Keller B U, Hollmann M, Heinemann S, Konnerth A
Max Planck Institut für biophysikalische Chemie, Göttingen, Germany.
EMBO J. 1992 Mar;11(3):891-6. doi: 10.1002/j.1460-2075.1992.tb05127.x.
Excitatory synaptic transmission in the central nervous system (CNS) is mediated by three major classes of glutamate receptors, namely the ionotropic NMDA (N-Methyl-D-Aspartate) and KA/AMPA (kainate/alpha-amino-3-hydroxyl-5-methylisoxazole-4-propionic acid) receptors and the metabotropic receptor type. Among the ionotropic receptors, NMDA receptors are thought to mediate their physiological response mainly through the influx of extracellular calcium, while KA/AMPA receptor channels are mainly thought to carry the influx of monovalent cations. Recently, we have challenged this view by showing that cloned KA/AMPA receptor subunits GluR1 and GluR3 form ion channels which are permeable to calcium. We now directly demonstrate large increases in intracellular calcium concentrations induced by calcium fluxes through KA/AMPA receptor channels in solutions with physiological calcium concentrations. Calcium fluxes were observed through glutamate receptor channels composed of the subunits GluR1 and GluR3, which are both abundantly present in various types of central neurones. The calcium influx was fluorometrically monitored in Xenopus oocytes injected with the calcium indicator dye fura-2. Bath application of the membrane permeable analogue of adenosine cyclic monophosphate (cAMP) potentiated the current and also the flux of calcium through open KA/AMPA receptor channels. Further pharmacological experiments suggested that this effect was mediated by the activation of protein kinase A. Our results provide a molecular interpretation for the function of calcium permeable KA/AMPA receptor channels in neurones and identify two of the subunits of the KA/AMPA receptor channel which are regulated by the cAMP dependent second messenger system.
中枢神经系统(CNS)中的兴奋性突触传递由三类主要的谷氨酸受体介导,即离子型N-甲基-D-天冬氨酸(NMDA)受体和海人藻酸/α-氨基-3-羟基-5-甲基异恶唑-4-丙酸(KA/AMPA)受体以及代谢型受体。在离子型受体中,NMDA受体被认为主要通过细胞外钙的内流介导其生理反应,而KA/AMPA受体通道主要被认为携带单价阳离子的内流。最近,我们对这一观点提出了挑战,表明克隆的KA/AMPA受体亚基GluR1和GluR3形成了对钙通透的离子通道。我们现在直接证明,在具有生理钙浓度的溶液中,通过KA/AMPA受体通道的钙通量会导致细胞内钙浓度大幅增加。通过由亚基GluR1和GluR3组成的谷氨酸受体通道观察到了钙通量,这两种亚基在各种类型的中枢神经元中都大量存在。在注射了钙指示剂染料fura-2的非洲爪蟾卵母细胞中,通过荧光法监测钙内流。膜通透性环磷酸腺苷(cAMP)类似物的浴槽给药增强了电流以及通过开放的KA/AMPA受体通道的钙通量。进一步的药理学实验表明,这种效应是由蛋白激酶A的激活介导的。我们的结果为神经元中钙通透KA/AMPA受体通道的功能提供了分子解释,并鉴定了受cAMP依赖性第二信使系统调节的KA/AMPA受体通道的两个亚基。