Biocompatibility of silicon-based arrays of electrodes coupled to organotypic hippocampal brain slice cultures.
In this study we examined the passive biocompatibility of a three-dimensional microelectrode array (MEA), designed to be coupled to organotypic brain slice cultures for multisite recording of electrophysiological signals. Hippocampal (and corticostriatal) brain slices from 1-week-old (and newborn) rats were grown for 4-8 weeks on the perforated silicon chips with silicon nitride surfaces and 40 microm sized holes and compared with corresponding tissue slices grown on conventional semiporous membranes. In terms of preservation of the basic cellular and connective organization, as visualized by Nissl staining, Timm sulphide silver-staining, microtubule-associated protein 2 (MAP2) and glial fibrillary acidic protein (GFAP) immunostaining, the slice cultures grown on chips did not differ from conventionally grown slice cultures. Neither were there any signs of astrogliosis or neurodegeneration around the upper recording part of the 47-microm-high platinum-tip electrodes. Slice cultures grown on a separate set of chips with platinum instead of silicon nitride surfaces also displayed normal MAP2 and GFAP immunostaining. The width of the GFAP-rich zone (glia limitans) at the bottom surface of the slice cultures was the same ( approximately 20 microm) in cultures grown on chips with silicon nitride and platinum surfaces and on conventional insert membranes. The slice cultures grown on chips maintained a normal, subfield differentiated susceptibility to the glutamate receptor agonist N-methyl-D-aspartate (NMDA) and the neurotoxin trimethyltin (TMT), as demonstrated by the cellular uptake of propidium iodide (PI), which was used as a reproducible and quantifiable marker for neuronal degeneration. We conclude that organotypic brain slice cultures can grow on silicon-based three-dimensional microelectrode arrays and develop normally with display of normal subfield differentiated susceptibilities to known excito- and neurotoxins. From this it is anticipated that the set-up, designed for recording of electrophysiological parameters, can be used for long-term studies of defined neuronal networks and provide valuable information on both normal, neurotoxicological and neuropathological conditions.
Anatomy and Neurobiology, Institute of Medical Biology, SDU-Odense University, Winsløwparken 21, 5000 C, Odense, Denmark. bkristensen@health.sdu.dk
B W
BW
Kristensen
J
J
Noraberg
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P
Thiébaud
M
M
Koudelka-Hep
J
J
Zimmer
D000818:Animals; D004396:Coloring Agents; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation / Q000379:methods*; D004581:Electronics / Q000295:instrumentation / Q000379:methods; D004594:Electrophysiology / Q000295:instrumentation / Q000379:methods; D018690:Excitatory Amino Acid Agonists / Q000633:toxicity; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D006624:Hippocampus / Q000737:chemistry / Q000187:drug effects / Q000502:physiology; D007150:Immunohistochemistry; D007202:Indicators and Reagents; D008422:Materials Testing; D008839:Microelectrodes; D008869:Microtubule-Associated Proteins / Q000032:analysis; D016202:N-Methylaspartate / Q000633:toxicity; D009924:Organ Culture Techniques / Q000379:methods*; D010984:Platinum; D011419:Propidium; D051381:Rats; D017208:Rats, Wistar; D017655:Silicon Compounds; D016622:Silver Staining; D014048:Tolonium Chloride; D014298:Trimethyltin Compounds / Q000633:toxicity
D004396:Coloring Agents; D018690:Excitatory Amino Acid Agonists; D005904:Glial Fibrillary Acidic Protein; D007202:Indicators and Reagents; D008869:Microtubule-Associated Proteins; D017655:Silicon Compounds; D014298:Trimethyltin Compounds; D014048:Tolonium Chloride; C046488:trimethyltin; D011419:Propidium; D010984:Platinum; D016202:N-Methylaspartate; C032734:silicon nitride