西亚试剂：Ontogeny-recapitulating generation and tissue integration o

Ontogeny-recapitulating generation and tissue integration of ES cell–derived Purkinje cells
Keiko Muguruma,Ayaka Nishiyama,Yuichi Ono,Hiroyuki Miyawaki,Eri Mizuhara,Seiji Hori,Akira Kakizuka,Kunihiko Obata,Yuchio Yanagawa,Tomoo Hirano& Yoshiki Sasai

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.

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