Dual Effect of Methylglyoxal on the Intracellular Ca2+ Signaling and Neurite Outgrowth in Mouse Sensory Neurons
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The formation of advanced glycation end products is one of the major factors involved in diabetic neuropathy, aging, and neurodegenerative diseases. Reactive carbonyl compounds, such as methylglyoxal (MG), play a key role in cross-linking to various proteins in the extracellular matrix, especially in neurons, which have a high rate of oxidative metabolism. The MG effect was tested on dorsal root ganglia primary neurons in cultures from adult male Balb/c mice. Lower MG doses contribute to an increased adherence of neurons on their support and an increased glia proliferation, as proved by MTS assay and bright-field microscopy. Time-lapse fluorescence microscopy by Fura-2 was performed for monitoring the relative fluorescence ratio changes (ΔR/R 0) upon depolarization and immunofluorescence staining for quantifying the degree of neurites extension. The relative change in fluorescence ratio modifies the amplitude and dispersion depending on the subtype of sensory neurons, the medium-sized neurons are more sensitive to MG treatment when compared to small ones. Low MG concentrations (0–150 μM) increase neuronal viability, excitability, and the capacity of neurite extension, while higher concentrations (250–750 μM) are cytotoxic in a dose-dependent manner. In our opinion, MG could be metabolized by the glyoxalase system inside sensory neurons up to a threshold concentration, afterwards disturbing the cell equilibrium. Our study points out that MG has a dual effect concentration dependent on the neuronal viability, excitability, and neurite outgrowth, but only the excitability changes are soma-sized dependent. In conclusion, our data may partially explain the distinct neuronal modifications in various neurodegenerative pathologies.
KeywordsMethylglyoxal Viability Time-lapse fluorescence microscopy Neurite outgrowth Peripheral sensory neurons
This work was supported by the national grant PNII 41-074/2007 from the Romanian Ministry of Research. A great thanks to the technicians Cornelia Dragomir, Geanina Haralambie, and Constantin Radulescu for a constant help during the experiments.
- Engel MA, Leffler A, Niedermirtl F, Babes A, Zimmermann K, Filipović MR, Izydorczyk I, Eberhardt M, Kichko TI, Mueller-Tribbensee SM, Khalil M, Siklosi N, Nau C, Ivanović-Burmazović I, Neuhuber WL, Becker C, Neurath MF, Reeh PW (2011) TRPA1 and substance P mediate colitis in mice. Gastroenterology 141(4):1346–1358PubMedCrossRefGoogle Scholar
- Kikuchi S, Shinpo K, Moriwaka F, Makita Z, Miyata T, Tashiro K (1999) Neurotoxicity of methylglyoxal and 3-deoxyglucosone on cultured cortical neurons: synergism between glycation and oxidative stress, possibly involved in neurodegenerative diseases. J Neurosci Res 57(2):280–289PubMedCrossRefGoogle Scholar
- Koivisto A, Hukkanen M, Saarnilehto M, Chapman H, Kuokkanen K, Wei H, Viisanen H, Akerman KE, Lindstedt K, Pertovaara A (2012) Inhibiting TRPA1 ion channel reduces loss of cutaneous nerve fiber function in diabetic animals: Sustained activation of the TRPA1 channel contributes to the pathogenesis of peripheral diabetic neuropathy. Pharmacol Res 65(1):149–158PubMedCrossRefGoogle Scholar
- Lupachyk S, Shevalye H, Maksimchyk Y, Drel VR, Obrosova IG (2011) PARP inhibition alleviates diabetes-induced systemic oxidative stress and neural tissue 4-hydroxynonenal adduct accumulation: correlation with peripheral nerve function. Free Radic Biol Med 50(10):1400–1409PubMedCrossRefGoogle Scholar
- WO/2010/136182 WIPO patent application, Newroth P, Bierhaus A, Fleming T (2010) Methylglyoxal-scavenging compounds and their use for the prevention and treatment of pain and/or hyperalgesiaGoogle Scholar