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尺寸很重要:厚层小鼠海马切片中内在网络节律的产生

Size does matter: generation of intrinsic network rhythms in thick mouse hippocampal slices.

作者信息

Wu Chiping, Luk Wah Ping, Gillis Jesse, Skinner Frances, Zhang Liang

机构信息

Toronto Western Research Institute, University Health Network, University of Toronto, Toronto, Ontario, Canada.

出版信息

J Neurophysiol. 2005 Apr;93(4):2302-17. doi: 10.1152/jn.00806.2004. Epub 2004 Nov 10.

Abstract

Rodent hippocampal slices of < or = 0.5 mm thickness have been widely used as a convenient in vitro model since the 1970s. However, spontaneous population rhythmic activities do not consistently occur in this preparation due to limited network connectivity. To overcome this limitation, we develop a novel slice preparation of 1 mm thickness from adult mouse hippocampus by separating dentate gyrus from CA3/CA1 areas but preserving dentate-CA3-CA1 connectivity. While superfused in vitro at 32 or 37 degrees C, the thick slice exhibits robust spontaneous network rhythms of 1-4 Hz that originate from the CA3 area. Via assessing tissue O2, K+, pH, synaptic, and single-cell activities of superfused thick slices, we verify that these spontaneous rhythms are not a consequence of hypoxia and nonspecific experimental artifacts. We suggest that the thick slice contains a unitary circuitry sufficient to generate intrinsic hippocampal network rhythms and this preparation is suitable for exploring the fundamental properties and plasticity of a functionally defined hippocampal "lamella" in vitro.

摘要

自20世纪70年代以来,厚度小于或等于0.5毫米的啮齿动物海马切片作为一种便捷的体外模型被广泛使用。然而,由于网络连接有限,该制备物中并不总是会出现自发的群体节律活动。为了克服这一限制,我们通过将齿状回与CA3/CA1区域分离但保留齿状回-CA3-CA1连接性,开发了一种来自成年小鼠海马体的1毫米厚的新型切片制备方法。在32或37摄氏度的体外灌注条件下,厚切片表现出源自CA3区域的1-4赫兹的强烈自发网络节律。通过评估灌注厚切片的组织氧气、钾离子、酸碱度、突触和单细胞活动,我们证实这些自发节律并非缺氧和非特异性实验假象的结果。我们认为厚切片包含足以产生内在海马网络节律的单一电路,并且这种制备方法适合于在体外探索功能定义的海马“薄片”的基本特性和可塑性。

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