Activation of membrane TRPM2 channel currents in primary Kv1.3-deficient mice megakaryocyte by agonist ADP-Ribose

Abstract

Since the mechanisms that lead to melastatin-like transient receptor potential 2 (TRPM2) channel activation in response to ADP-Ribose (ADPR) are not understood, I tested the effects of ADP-Ribose on TRPM2 cation channel currents in native bone marrow megakaryocyte of mice. Kv1.3-deficient mice megakaryocyte cells were freshly isolated from bone marrow and the cells studied with the conventional whole-cell patch clamp technique. ADPR (1mM) was applied intracellularly through the pipette. Non-selective cation currents in the mice were consistently induced by ADPR. Current density of ADPR in the cells were significantly (p<0.001) higher than in control. The time courses of ADPR effects in the mice were characterized by a delay of 3.66 ± 0.69. In conclusion, TRPM2 channels were constitutively activated by intracellular ADPR currents in mice native megakaryocyte.

Keywords

• TRPM2 channels
• ADP-Ribose
• bone marrow
• megakaryocyte

References

1. Bai JZ, Lipski J. 2010. Differential expression of TRPM2 and TRPV4 channels and their potential role in oxidative stress-induced cell death in organotypic hippocampal culture. Neurotoxicology 31:204-214.
2. Buelow B, Song Y, Scharenberg AM. 2008. The Poly(ADP-ribose) polymerase PARP-1 is required for oxidative stress-induced TRPM2 activation in lymphocytes. J Biol Chem. 283:24571-24583.
3. Carter RN, Tolhurst G, Walmsley G, Vizuete-Forster M, Miller N, Mahaut-Smith MP. 2006. Molecular and electrophysiological characterization of transient receptor potential ion channels in the primary murine megakaryocyte. J Physiol. 576:151-162.
4. Guse AH. 2005. Second messenger function and the structure-activity relationship of cyclic adenosine diphosphoribose (cADPR). FEBS J 272: 4590-4597.
5. Hara Y, Wakamori M, Ishii M, Maeno E, Nishida M, Yoshida T, et al., 2002. LTRPC2 Ca2+-permeable channel activated by changes in redox status confers susceptibility to cell death. Mol Cell 9: 163-173.
6. Heiner I, Eisfeld J, Warnstedt M, Radukina N, Jungling E, Luckhoff A. 2006. Endogenous ADP-ribose enables calcium-regulated cation currents through TRPM2 channels in neutrophil granulocytes. Biochem J 398: 225- 232.
7. Ishii M, Shimizu S, Hara Y, Hagiwara T, Miyazaki A, Mori Y, Kiuchi Y. 2006. Intracellular-produced hydroxyl radical mediates H2O2-induced Ca2+ influx and cell death in rat beta-cell line RIN-5F. Cell Calcium 39:487-494.
8. Mahaut-Smith MP. 2004. Patch-clamp recordings of electrophysiological events in the platelet and megakaryocyte. Methods Mol Biol. 273:277-300.

9. Nazıroğlu M. 2007. New molecular mechanisms on the activation of TRPM2 channels by oxidative stress and ADP-ribose. Neurochem Res 32:1990- 2001.
10. Nazıroğlu M, Lückhoff A. 2008a. Effects of antioxidants on calcium influx through TRPM2 channels in transfected cells activated by hydrogen peroxide. J Neurol Sci 270:152-158.
11. Nazıroğlu M, Lückhoff A. 2008b. A calcium influx pathway regulated separately by oxidative stress and ADP-Ribose in TRPM2 channels: single channel events. Neurochem Res 33:1256-1262.
12. Nazıroğlu M. 2010. TRPM2 cation channels, oxidative stress and neurological diseases: where are we now? Neurochem Res DOI: 10.1007/s11064-010- 0347-4.
13. Perraud AL, Fleig A, Dunn CA, Bagley LA, Launay P, Schmitz C, Stokes AJ, Zhu Q, Bessman MJ, Penner R, Kinet JP, Scharenberg AM. 2001. ADP-ribose gating of the calcium-permeable LTRPC2 channel revealed by Nudix motif homology. Nature 411:595-599.
14. Perraud AL, Takanishi CL, Shen B, Kang S, Smith MK, Schmitz C, Knowles HM, Ferraris D, Li W, Zhang J, Stoddard BL, Scharenberg AM. 2005. Accumulation of free ADP-ribose from mitochondria mediates oxidative stress-induced gating of TRPM2 cation channels. J Biol Chem 280:6138- 6148.
15. Tolhurst G, Carter RN, Amisten S, Holdich JP, Erlinge D, Mahaut-Smith MP. 2008. Expression profiling and electrophysiological studies suggest a major role for Orai1 in the store-operated Ca2+ influx pathway of platelets and megakaryocytes. Platelets. 19:308-313.
16. Wehage E, Eisfeld J, Heiner I, Jüngling E, Zitt C, Lückhoff A. 2002. Activation of the cation channel long transient receptor potential channel 2 (LTRPC2) by hydrogen peroxide. A splice variant reveals a mode of activation independent of ADP-ribose. J Biol Chem 277: 23150-23156.
17. Wilkinson JA, Scragg JL, Boyle JP, Nilius B, Peers C. 2008. H2O2-stimulated Ca2+ influx via TRPM2 is not the sole determinant of subsequent cell death. Pflugers Arch. 455:1141-1151.
18. Xu SZ, Zeng F, Boulay G, Grimm C, Harteneck C, Beech DJ. 2005. Block of TRPC5 channels by 2-aminoethoxydiphenyl borate: a differential, extracellular and voltage-dependent effect. Br J Pharmacol. 2005;145:405-414.