title: Potential functional neural repair with grafted neural stem cells of early embryonic neuroepithelial origin.
abstract: The fate of grafted neuroepithelial stem cells in the normal mature brain environment was assessed both morphologically and electrophysiologically to confirm their feasibility in the functional repair of damaged neural circuitry. The neuroepithelial stem cells were harvested from the mesencephalic neural plate of transgenic green fluorescence protein-carrying rat embryos, and implanted into the normal adult rat striatum. The short- and long-term differentiation pattern of donor-derived cells was precisely monitored immunohistochemically. The functional abilities of the donor-derived cells and communication between them and the host were investigated using host-rat brain slices incorporating the graft with whole-cell patch-clamp recording. Vigorous differentiation of the neuroepithelial stem cells into mostly neurons was noted in the short-term with positive staining for tyrosine hydroxylase, suggesting that the donor-derived cells were exclusively following their genetically programmed fate, together with gamma-aminobutyric acid (GABA) and glutamate expression. In the long-term, the large number of donor-derived neurons was sustained, but the staining pattern showed expression of dopamine- and adenosine 3':5'-monophosphate-regulated phosphoprotein 32, suggesting that some neurons were following environmental cues, together with the appearance of some cholinergic neurons. Some donor-derived astrocytes were also seen in the graft. Many action potentials indicating the presence of both dopaminergic and non-dopaminergic patterns could be elicited and recorded in the donor-derived neurons in addition to spontaneous glutamatergic and GABAergic post-synaptic currents which were strongly shown to be of host origin. Neuroepithelial stem cells are therefore an attractive candidate as a source of donor material for intracerebral grafting in functional repair.
authors.affiliation: Department of Neurosurgery, School of Medicine, Keio University, Tokyo 160-8582, Japan. koichi@sc.itc.keio.ac.jp
authors.forename: Koichi
authors.initials: K
authors.lastname: Uchida
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Momiyama
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Okano
authors.forename: Michisuke
authors.initials: M
authors.lastname: Yuzaki
authors.forename: Amane
authors.initials: A
authors.lastname: Koizumi
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Mine
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kawase
mesh_terms: D018750:6-Cyano-7-nitroquinoxaline-2,3-dione / Q000494:pharmacology; D000818:Animals; D030801:Animals, Genetically Modified; D001640:Bicuculline / Q000494:pharmacology; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D003342:Corpus Striatum / Q000166:cytology / Q000502:physiology / Q000601:surgery; D051937:Dopamine and cAMP-Regulated Phosphoprotein 32; D004347:Drug Interactions; D051959:ELAV Proteins; D004558:Electric Stimulation / Q000379:methods; D004622:Embryo, Mammalian; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D018756:GABA Antagonists / Q000494:pharmacology; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D005968:Glutamate Decarboxylase / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000096:biosynthesis; D007150:Immunohistochemistry / Q000379:methods; D066298:In Vitro Techniques; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D026901:Membrane Transport Proteins / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D046569:Neuroepithelial Cells / Q000502:physiology*; D009474:Neurons / Q000187:drug effects / Q000502:physiology* / Q000528:radiation effects; D018408:Patch-Clamp Techniques / Q000379:methods; D010750:Phosphoproteins / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D020127:Recovery of Function / Q000502:physiology*; D033581:Stem Cell Transplantation / Q000379:methods*; D013234:Stem Cells / Q000502:physiology*; D013997:Time Factors; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism; D050599:Vesicular Glutamate Transport Protein 2
chemical_list: D051937:Dopamine and cAMP-Regulated Phosphoprotein 32; D051959:ELAV Proteins; D018691:Excitatory Amino Acid Antagonists; D018756:GABA Antagonists; D005904:Glial Fibrillary Acidic Protein; C489787:MAP2 protein, rat; D026901:Membrane Transport Proteins; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; C493475:Slc17a6 protein, rat; D050599:Vesicular Glutamate Transport Protein 2; D049452:Green Fluorescent Proteins; D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; D014446:Tyrosine 3-Monooxygenase; D002795:Choline O-Acetyltransferase; D005968:Glutamate Decarboxylase; D001640:Bicuculline