Hyper-SUMOylation of K+ Channels in Sudden Unexplained Death in Epilepsy: Isolation and Primary Culture of Dissociated Hippocampal Neurons from Newborn Mice for Subcellular Localization
Hyper-SUMOylation of K+ Channels in Sudden Unexplained Death in Epilepsy: Isolation and Primary Culture of Dissociated Hippocampal Neurons from Newborn Mice for Subcellular Localization
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癫痫不明原因猝死中 K( ) 通道的过度 SUMOylation：新生小鼠分离海马神经元的分离和原代培养，用于亚细胞定位。
DOI：
10.1007/978-1-4939-7362-0_6
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发表时间：
2018-01-01
期刊：
影响因子：
--
通讯作者：
Qi, Yitao
中科院分区：
文献类型：
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作者：
Chen, Xu;Zhang, Shanshan;Qi, Yitao
大鼠和小鼠海马神经元的生理特性被广泛研究，特别是因为海马参与学习、记忆和神经功能。海马神经元的原代培养通常用于发现神经生物学中的细胞和分子机制。通过分离和培养单个海马神经元，神经科学家能够在单个细胞和单个突触水平上研究神经元的活动，并使用各种生物化学技术分析与细胞结构，细胞运输和单个蛋白质亚细胞定位或蛋白质-蛋白质相互作用相关的特性。从这些研究中得出的结论对于测试与记忆，学习和神经功能相关的理论至关重要。在这里，我们将描述如何分离和培养原代海马细胞从新生小鼠。海马可以在短至2分钟内从新生小鼠中分离，并且细胞培养物可以维持长达2周，然后准备用于研究K+通道蛋白的亚细胞定位以及与SUMO特异性蛋白酶2（SENP 2）的相互作用。该方法为从小鼠海马组织中培养神经细胞提供了一种快速、有效的技术，为神经学研究中的亚细胞定位或蛋白质间相互作用的免疫细胞化学检测提供了保证。
The physiological characteristics of rat and murine hippocampal neurons are widely studied, especially because of the involvement of the hippocampus in learning, memory, and neurological functions. Primary cultures of hippocampal neurons are commonly used to discover cellular and molecular mechanisms in neurobiology. By isolating and culturing individual hippocampal neurons, neuroscientists are able to investigate the activity of neurons at the individual cell and single synapse level, and to analyze properties related to cellular structure, cellular trafficking, and individual protein subcellular localization or protein-protein interaction using a variety of biochemical techniques. Conclusions addressed from such research are critical for testing theories related to memory, learning, and neurological functions. Here, we will describe how to isolate and culture primary hippocampal cells from newborn mice. The hippocampus may be isolated from newborn mice in as short as 2 min, and the cell cultures can be maintained for up to 2 weeks, and then ready for investigation of subcellular localization of K+ channel proteins and interaction with SUMO-specific protease 2 (SENP2). The protocol provides a fast and efficient technique for the culture of neuronal cells from mouce hippocampal tissue, and will ensure the immunocytochemistry detection of subcellular localization or protein-protein interactions in neurological research.