title: TRPM2 channel membrane currents in primary rat megakaryocytes were activated by the agonist ADP-ribose but not oxidative stress.
abstract: Melastatin-like transient receptor potential 2 (TRPM2) channel activation/inhibition mechanisms in response to ADP-ribose (ADPR), oxidative stress, flufenamic acid (FFA) and 2-aminoethoxydiphenyl borate (2-APB) are not clear. We tested the effects of FFA and 2-APB on ADPR-induced TRPM2 cation channel currents in rat native bone marrow megakaryocytes. Megakaryocyte cells were freshly isolated from rat bone marrow and studied with the conventional whole-cell patch-clamp technique. Extracellular H2O2, FFA and 2-APB were added through the patch chamber, while intracellular ADPR was applied through the pipette. Nonselective cation currents were consistently induced by ADPR but not H2O2. Current density of ADPR in the cells was significantly (P<0.001) higher than in control. The time courses of ADPR effects in the megakaryocytes were characterized by a delay of 2.24±0.73. The ADPR-induced Ca2+ gate was not blocked by either the IP3 receptor inhibitor 2-APB or the PLC inhibitor FFA. In conclusion, TRPM2 channels were constitutively activated by intracellular ADPR, although the channel currents in rat native megakaryocytes were not affected by extracellular H2O2, 2-APB or FFA. Activation of TRPM2 channels in megakaryocytes seems to be intracellular and ADPR-dependent.
authors.affiliation: Department of Biophysics, Medical Faculty, Süleyman Demirel University, Morfoloji Binasi, 32260, Isparta, Turkey. mnaziroglu@med.sdu.edu.tr
authors.forename: Mustafa
authors.initials: M
authors.lastname: Nazıroğlu
mesh_terms: D000246:Adenosine Diphosphate Ribose / Q000494:pharmacology*; D000818:Animals; D001896:Boron Compounds / Q000494:pharmacology; D020013:Calcium Signaling; D005439:Flufenamic Acid / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000494:pharmacology; D066298:In Vitro Techniques; D015640:Ion Channel Gating / Q000187:drug effects; D008297:Male; D008533:Megakaryocytes / Q000187:drug effects / Q000502:physiology*; D008564:Membrane Potentials / Q000187:drug effects*; D018384:Oxidative Stress; D018408:Patch-Clamp Techniques; D051381:Rats; D017208:Rats, Wistar; D050053:TRPM Cation Channels / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D001896:Boron Compounds; D050053:TRPM Cation Channels; C557080:Trpm2 protein, rat; D000246:Adenosine Diphosphate Ribose; D005439:Flufenamic Acid; D006861:Hydrogen Peroxide; C109986:2-aminoethoxydiphenyl borate