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INNOVATIVE METHODOLOGY

Size Does Matter: Generation of Intrinsic Network Rhythms in Thick Mouse Hippocampal Slices

¹Toronto Western Research Institute, University Health Network, ²Department of Medicine, Division of Neurology, ³Department of Physiology, ⁴Institute of Biomaterials and Biomedical Engineering, and ⁵Epilepsy Research Program, University of Toronto, Toronto, Ontario, Canada

Submitted 6 August 2004; accepted in final form 7 November 2004

Rodent hippocampal slices of 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°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.

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