Zhang Xuying, Nadler J Victor
Department of Pharmacology, Box 3813, Duke University Medical Center, Durham, NC 27710, USA.
Brain Res. 2009 Oct 27;1295:13-20. doi: 10.1016/j.brainres.2009.07.104. Epub 2009 Aug 5.
Aspartate satisfies all the criteria normally required for identification of a CNS neurotransmitter. Nevertheless, little electrophysiological evidence supports the existence of aspartate transmission. In studies with rat hippocampal synaptosomes, chemically evoked aspartate release differed from glutamate release in its relative sensitivity to increased Ca(2+) concentration outside the presynaptic active zones, inefficient coupling to P/Q-type Ca(2+) channels, sensitivity to KB-R7943, and resistance to native Clostridial toxins. We took advantage of these differences to search for a potential aspartate-mediated response at Schaffer collateral synapses in organotypic hippocampal slice cultures. The slice cultures were pretreated with botulinum neurotoxin C (BoNT/C) to eliminate most of the glutamate release so that an expectedly smaller aspartate-like component of the compound EPSC could be detected by whole cell patch clamp recording. In control cultures, NMDA receptor activation accounted for only 18% of the evoked EPSC and an NR2B-selective antagonist reduced the NMDA receptor-mediated component by only 20%. Block of P/Q-type Ca(2+) channels essentially eliminated the response and 0.1 muM KB-R7943 had no significant effect. In BoNT/C-pretreated cultures, however, NMDA receptor activation accounted for 77% of the evoked EPSC and an NR2B-selective antagonist reduced the NMDA receptor-mediated component by 57%. Block of P/Q-type Ca(2+) channels reduced the response by only 28%, but 0.1 muM KB-R7943 reduced it by 45%. These results suggest that part of the Schaffer collateral synaptic response has pharmacological properties similar to those of synaptosomal aspartate release and may therefore be mediated at least partly by released aspartate.
天冬氨酸满足通常用于鉴定中枢神经系统神经递质的所有标准。然而,几乎没有电生理证据支持天冬氨酸传递的存在。在对大鼠海马突触体的研究中,化学诱发的天冬氨酸释放与谷氨酸释放在对突触前活性区外增加的Ca(2+)浓度的相对敏感性、与P/Q型Ca(2+)通道的低效偶联、对KB-R7943的敏感性以及对天然梭菌毒素的抗性方面存在差异。我们利用这些差异在器官型海马切片培养物的Schaffer侧支突触中寻找潜在的天冬氨酸介导的反应。用肉毒杆菌神经毒素C(BoNT/C)预处理切片培养物以消除大部分谷氨酸释放,从而可以通过全细胞膜片钳记录检测到复合兴奋性突触后电流(EPSC)中预期较小的类似天冬氨酸的成分。在对照培养物中,NMDA受体激活仅占诱发的EPSC的18%,并且NR2B选择性拮抗剂仅将NMDA受体介导的成分减少了20%。阻断P/Q型Ca(2+)通道基本上消除了反应,而0.1μM KB-R7943没有显著影响。然而,在BoNT/C预处理的培养物中,NMDA受体激活占诱发的EPSC的77%,并且NR2B选择性拮抗剂将NMDA受体介导的成分减少了57%。阻断P/Q型Ca(2+)通道仅使反应减少了28%,但0.1μM KB-R7943使其减少了45%。这些结果表明,Schaffer侧支突触反应的一部分具有与突触体天冬氨酸释放相似的药理学特性,因此可能至少部分由释放的天冬氨酸介导。