 4B and C). The quantitative results are summarized in Figure 4D. RT-PCR (Fig. 4E) showed the presence of the mRNA for the full length form of TRPM2, TRPM2-L, but not short form, TRPM2-S, which was also confirmed by Western blot analysis (Fig. 4F) for protein expression in human MSCs. These results suggest that the increased Ca2+i oscillation frequency by cADPR is most likely mediated by the TRPM2 channels.


Cyclic ADP ribose is a novel regulator of intracellular Ca2+ oscillations in human bone marrow mesenchymal stem cells.

Tao R, Sun HY, Lau CP, Tse HF, Lee HC, Li GR - J. Cell. Mol. Med. (2011)

TRPM2 channel and cADPR in human MSCs. (A) Cyclic ADP ribose increased frequency of Ca2+i oscillations in a representative cell. (B) Cyclic ADP ribose had no significant effect on Ca2+i oscillations in the cell pre-treated with the TRMP2 channel inhibitor clotrimazole. (C) Econazole prevented cADPR effect on Ca2+i oscillations. (D) Mean values for the effects of 50 μM cADPR (n = 36), 10 μM clotrimazole plus 50 μM cADPR (n = 23) and 10 μM econazole plus 50 μM cADPR (n = 26) on frequency of Ca2+i oscillations. *P < 0.05 versus before cADPR. (E) RT-PCR revealed the expression of full length isoform of TRPM2 (TRPM2-L) gene. (F) Western blot analysis showed TRPM2 channel protein in human MSCs from passages 2, 5 and 8.
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fig04: TRPM2 channel and cADPR in human MSCs. (A) Cyclic ADP ribose increased frequency of Ca2+i oscillations in a representative cell. (B) Cyclic ADP ribose had no significant effect on Ca2+i oscillations in the cell pre-treated with the TRMP2 channel inhibitor clotrimazole. (C) Econazole prevented cADPR effect on Ca2+i oscillations. (D) Mean values for the effects of 50 μM cADPR (n = 36), 10 μM clotrimazole plus 50 μM cADPR (n = 23) and 10 μM econazole plus 50 μM cADPR (n = 26) on frequency of Ca2+i oscillations. *P < 0.05 versus before cADPR. (E) RT-PCR revealed the expression of full length isoform of TRPM2 (TRPM2-L) gene. (F) Western blot analysis showed TRPM2 channel protein in human MSCs from passages 2, 5 and 8.
Mentions: To investigate whether TRPM2 channels participate in the cADPR-induced increase of Ca2+i oscillation frequency, two structurally different inhibitors of the channel, econazole (10 μM) and clotrimazole (10 μM) [41], were tested in human MSCs. Both showed no significant effect on the spontaneous Ca2+i oscillations, but both effectively prevented the increase of Ca2+i oscillation frequency induced by cADPR (Fig. 4B and C). The quantitative results are summarized in Figure 4D. RT-PCR (Fig. 4E) showed the presence of the mRNA for the full length form of TRPM2, TRPM2-L, but not short form, TRPM2-S, which was also confirmed by Western blot analysis (Fig. 4F) for protein expression in human MSCs. These results suggest that the increased Ca2+i oscillation frequency by cADPR is most likely mediated by the TRPM2 channels.

Bottom Line: However, cADPR had no effect on adipogenesis or osteogenesis in human MSCs.Our results indicate that cADPR is a novel regulator of Ca(2+) (i) oscillations in human MSCs.It permeates the cell membrane through the nucleoside transporters and increases Ca(2+) oscillation via activation of the TRPM2 channel, resulting in enhanced phosphorylation of ERK1/2 and, thereby, stimulation of human MSC proliferation.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.

Show MeSH