NDLI: A Calcium Influx Pathway Regulated Separately by Oxidative Stress and ADP-Ribose in TRPM2 Channels: Single Channel Events

Content Provider	Springer Nature Link
Author	Nazıroğlu, Mustafa Lückhoff, Andreas
Copyright Year	2008
Abstract	A melastatin-like transient receptor potential 2 (TRPM2) channel is activated in concert with Ca$^{2+}$ by intracellular adenosine diphosphoribose (ADPR) binding to the channel’s enzyme Nudix domain. Channel activity is also seen with nicotinamide dinucleotide (NAD$^{+}$) and hydrogen peroxide (H$_{2}$O$_{2}$) although the mechanisms remain unknown. Hence, we tested the effects of ADPR, NAD$^{+}$ and H$_{2}$O$_{2}$ on the activation of TRPM2 currents in transfected Chinese hamster ovary (CHO) cells. The CHO cells were transfected with cDNA coding for TRPM2. The intracellular solution used EDTA (10 mM) as a chelator for Ca$^{2+}$ and heavy metal ions. Moreover, we balanced the intracellular Ca$^{2+}$ concentration at 1 μM. H$_{2}$O$_{2}$ (10 mM) in the bath chamber was extracellularly added although ADPR (0.3 mM) and NAD$^{+}$ (1 mM) in pipette solution were intracellularly added. Using these conditions, the channel currents were evoked by the three stimulators. The time course of ADPR, NAD$^{+}$ and H$_{2}$O$_{2}$ effects was characterized by a delay of 0.6, 3.0 min and 2–5 min, respectively and a slow current induction reached a clear plateau with ADPR and NAD$^{+}$ although H$_{2}$O$_{2}$ currents continued to gain in amplitude over at least 15 min and it did not reach a clear plateau in many experiments. Furthermore, H$_{2}$O$_{2}$-induced a single-channel conductance in the current study; the first time that this has been resolved in CHO. The conductance of ADPR and H$_{2}$O$_{2}$ was 48.80 pS and 39.14 pS, respectively and the cells seem to be separately activated by ADPR and H$_{2}$O$_{2}$. In conclusion, we observed further support for a calcium influx pathway regulated separately by oxidative stress and ADPR in TRPM2 channels in transfected cells. A second novel result of the present study was that the TRPM2 channels were constitutionally activated by H$_{2}$O$_{2}$.
Starting Page	1256
Ending Page	1262
Page Count	7
File Format	PDF
ISSN	03643190
Journal	Neurochemical Research
Volume Number	33
Issue Number	7
e-ISSN	15736903
Language	English
Publisher	Springer US
Publisher Date	2008-02-08
Publisher Place	Boston
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	TRPM2 channels Single channel records Hydrogen peroxide ADP-ribose NAD$^{+}$ Neurology Biochemistry Neurosciences
Content Type	Text
Resource Type	Article
Subject	Medicine Biochemistry Cellular and Molecular Neuroscience

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