Cross-talk between α7 nAchR and NMDAR revealed by protein profiling.
Functional regulation of NMDA receptor (NMDAR) by the activation of α7 nicotinic acetylcholine receptor (α7nAChR) has been reported, although the molecular signaling pathway underlying this process remains largely unknown. We employed a label-free quantitative proteomics approach to identify potential intracellular molecules and pathways that might be involved in the functional cross-talk between NMDAR and α7nAChR. 43 proteins showed significantly expression changes after choline treatment in which 35 out of 43 proteins was significantly altered by co-treatment with NMDA. Western blot analysis verified proteins expression determined by LC-MS. Furthermore, protein expression in vivo in neurons from fetal rats were visualized and quantified by Confocal microscopy,which showed consistency of relative changes of AHA-1 expressionmeasured by LC-MS and Western blot. Biological network analysis of differently expressed proteins found 21 kind of biological pathways involved. Of those pathways, 6 pathways were directly involved in regulation of neurotransmitters. Lastly, the levels of neurotransmitters (dopamine, glutamate, GABA and 5-HT) were measured by HPLC-ECD. Co-treatment choline/NMDA significantly enhances the release of dopamine, glutamate and GABA and dramatically decrease 5-HT to only 65% of control level, which is consist with this protein interaction network analysis, providing an additional evidence for the cross-talk between NMDAR and α7nAChR.
Campbell Research Institute, Centre for Addiction and Mental Health, Toronto, Ontario M5T 1R8, Canada.
Hailong
H
Zhang
Campbell Research Institute, Centre for Addiction and Mental Health, Toronto, Ontario M5T 1R8, Canada; Medical and Forensic College, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi, 710061, PR China.
Tao
T
Li
Campbell Research Institute, Centre for Addiction and Mental Health, Toronto, Ontario M5T 1R8, Canada; Department of Psychiatry, University of Toronto, Toronto, Ontario M5T 1R8, Canada.
Shupeng
S
Li
Campbell Research Institute, Centre for Addiction and Mental Health, Toronto, Ontario M5T 1R8, Canada; Department of Psychiatry, University of Toronto, Toronto, Ontario M5T 1R8, Canada. Electronic address: f.liu.a@utoronto.ca.
Fang
F
Liu
D000818:Animals; D003198:Computer Simulation; D020869:Gene Expression Profiling / Q000379:methods*; D006624:Hippocampus / Q000378:metabolism*; D008297:Male; D008954:Models, Biological; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D025941:Protein Interaction Mapping / Q000379:methods; D051381:Rats; D020318:Rats, Long-Evans; D020239:Receptor Cross-Talk; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*; D064569:alpha7 Nicotinic Acetylcholine Receptor / Q000378:metabolism*
D009419:Nerve Tissue Proteins; D016194:Receptors, N-Methyl-D-Aspartate; D064569:alpha7 Nicotinic Acetylcholine Receptor
Biological pathway; Label-free quantitative proteomics approach; NMDAR; Neurotransmitters; Protein–protein interaction; α7nAchR