In Vitro Dose-Dependent Inhibition of the Intracellular Spontaneous Calcium Oscillations in Developing Hippocampal Neurons by Ketamine
In Vitro Dose-Dependent Inhibition of the Intracellular Spontaneous Calcium Oscillations in Developing Hippocampal Neurons by Ketamine
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DOI：
10.1371/journal.pone.0059804
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发表时间：
2013-03-28
期刊：
影响因子：
3.7
通讯作者：
Dong, Zhenming
中科院分区：
文献类型：
--
作者：
Huang, Lining;Liu, Yue;Dong, Zhenming
细胞内钙振荡的频率和幅度在空间和时间上的异常会影响神经细胞的正常生理功能。最近的研究表明，氯胺酮可以影响神经元的生长发育，甚至诱导神经元的凋亡死亡。本研究以分离的发育中的海马神经元为研究对象，通过负载Fluo-4的激光扫描共聚焦显微镜观察氯胺酮对发育中的海马神经元细胞内钙振荡的影响，进一步探讨其可能的机制。用半定量方法分析自发性钙振荡活动，在发育中的海马神经元观察到一种典型的钙振荡类型。此外，100 mU M的N-甲基-D-天冬氨酸(NMDA)使钙振荡幅度增加。40 mU M的MK801可抑制钙振荡的幅度和频率。结果表明，从30mM开始，随着氯胺酮浓度的增加，神经元钙振荡幅度逐渐降低。300mU氯胺酮对钙振荡频率的抑制作用具有统计学意义，3000mU氯胺酮可完全消除神经元钙振荡。1 mM氯胺酮处理的海马神经元分别给予100、300和1,000 mU N-甲基-D-天冬氨酸，以剂量依赖的方式恢复钙振荡的频率和幅度。事实上，1,000亩M-NMDA完全逆转了1 mM氯胺酮引起的钙振荡频率和幅度的下降。本研究揭示了氯胺酮通过NMDAR(NMDA受体)以剂量依赖的方式抑制发育中的海马神经元钙振荡的频率和幅度，这可能是氯胺酮对发育中的大鼠海马神经元毒性的一个潜在机制。
Spatial and temporal abnormalities in the frequency and amplitude of the cytosolic calcium oscillations can impact the normal physiological functions of neuronal cells. Recent studies have shown that ketamine can affect the growth and development and even induce the apoptotic death of neurons. This study used isolated developing hippocampal neurons as its study subjects to observe the effect of ketamine on the intracellular calcium oscillations in developing hippocampal neurons and to further explore its underlying mechanism using Fluo-4-loaded laser scanning confocal microscopy. Using a semi-quantitative method to analyze the spontaneous calcium oscillatory activities, a typical type of calcium oscillation was observed in developing hippocampal neurons. In addition, the administration of NMDA (N-Methyl-D-aspartate) at a concentration of 100 mu M increased the calcium oscillation amplitude. The administration of MK801 at a concentration of 40 mu M inhibited the amplitude and frequency of the calcium oscillations. Our results demonstrated that an increase in the ketamine concentration, starting from 30 mu M, gradually decreased the neuronal calcium oscillation amplitude. The inhibition of the calcium oscillation frequency by 300 mu M ketamine was statistically significant, and the neuronal calcium oscillations were completely eliminated with the administration of 3,000 mu M Ketamine. The administration of 100, 300, and 1,000 mu M NMDA to the 1 mM ketamine-pretreated hippocampal neurons restored the frequency and amplitude of the calcium oscillations in a dose-dependent manner. In fact, a concentration of 1,000 mu M NMDA completely reversed the decrease in the calcium oscillation frequency and amplitude that was induced by 1 mM ketamine. This study revealed that ketamine can inhibit the frequency and amplitude of the calcium oscillations in developing hippocampal neurons though the NMDAR (NMDA receptor) in a dose-dependent manner, which might highlight a possible underlying mechanism of ketamine toxicity on the rat hippocampal neurons during development.