Functional re-establishment of the perforant pathway in organotypic co-cultures on microelectrode arrays.
Co-cultures of entorhinal cortex (EC) and dentate gyrus (DG) explants are a useful model system to study the formation and stabilization of axonal projections. We adapted this model system to EC-DG co-cultures on microelectrode arrays (MEA) for the characterization of axonal projections on a functional level for days and weeks. EC and DG explants were placed on MEA to allow for the reconstitution of perforant pathway projections. Connections formed were characterized by morphological and electrophysiological analyses to verify characteristic features of perforant pathway signal transmission. Morphological analysis reveals proper projection of EC neurons into the molecular layer of the DG. Examination of synaptic transmission after high frequency stimulation imply unidirectional connections that used glutamate receptors of the AMPA/kainate type as main mediators of excitatory signal transmission. The system was evaluated by the introduction of the NCAM binding peptide C3d. In accordance with in vivo and in vitro experiments C3d modulated signal transmission by NCAM-related mechanisms resulting in morphological re-arrangements.
Hofmann
Frank
F
Naturwissenschaftliches und Medizinisches Institut an der Universität Tübingen, Markwiesenstr. 55, D-72770, Reutlingen, Germany.
Guenther
Elke
E
Hämmerle
Hugo
H
Leibrock
Cornelia
C
Berezin
Vladimir
V
Bock
Elisabeth
E
Volkmer
Hansjürgen
H
D018750:6-Cyano-7-nitroquinoxaline-2,3-dione / Q000494:pharmacology; D000200:Action Potentials / Q000502:physiology; D000818:Animals; D000831:Animals, Newborn; D001640:Bicuculline / Q000494:pharmacology; D001931:Brain Mapping; D002232:Carbocyanines / Q000378:metabolism; D018920:Coculture Techniques / Q000379:methods*; D015933:Complement C3d / Q000494:pharmacology; D018891:Dentate Gyrus / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004558:Electric Stimulation / Q000379:methods; D018728:Entorhinal Cortex / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004963:Estrenes / Q000494:pharmacology; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D018756:GABA Antagonists / Q000494:pharmacology; D007150:Immunohistochemistry / Q000379:methods; D007211:Indoles / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D051379:Mice; D008807:Mice, Inbred BALB C; D008839:Microelectrodes; D009416:Nerve Regeneration / Q000502:physiology; D009419:Nerve Tissue Proteins / Q000378:metabolism; D019006:Neural Cell Adhesion Molecules; D009924:Organ Culture Techniques; D019580:Perforant Pathway / Q000502:physiology*; D011743:Pyrimidines / Q000494:pharmacology; D011760:Pyrrolidinones / Q000494:pharmacology; D020127:Recovery of Function / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors; D014633:Valine / Q000031:analogs & derivatives* / Q000494:pharmacology
C035005:3,3'-dihexadecylindocarbocyanine; C412373:AG 1879; D002232:Carbocyanines; D004791:Enzyme Inhibitors; D004963:Estrenes; D018691:Excitatory Amino Acid Antagonists; D018756:GABA Antagonists; D007211:Indoles; D009419:Nerve Tissue Proteins; D019006:Neural Cell Adhesion Molecules; D011743:Pyrimidines; D011760:Pyrrolidinones; C033652:postsynaptic density proteins; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; C007293:DAPI; D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; C056639:2-amino-5-phosphopentanoic acid; D015933:Complement C3d; D014633:Valine; D001640:Bicuculline