title: Ontogeny-recapitulating generation and tissue integration of ES cell-derived Purkinje cells.
abstract: Purkinje cells are the sole output neurons of the cerebellar cortex and their dysfunction causes severe ataxia. We found that Purkinje cells could be robustly generated from mouse embryonic stem (ES) cells by recapitulating the self-inductive signaling microenvironments of the isthmic organizer. The cell-surface marker Neph3 enabled us to carry out timed prospective selection of Purkinje cell progenitors, which generated morphologically characteristic neurons with highly arborized dendrites that expressed mature Purkinje cell-specific markers such as the glutamate receptor subunit GluRδ2. Similar to mature Purkinje cells, these neurons also showed characteristic spontaneous and repeated action potentials and their postsynaptic excitatory potentials were generated exclusively through nonNMDA glutamate receptors. Fetal transplantation of precursors isolated by fluorescence-activated cell sorting showed orthotopic integration of the grafted neurons into the Purkinje cell layer with their axons extending to the deep cerebellar nuclei and dendrites receiving climbing and parallel fibers. This selective preparation of bona fide Purkinje cells should aid future investigation of this important neuron.
authors.affiliation: Organogenesis and Neurogenesis Group, RIKEN Center for Developmental Biology, Kobe, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Muguruma
authors.forename: Ayaka
authors.initials: A
authors.lastname: Nishiyama
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Ono
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Miyawaki
authors.forename: Eri
authors.initials: E
authors.lastname: Mizuhara
authors.forename: Seiji
authors.initials: S
authors.lastname: Hori
authors.forename: Akira
authors.initials: A
authors.lastname: Kakizuka
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Obata
authors.forename: Yuchio
authors.initials: Y
authors.lastname: Yanagawa
authors.forename: Tomoo
authors.initials: T
authors.lastname: Hirano
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Sasai
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D002525:Cerebellar Cortex / Q000166:cytology / Q000196:embryology; D004622:Embryo, Mammalian; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D018507:Gene Expression Regulation, Developmental / Q000235:genetics / Q000502:physiology*; D005968:Glutamate Decarboxylase / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D007136:Immunoglobulins / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D008822:Mice, Transgenic; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D018408:Patch-Clamp Techniques / Q000379:methods; D011247:Pregnancy; D011689:Purkinje Cells / Q000502:physiology*; D013234:Stem Cells / Q000502:physiology*
chemical_list: D007136:Immunoglobulins; D008565:Membrane Proteins; C546601:Neph3 protein, mouse; D009419:Nerve Tissue Proteins; D049452:Green Fluorescent Proteins; D005968:Glutamate Decarboxylase; C402565:glutamate decarboxylase 1