Generation of embryonic stem cells and transgenic mice expressing green fluorescence protein in midbrain dopaminergic neurons

Zhao, Suling, Maxwell, Sarah, Jimenez-Beristain, Antonio, Vives, Joaquim, Kuehner, Eva, Zhao, Jiexin, O'Brien, Carmel, de Felipe, Carmen, Semina, Elena and Li, Meng ORCID: https://orcid.org/0000-0002-4803-4643 2004. Generation of embryonic stem cells and transgenic mice expressing green fluorescence protein in midbrain dopaminergic neurons. European Journal of Neuroscience 19 (5) , pp. 1133-1140. 10.1111/j.1460-9568.2004.03206.x

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Abstract

We have generated embryonic stem (ES) cells and transgenic mice with green fluorescent protein (GFP) inserted into the Pitx3 locus via homologous recombination. In the central nervous system, Pitx3-directed GFP was visualized in dopaminergic (DA) neurons in the substantia nigra and ventral tegmental area. Live primary DA neurons can be isolated by fluorescence-activated cell sorting from these transgenic mouse embryos. In culture, Pitx3–GFP is coexpressed in a proportion of ES-derived DA neurons. Furthermore, ES cell-derived Pitx3–GFP expressing DA neurons responded to neurotrophic factors and were sensitive to DA-specific neurotoxin N-4-methyl-1, 2, 3, 6-tetrahydropyridine. We anticipate that the Pitx3–GFP ES cells could be used as a powerful model system for functional identification of molecules governing mDA neuron differentiation and for preclinical research including pharmaceutical drug screening and transplantation. The Pitx3 knock-in mice, on the other hand, could be used for purifying primary neurons for molecular studies associated with the midbrain-specific DA phenotype at a level not previously feasible. These mice would also provide a useful tool to study DA fate determination from embryo- or adult-derived neural stem cells.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Biosciences Medicine
Subjects:	Q Science > Q Science (General)
Uncontrolled Keywords:	Dopaminergic neuron; green fluorescent protein (GFP); homologous recombination; homeobox transcription factor; Pitx3; stem cell.
Publisher:	Wiley
ISSN:	0953-816X
Last Modified:	27 Oct 2022 08:41
URI:	https://orca.cardiff.ac.uk/id/eprint/63104

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