Ethanol preconditioning of rat cerebellar cultures targets NMDa receptors to the synapse and enhances peroxiredoxin 2 EXPRESSION Journal Article

Authors: Mitchell, R. M.; Tajuddin, N.; Campbell, E. M.; Neafsey, E. J.; Collins, M. A.
Article Title: Ethanol preconditioning of rat cerebellar cultures targets NMDa receptors to the synapse and enhances peroxiredoxin 2 EXPRESSION
Abstract: Epidemiological studies indicate that light-moderate alcohol (ethanol) consumers tend to have reduced risks of cognitive impairment and progression to dementia during aging. Exploring possible mechanisms, we previously found that moderate ethanol preconditioning (MEP, 20-30mM) of rat brain cultures for several days instigated neuroprotection against beta-amyloid peptides. Our biochemical evidence implicated the NMDA receptor (NMDAR) as a potential neuroprotective "sensor", specifically via synaptic NMDAR signaling. It remains unclear how ethanol modulates the receptor and its downstream targets to engender neuroprotection. Here we confirm with deconvolution microscopy that MEP of rat mixed cerebellar cultures robustly increases synaptic NMDAR localization. Phospho-activation of the non-receptor tyrosine kinases Src and Pyk2, known to be linked to synaptic NMDAR, is also demonstrated. Additionally, the preconditioning enhances levels of an antioxidant protein, peroxiredoxin 2 (Prx2), reported to be downstream of synaptic NMDAR signaling, and NMDAR antagonism with memantine (earlier found to abrogate MEP neuroprotection) blocks the Prx2 elevations. To further link Prx2 with antioxidant-based neuroprotection, we circumvented the ethanol preconditioning-NMDAR pathway by pharmacologically increasing Prx2 with the naturally-occurring cruciferous compound, 3H-1,2-dithiole-3-thione (D3T). Thus, D3T pretreatment elevated Prx2 expression to a similar extent as MEP, while concomitantly preventing beta-amyloid neurotoxicity; D3T also protected the cultures from hydrogen peroxide toxicity. The findings support a mechanism that couples synaptic NMDAR signaling, Prx2 expression and augmented antioxidant defenses in ethanol preconditioning-induced neuroprotection. That this mechanism can be emulated by a cruciferous vegetable constituent suggests that such naturally-occurring "neutraceuticals" may be useful in therapy for oxidative stress-related dementias.
Journal Title: Brain research
ISSN: 1872-6240; 0006-8993
Publisher: Elsevier Inc  
Date Published: 2016
Language: ENG
Notes: LR: 20160506; CI: Copyright (c) 2016; GR: R01 AA013568/AA/NIAAA NIH HHS/United States; GR: T32 AA013527/AA/NIAAA NIH HHS/United States; JID: 0045503; OTO: NOTNLM; 2015/07/23 [received]; 2016/02/29 [revised]; 2016/03/10 [accepted]; aheadofprint