Long-term stimulation of mouse hippocampal slice culture on microelectrode array.
To understand mechanisms of information processing, development and degeneration of the central nervous system, simultaneous multisite recording and stimulation have become extremely helpful. We have further developed the innovative approach to record from intact neural networks using planar microelectrode arrays (MEAs) with 60 substrate-integrated nano-columnar electrodes. To allow for long-term stimulation, mouse hippocampal tissue slices were immobilized onto MEAs and permanently moved between the gas and medium phase in a specifically designed tilting incubator that made it possible to electrically contact up to 90 MEAs with 5400 electrodes. After 2-3 weeks in vitro, histochemical staining, the intracellular microinjection of the fluorescent dye Alexa and the recording of spontaneous activity revealed in vivo-like characteristics of the organotypically cultured tissue. The feasibility of long-term stimulation during culturing was demonstrated with a low frequency paradigm. 0.003 Hz stimulation over a 16 h period resulted in a significant decline of field potentials and population spikes in two identified hippocampal subregions. Control experiments revealed that this effect was not due to tissue detachment or to induced cell death. In summary, the novel technology promises to open a new avenue for analyzing regulatory interactions of neuronal activity, cell differentiation and gene expression during development and in diseases.
NMI Naturwissenschaftliches und Medizinisches Institut an der Universität Tübingen, Markwiesenstrasse 55, D-72770 Reutlingen, Germany.
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van Bergen
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Papanikolaou
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Schuker
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Möller
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Schlosshauer
D000200:Action Potentials / Q000502:physiology; D000818:Animals; D002470:Cell Survival / Q000502:physiology; D015194:Diffusion Chambers, Culture / Q000295:instrumentation / Q000379:methods*; D004558:Electric Stimulation / Q000295:instrumentation* / Q000379:methods*; D005260:Female; D005456:Fluorescent Dyes; D006624:Hippocampus / Q000166:cytology / Q000502:physiology*; D006834:Hydrazines; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008839:Microelectrodes / Q000592:standards*; D009924:Organ Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D013997:Time Factors
C417561:Alexa 488 hydrazide; D005456:Fluorescent Dyes; D006834:Hydrazines