Wu Chiping, Luk Wah Ping, Gillis Jesse, Skinner Frances, Zhang Liang
Toronto Western Research Institute, University Health Network, University of Toronto, Toronto, Ontario, Canada.
J Neurophysiol. 2005 Apr;93(4):2302-17. doi: 10.1152/jn.00806.2004. Epub 2004 Nov 10.
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.
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。