Foster A C, Mena E E, Fagg G E, Cotman C W
J Neurosci. 1981 Jun;1(6):620-5. doi: 10.1523/JNEUROSCI.01-06-00620.1981.
The binding of the putative excitatory transmitters glutamate (Glu) and aspartate (Asp) was measured in various subcellular fractions in order to assess their degree of localization in synaptic junctions (SJs). For both ligands, specific binding levels increased in the order, whole particulate membranes--crude mitochondrial pellet membranes (P2)--synaptic plasma membranes (SPM), and were highly enriched in SJs, with values approximately 9 times greater than the values in whole particulate membranes. The recovery of binding sites in SJs suggested that the majority of sites in SPMs were junctional in nature. Specific binding sites were found also in other subcellular fractions, such as microsomal membranes, "light" SPMs, and mitochondrial membranes. Sodium ions were able to stimulate the specific binding of both ligands (Asp greater than Glu), the magnitude of the effect between subcellular fractions being in the order, whole particulate membranes greater than P2 greater than SPM. This effect was absent in SJs. Calcium and magnesium ions also enhanced the binding (Glu greater than Asp) in the order, whole particulate membranes greater than or equal to P2 greater than or equal to SPM greater than SJ. The results indicate that Glu and Asp binding sites have a specific synaptic localization and support a role for Glu and Asp receptors in synaptic transmission.
为了评估假定的兴奋性递质谷氨酸(Glu)和天冬氨酸(Asp)在突触连接(SJ)中的定位程度,我们测定了它们在各种亚细胞组分中的结合情况。对于这两种配体,特异性结合水平按以下顺序增加:全颗粒膜——粗线粒体沉淀膜(P2)——突触质膜(SPM),并且在SJ中高度富集,其值比全颗粒膜中的值大约高9倍。SJ中结合位点的回收率表明,SPM中的大多数位点本质上是连接性的。在其他亚细胞组分中也发现了特异性结合位点,如微粒体膜、“轻”SPM和线粒体膜。钠离子能够刺激这两种配体的特异性结合(Asp大于Glu),亚细胞组分之间的效应大小顺序为:全颗粒膜大于P2大于SPM。这种效应在SJ中不存在。钙和镁离子也按以下顺序增强结合(Glu大于Asp):全颗粒膜大于或等于P2大于或等于SPM大于SJ。结果表明,Glu和Asp结合位点具有特定的突触定位,并支持Glu和Asp受体在突触传递中的作用。