title: Transplantation of motoneurons derived from MASH1-transfected mouse ES cells reconstitutes neural networks and improves motor function in hemiplegic mice.
abstract: Mouse embryonic stem (ES) cells were transfected with a MASH1 expression vector and G418-resistant cells were selected. The MASH1-transfected cells became neuron-like appearance and expressed betaIIItubulin and panNCAM. Glial fibrillary acidic protein (GFAP) and galactocerebroside (GalC)-expressing cells were rarely detected. Half of the neural cells differentiated into the Islet1+ motoneuron lineage. Thus, we obtained motoneuron lineage-enriched neuronal cells by transfection of ES cells with MASH1. A hemiplegic model of mice was developed by cryogenic injury of the motor cortex, and motoneuron lineage-enriched neuronal cells were transplanted underneath the injured motor cortex neighboring the periventricular region. The motor function of the recipients was assessed by a beam walking and rotarod tests, whereby the results gradually improved, but little improvement was observed in vehicle injected control mice. We found that the grafted cells not only remained close to the implantation site, but also exhibited substantial migration, penetrating into the damaged lesion in a directed manner up to the cortical region. Grafted neuronal cells that had migrated into the cortex were elongated axon-positive for neurofilament middle chain (NFM). Synaptophysin immunostaining showed a positive staining pattern around the graft, suggesting that the transplanted neurons interacted with the recipient neurons to form a neural network. Our study suggests that the motoneuron lineage can be induced from ES cells, and grafted cells adapt to the host environment and can reconstitute a neural network to improve motor function of a paralyzed limb.
authors.affiliation: Department of Immunology and Medicine, St. Marianna University School of Medicine, 2-16-1, Sugao, Miyamae, Kawasaki, Kanagawa 216-8511, Japan.
authors.forename: Ritsuko
authors.initials: R
authors.lastname: Ikeda
authors.forename: Manae S
authors.initials: MS
authors.lastname: Kurokawa
authors.forename: Shunmei
authors.initials: S
authors.lastname: Chiba
authors.forename: Hideshi
authors.initials: H
authors.lastname: Yoshikawa
authors.forename: Takuo
authors.initials: T
authors.lastname: Hashimoto
authors.forename: Mamoru
authors.initials: M
authors.lastname: Tadokoro
authors.forename: Noboru
authors.initials: N
authors.lastname: Suzuki
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins; D001930:Brain Injuries / Q000378:metabolism / Q000503:physiopathology / Q000628:therapy*; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage / Q000235:genetics; D002465:Cell Movement / Q000235:genetics; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*; D004195:Disease Models, Animal; D005260:Female; D006085:Graft Survival / Q000502:physiology; D006429:Hemiplegia / Q000378:metabolism / Q000503:physiopathology / Q000628:therapy*; D051379:Mice; D009044:Motor Cortex / Q000293:injuries / Q000503:physiopathology; D009046:Motor Neurons / Q000378:metabolism / Q000637:transplantation*; D009069:Movement Disorders / Q000378:metabolism / Q000503:physiopathology / Q000628:therapy; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D009415:Nerve Net / Q000166:cytology / Q000502:physiology*; D019006:Neural Cell Adhesion Molecules / Q000378:metabolism; D016900:Neurofilament Proteins / Q000378:metabolism; D020127:Recovery of Function / Q000502:physiology; D016708:Synaptophysin / Q000378:metabolism; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection / Q000379:methods; D016896:Treatment Outcome; D014404:Tubulin / Q000378:metabolism
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D004268:DNA-Binding Proteins; D019006:Neural Cell Adhesion Molecules; D016900:Neurofilament Proteins; D016708:Synaptophysin; D014157:Transcription Factors; D014404:Tubulin; C499317:Ascl1 protein, mouse; C061195:neurofilament protein M