Visualization, direct isolation, and transplantation of midbrain dopaminergic neurons

Kazunobu Sawamoto, Naoyuki Nakao, Kazuto Kobayashi, Natsuki Matsushita, Hiroshi Takahashi, Koji Kakishita, Atsuyo Yamamoto, Takahito Yoshizaki, Toshio Terashima, Fujio Murakami, Toru Itakura, Hideyuki Okano

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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.

Original languageEnglish
Pages (from-to)6423-6428
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume98
Issue number11
DOIs
Publication statusPublished - 2001 May 22

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Cite this

Sawamoto, K., Nakao, N., Kobayashi, K., Matsushita, N., Takahashi, H., Kakishita, K., Yamamoto, A., Yoshizaki, T., Terashima, T., Murakami, F., Itakura, T., & Okano, H. (2001). Visualization, direct isolation, and transplantation of midbrain dopaminergic neurons. Proceedings of the National Academy of Sciences of the United States of America, 98(11), 6423-6428. https://doi.org/10.1073/pnas.111152398