Methodological optimization of applying neuroactive agents for the study of locomotor-like activity in the mudpuppies (Necturus maculatus).
Methodological optimization of applying neuroactive agents for the study of locomotor-like activity in the mudpuppies (Necturus maculatus).
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DOI：
10.1016/j.jneumeth.2008.07.010
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发表时间：
2008-09-15
影响因子：
3
通讯作者：
Cheng, Jianguo
中科院分区：
文献类型：
--
作者：
Lavrov, Igor;Cheng, Jianguo
关键词：
我们比较了神经活性物质的给药方式和溶解神经活性物质的载体二甲基亚砜(DMSO)对体外运动样活性的影响。通过灌流，D-谷氨酸(0.3-0.9 mM)以10-25ml/min的灌流速度产生健壮的步行样活动。相反，团注相同或更高剂量的谷氨酸(0.1-1.5 mM)不能诱发任何节律性活动。灌流N-甲基-D-天冬氨酸受体拮抗剂AP-5可剂量依赖性地抑制D-谷氨酸诱导的行走样活动，并在20µM时完全阻断这种活动。相反，灌流AP-5在浓度达120µM时不能阻断行走样活动。同样，灌流AP-5可抑制行走样活动的启动，并在20µM时完全阻断行走样活动，而当灌流浓度达120µM时，AP-5不能阻断行走样活动。甘氨酸受体拮抗剂士的宁在3~5µM时阻断行走样活动。而它的推注将NMDA诱导的行走类活动改变为同步模式，而不是谷氨酸诱导的行走类活动。DMSO在1-10%(v/v)浓度范围内对类步行活动有明显的剂量依赖性影响。这些结果表明，神经活性物质的应用方式是决定运动神经控制实验结果的一个重要因素。此外，在数据解释中还应考虑DMSO对运动神经网络活动的剂量依赖效应。
We compared the effects of mode of delivery of neuroactive agents and the effects of Dimethyl sulfoxide (DMSO), a vehicle for dissolving neuroactive agents, on locomotor-like activity in vitro. By superfusion, D-glutamate (0.3 – 0.9 mM) produced robust walking-like activity at superfusion rates 10–25 ml/min. In contrast, bolus application of the same or higher doses of glutamate (0.1–1.5 mM) failed to induce any rhythmic activity. Superfusion with AP-5, a NMDA receptor antagonist, produced dose-dependent inhibition of the ongoing walking-like activity induced by D-glutamate and completely blocked the activity at 20 µM. In contrast, bolus application of AP-5 did not block the walking-like activity at concentrations up to 120 µM. Similarly, superfusion of AP-5 inhibited the initiation of walking-like activity and completely blocked the initiation at 20 µM, while bolus application of AP-5 failed to do so at concentrations up to 120 µM. Superfusion of strychnine, a glycine receptor antagonist, blocked the walking-like activity at concentrations of 3–5 µM, while its bolus application altered NMDA-induced, but not glutamate-induced, walking-like activity to a synchronized pattern. DMSO significantly affected the walking-like activity in a dose-dependent manner at concentrations ranging 1–10% (v/v). These results demonstrate that the way by which the neuroactive agents are applied is a significant factor that determines the outcome of experiments on the neural control of locomotion. Also, the dose-dependent effects of DMSO on the activity of neural networks for locomotion should be taken into account in data interpretation.
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