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大鼠脑中NR2A和NR2B的发育表达、亚细胞定位及酪氨酸磷酸化

Developmental expression, subcellular localization, and tyrosine phosphorylation of NR2A and NR2B in the rat brain.

作者信息

Jin D H, Jung Y W, Ham S H, Ko B H, Moon I S

机构信息

Department of Anatomy, College of Medicine, Dongguk University, Kyongju, Korea.

出版信息

Mol Cells. 1997 Feb 28;7(1):64-71.

PMID:9085267
Abstract

We carried out quantitative analyses of the developmental expression, subcellular localization of the N-methyl-D-aspartate receptor subunit 2A (NR2A) and 2B (NR2B), and tyrosine phosphorylation of NR2B. Immunoblot analyses showed that NR2A was not detected during the embryonic period and the first postnatal week but its expression reached 63.90% of adult at P14 and continued to increase until the fourth week, reaching a maximum at P30 (110% of adult). The NR2B was detected from as early as E14 (2.65% of adult) and its expression was transiently elevated at birth (43.73% of adult), decreasing for the first postnatal week, and then increased again rapidly in the second week (105.45% of adult at P14) with a maximum at P30 (123.34% of adult). There were 2.26 +/- 0.40-fold more NR2B than NR2A proteins in the forebrain PSD fractions, and NR2A and NR2B were enriched 2.75 +/- 0.35 and 4.65 +/- 0.25 fold, respectively, in the synaptosome, and 13.75 +/- 0.80 and 16.04 +/- 0.25-fold, respectively, in the PSD fraction from brain homogenate. The tyrosine phosphorylation of NR2B reached an adult level at around birth declining in the first postnatal week but recovered to the adult level by the end of the second week, while the amount of the protein itself increased 2.28-fold after birth, indicating that only a fraction of the proteins are phosphorylated in vivo. Our results indicate that expression and tyrosine phosphorylation of NR2B might be important for NMDA receptor functions in embryonic and early postnatal development.

摘要

我们对N-甲基-D-天冬氨酸受体亚基2A(NR2A)和2B(NR2B)的发育表达、亚细胞定位以及NR2B的酪氨酸磷酸化进行了定量分析。免疫印迹分析表明,在胚胎期和出生后的第一周未检测到NR2A,但其表达在出生后第14天达到成年水平的63.90%,并持续增加直至第四周,在出生后第30天达到最大值(为成年水平的110%)。NR2B早在胚胎第14天就被检测到(为成年水平的2.65%),其表达在出生时短暂升高(为成年水平的43.73%),在出生后的第一周下降,然后在第二周再次迅速增加(出生后第14天为成年水平的105.45%),在出生后第30天达到最大值(为成年水平的123.34%)。在前脑突触后致密部(PSD)组分中,NR2B蛋白比NR2A蛋白多2.26±0.40倍,在突触体中,NR2A和NR2B分别富集2.75±0.35倍和4.65±0.25倍,在脑匀浆的PSD组分中分别富集13.75±0.80倍和16.04±0.25倍。NR2B的酪氨酸磷酸化在出生时左右达到成年水平,在出生后的第一周下降,但在第二周结束时恢复到成年水平,而蛋白质本身的量在出生后增加了2.28倍,这表明体内只有一部分蛋白质被磷酸化。我们的结果表明,NR2B的表达和酪氨酸磷酸化可能对胚胎期和出生后早期发育中的N-甲基-D-天冬氨酸受体功能很重要。

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