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

Generation of embryonic stem cells and transgenic mice expressing green fluorescence protein in... 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png European Journal of Neuroscience Wiley

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

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Publisher
Wiley
Copyright
Copyright © 2004 Wiley Subscription Services, Inc., A Wiley Company
ISSN
0953-816X
eISSN
1460-9568
DOI
10.1111/j.1460-9568.2004.03206.x
Publisher site
See Article on Publisher Site

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.

Journal

European Journal of NeuroscienceWiley

Published: Mar 1, 2004

References

  • CNS expression pattern of Lmx1b and coexpression with ptx genes suggest functional cooperativity in the development of forebrain motor control systems
    Asbreuk, Asbreuk; Vogelaar, Vogelaar; Hellemons, Hellemons; Smidt, Smidt; Burbach, Burbach
  • Glial cell line‐derived neurotrophic factor protects dopaminergic neurons from 6‐hydroxydopamine toxicity in vitro
    Eggert, Eggert; Schlegel, Schlegel; Oertel, Oertel; Wurz, Wurz; Krieg, Krieg; Vedder, Vedder
  • Neuroprotective and neurorestorative properties of GDNF
    Gash, Gash; Zhang, Zhang; Gerhardt, Gerhardt
  • Generation of purified neural precursors from embryonic stem cells by lineage selection
    Li, Li; Pevny, Pevny; Lovell‐Badge, Lovell‐Badge; Smith, Smith
  • A double‐deletion mutation in the Pitx3 gene causes arrested lens development in aphakia mice
    Rieger, Rieger; Reichenberger, Reichenberger; McLean, McLean; Sidow, Sidow; Olsen, Olsen
  • Isolation and characterization of a novel human paired‐like homeodomain‐ containing transcription factor gene, VSX1, expressed in ocular tissues
    Semina, Semina; Mintz‐Hittner, Mintz‐Hittner; Murray, Murray
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  • Culture and differentiation of embryonic stem cell
    Smith, Smith

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