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大鼠脑突触体中的天冬氨酸转运

Aspartate transport in synaptosomes from rat brain.

作者信息

Erecińska M, Wantorsky D, Wilson D F

出版信息

J Biol Chem. 1983 Aug 10;258(15):9069-77.

PMID:6874678
Abstract

Active transport of aspartate by neuronal synapses has been studied using preparations of synaptosomes from rat forebrain. The maximal accumulation of labeled D-aspartate [( Asp]i/[Asp]o) was observed to increase as the second power of the transmembrane electrical potential (measured by the potassium ion concentration gradient [( K+]i/[K+]o], which indicates that aspartate is transported with a net charge of +2. The aspartate gradient also increased as the second power of the sodium ion gradient, indicating that two Na+ are transported inward with each aspartate. Measurement of total intra- and extrasynaptosomal aspartate (L-aspartate) and the specific activities of each after adding labeled D-aspartate shows the presence of two intrasynaptosomal pools, a rapidly exchanging pool in which the L-aspartate is dependent on [Na+] and [K+] in the same manner as described for D-aspartate and a slowly exchanging pool essentially insensitive to [Na+] and [K+]. Maximal activation of uptake with 50 microM aspartate causes an alkalinization of the external medium which suggests that H+ is taken up with the aspartate (or OH- is ejected) to neutralize the negative charge of aspartate at physiological pH values. The data are consistent with the transport process being: Asp- + H+ + 2Na+ o = Asp- + H+ + 2Nai+. These stoichiometries can explain the energetics of both in vitro and in vivo uptake of aspartate by the high affinity, [Na+]- and [K+]-dependent system.

摘要

利用大鼠前脑突触体标本对神经元突触中天冬氨酸的主动转运进行了研究。观察到标记的D-天冬氨酸的最大积累量[(Asp)i/(Asp)o]随跨膜电位的平方增加(通过钾离子浓度梯度[(K+)i/(K+)o]测量),这表明天冬氨酸以+2的净电荷进行转运。天冬氨酸梯度也随钠离子梯度的平方增加,表明每转运一个天冬氨酸就有两个Na+向内转运。在加入标记的D-天冬氨酸后,对突触体内外总天冬氨酸(L-天冬氨酸)及其比活性的测量显示存在两个突触体内池,一个快速交换池,其中L-天冬氨酸对[Na+]和[K+]的依赖性与D-天冬氨酸描述的相同,另一个缓慢交换池基本上对[Na+]和[K+]不敏感。用50微摩尔天冬氨酸最大程度激活摄取会导致外部介质碱化,这表明H+与天冬氨酸一起被摄取(或OH-被排出)以中和生理pH值下天冬氨酸的负电荷。这些数据与转运过程一致:Asp- + H+ + 2Na+ o = Asp- + H+ + 2Nai+。这些化学计量关系可以解释高亲和力、依赖[Na+]和[K+]的系统在体外和体内摄取天冬氨酸的能量学。

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