Inhibition of NMDA receptors induces delayed neuronal maturation and sustained proliferation of progenitor cells during neocortical development
Inhibition of NMDA receptors induces delayed neuronal maturation and sustained proliferation of progenitor cells during neocortical development
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DOI：
10.1002/jnr.10795
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发表时间：
2003-12-01
影响因子：
4.2
通讯作者：
Uchino, S
中科院分区：
文献类型：
--
作者：
Hirasawa, T;Wada, H;Uchino, S
为了阐明N-甲基-D-天冬氨酸（NMDA）受体在大脑新皮层发育早期的作用，我们研究了NMDA受体拮抗剂D（-）-2-氨基-5-膦酰基戊酸（D-APV）对胚胎第17天大鼠大脑新皮层切片培养和分离的原代培养细胞迁移和增殖的影响。用5-溴-2 '-脱氧尿苷（ErdU）的脉冲标记实验表明，在切片中慢性暴露于D-APV延迟神经元迁移。钙离子成像实验表明，功能性NMDA受体表达的神经元和处理D-APV延迟神经元成熟判断从亚基组成的NMDA受体亚型。使用BrdU脉冲标记的结果表明，暴露于D-APV 3天诱导新皮质切片中脑室区中增殖祖细胞数量明显增加。在原代培养物中暴露于D-APV也增加了祖细胞的增殖。D-APV对祖细胞增殖的影响可能是通过神经元细胞介导的。为了阐明D-APV诱导的祖细胞增殖增强的机制，我们通过逆转录-聚合酶链反应研究了Hes 1和Hes 5 mRNA在新皮质脑片脑室区的表达。暴露于D-APV 3天的组织中Hes 1和Hes 5 mRNA的表达高于未暴露的对照组织。这些结果表明，NMDA受体在神经元中表达的功能在神经元的迁移和成熟，祖细胞的增殖。(C)2003 Wiley-Liss，Inc.
To elucidate the role of N-methyl-D-aspartate (NMDA) receptors during the early stage of cerebral neocortical development, we investigated the effect of an NMDA receptor antagonist, D(-)-2-amino-5-phosphonopentanoic acid (D-APV), on cell migration and proliferation in slice cultures and dissociated primary cultures prepared from rat cerebral neocortex at embryonic Day 17. Pulse-labeling experiments with 5-bromo-2'-deoxyuridine (ErdU) showed that chronic exposure to D-APV in slices delayed neuronal migration. Calcium imaging experiments revealed that functional NMDA receptors were expressed in neurons and the treatment with D-APV delayed neuronal maturation judging from the subunit composition of NMDA receptor subtypes. The results using pulse-labeling with BrdU indicated that exposure to D-APV for 3 days induced a clear increase in the number of proliferating progenitor cells in the ventricular zone in neocortical slices. Exposure to D-APV in primary cultures also increased the proliferation of progenitor cells. The effect Of D-APV on progenitor cell proliferation was possibly mediated through neuronal cells. To elucidate the mechanism of enhanced progenitor cell proliferation induced by D-APV, we investigated expression of Hes1 and Hes5 mRNA in the ventricular zone of neocortical slices by reverse transcription-polymerase chain reaction. Tissue exposed to D-APV for 3 days showed higher expression of Hes1 and Hes5 mRNA than did unexposed control tissue. These results suggest that NMDA receptors expressed in neurons function in neuronal migration and maturation and in the proliferation of progenitor cells. (C) 2003 Wiley-Liss, Inc.