title: Visualization, direct isolation, and transplantation of midbrain dopaminergic neurons.
abstract: To visualize and isolate live dopamine (DA)-producing neurons in the embryonic ventral mesencephalon, we generated transgenic mice expressing green fluorescent protein (GFP) under the control of the rat tyrosine hydroxylase gene promoter. In the transgenic mice, GFP expression was observed in the developing DA neurons containing tyrosine hydroxylase. The outgrowth and cue-dependent guidance of GFP-labeled axons was monitored in vitro with brain culture systems. To isolate DA neurons expressing GFP from brain tissue, cells with GFP fluorescence were sorted by fluorescence-activated cell sorting. More than 60% of the sorted GFP(+) cells were positive for tyrosine hydroxylase, confirming that the population had been successfully enriched with DA neurons. The sorted GFP(+) cells were transplanted into a rat model of Parkinson's disease. Some of these cells survived and innervated the host striatum, resulting in a recovery from Parkinsonian behavioral defects. This strategy for isolating an enriched population of DA neurons should be useful for cellular and molecular studies of these neurons and for clinical applications in the treatment of Parkinson's disease.
authors.affiliation: Division of Neuroanatomy (D12), Department of Neuroscience, Biomedical Research Center, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Sawamoto
authors.forename: N
authors.initials: N
authors.lastname: Nakao
authors.forename: K
authors.initials: K
authors.lastname: Kobayashi
authors.forename: N
authors.initials: N
authors.lastname: Matsushita
authors.forename: H
authors.initials: H
authors.lastname: Takahashi
authors.forename: K
authors.initials: K
authors.lastname: Kakishita
authors.forename: A
authors.initials: A
authors.lastname: Yamamoto
authors.forename: T
authors.initials: T
authors.lastname: Yoshizaki
authors.forename: T
authors.initials: T
authors.lastname: Terashima
authors.forename: F
authors.initials: F
authors.lastname: Murakami
authors.forename: T
authors.initials: T
authors.lastname: Itakura
authors.forename: H
authors.initials: H
authors.lastname: Okano
mesh_terms: D000818:Animals; D001369:Axons / Q000502:physiology; D002469:Cell Separation; D017690:Cell Transplantation; D002478:Cells, Cultured; D004195:Disease Models, Animal; D004298:Dopamine / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D008636:Mesencephalon / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D010300:Parkinson Disease; D051381:Rats; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics
chemical_list: D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine