Zhao Xia, Han Chao, Zeng Zhiwen, Liu Linlin, Wang Haitao, Xu Jiangping, Feng Zhong-Ping, Little Peter J, Quirion Remi, Zheng Wenhua
Faculty of Health Sciences, University of Macau, Taipa, Macau, China.
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center and School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou, China.
Oxid Med Cell Longev. 2020 Aug 11;2020:5173184. doi: 10.1155/2020/5173184. eCollection 2020.
Glutamate-induced neurotoxicity is involved in various neuronal diseases, such as Alzheimer's disease. We have previously reported that glutamate attenuated the survival signaling of insulin-like growth factor-1 (IGF-1) by N-methyl-D-aspartate receptors (NMDARs) in cultured cortical neurons, which is viewed as a novel mechanism of glutamate-induced neurotoxicity. However, the phosphorylation sites of IGF-1 receptor (IGF-1R) affected by glutamate remain to be elucidated, and importantly, which subtype of NMDARs plays a major role in attenuating the prosurvival effect of IGF-1 is still unknown. In the present study, glutamate was found to attenuate the tyrosine phosphorylation of the IGF-1R and the prosurvival effect of IGF-1 in primary cultured cortical neurons. NMDAR inhibitors, MK801 and AP-5, blocked the inhibitory effect of glutamate on the phosphorylation of IGF-1R and increased cell survival, while DNQX, LY341495, and CPCCOEt had no effect. Interestingly, we found that glutamate decreased the phosphorylation of tyrosine residues 1131, 1135/1136, 1250/1251, and 1316, while it had no effect on tyrosine 950 in cortical neurons. Moreover, using specific antagonists and siRNA to downregulate individual NMDAR subunits, we found that the activation of NR2B-containing NMDARs was essential for glutamate to inhibit IGF-1 signaling. These findings indicate that the glutamate-induced attenuation of IGF-1 signaling is mediated by NR2B-containing NMDARs. Our study also proposes a novel mechanism of altering neurotrophic factor signaling by the activation of NMDARs.
谷氨酸诱导的神经毒性与多种神经元疾病有关,如阿尔茨海默病。我们之前报道过,在培养的皮质神经元中,谷氨酸通过N-甲基-D-天冬氨酸受体(NMDARs)减弱胰岛素样生长因子-1(IGF-1)的生存信号,这被视为谷氨酸诱导神经毒性的一种新机制。然而,受谷氨酸影响的IGF-1受体(IGF-1R)的磷酸化位点仍有待阐明,重要的是,哪种NMDAR亚型在减弱IGF-1的促生存作用中起主要作用仍不清楚。在本研究中,发现谷氨酸可减弱原代培养皮质神经元中IGF-1R的酪氨酸磷酸化以及IGF-1的促生存作用。NMDAR抑制剂MK801和AP-5可阻断谷氨酸对IGF-1R磷酸化的抑制作用并提高细胞存活率,而DNQX、LY341495和CPCCOEt则无此作用。有趣的是,我们发现谷氨酸可降低皮质神经元中酪氨酸残基1131、1135/1136、1250/1251和1316的磷酸化水平,但对酪氨酸950无影响。此外,使用特异性拮抗剂和小干扰RNA(siRNA)下调单个NMDAR亚基,我们发现含NR2B的NMDARs的激活对于谷氨酸抑制IGF-1信号至关重要。这些发现表明,谷氨酸诱导的IGF-1信号减弱是由含NR2B的NMDARs介导的。我们的研究还提出了一种通过激活NMDARs改变神经营养因子信号的新机制。