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JoVE Encyclopedia of Experiments
Neuroscience
海马网络动力学的光遗传学控制
海马网络动力学的光遗传学控制
Encyclopedia of Experiments
Neuroscience
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Encyclopedia of Experiments Neuroscience
Optogenetic Control of Hippocampal Network Dynamics

海马网络动力学的光遗传学控制

Protocol
345 Views
02:30 min
July 8, 2025
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Please note that some of the translations on this page are AI generated. Click here for the English version.

Transcript

从小鼠海马切片开始,该切片包含连接到转基因光敏中间神经元的靶锥体细胞。

将切片放入记录室中。

将

局部场电位或 LFP 电极放置在所需区域以记录 θ 振荡,这是一种有节奏的大脑活动。

使用微电极,修补附近的锥体细胞以记录其活动。

将光纤灯放置在以记录区域为中心的海马切片上方。

以

设定频率向切片传递蓝光脉冲,以激活中间神经元中的光敏离子通道。

激活导致正离子流入,导致去极化。

这会触发抑制性神经递质释放到锥体神经元上,从而降低兴奋性。

光刺激增强了 θ 振荡的同步和记录神经元的突触活动,表明神经节律的有效调节。

在此过程中,将LFP电极放置在CA1下区,并在PV中间神经元中表达蓝光敏感兴奋性视蛋白ChR2的小鼠的分离海马体中修补附近的锥体细胞。

将光纤光导放在海马准备上方,并将其置于记录区域的中心。使用来自 LED 光源的蓝光进行基因刺激,该刺激由以 θ 频率传递的 10 至 20 毫秒光脉冲或正弦波电压命令组成。在电流钳位中,表征自发 θ 振荡期间记录的细胞的活动。然后,启动刺激方案并记录光反应。

观察到记录的神经元中的场振荡和突触活动在光遗传学刺激期间变得越来越同步,并且PV细胞的节律性起搏导致对θ振荡的频率和功率的稳健控制。

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