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

Autor:	Antonio Jiménez-Beristain, Jiexin Zhao, Carmel M. O’Brien, Carmen De Felipe, Meng Li, Elena V. Semina, Joaquim Vives, Suling Zhao, Sarah L. Maxwell, Eva Kuehner
Přispěvatelé:	Medical Research Council (UK), Stem Cell Network, Consejo Nacional de Ciencia y Tecnología (México)
Rok vydání:	2004
Předmět:	Male Dopamine Green Fluorescent Proteins Mice Transgenic homologous recombination Substantia nigra Pitx3 Biology Green fluorescent protein Mice Mesencephalon Pregnancy Neurotrophic factors medicine Animals Cells Cultured Homeodomain Proteins Neurons Dopaminergic neuron Stem Cells General Neuroscience green fluorescent protein (GFP) Gene Expression Regulation Developmental homeobox transcription factor Embryo Mammalian Embryonic stem cell Molecular biology eye diseases Neural stem cell Mice Inbred C57BL stem cell Ventral tegmental area Luminescent Proteins medicine.anatomical_structure nervous system Neuron differentiation Female Stem cell Transcription Factors
Zdroj:	European Journal of Neuroscience. 19:1133-1140
ISSN:	1460-9568 0953-816X
DOI:	10.1111/j.1460-9568.2004.03206.x
Popis:	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. This work is supported by grants from the UK Medical Research Council (MRC) and an MRC–Stem Cell Science UK Ltd Collaborative PhD studentship (S.M.), and a CONCYTEA Mexico Studentship (A.J.-B.).
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b5eb5ce1874fbe9dcfdbb2557c6a1788 https://doi.org/10.1111/j.1460-9568.2004.03206.x Zobrazit plný text záznamu Plný text ve formátu PDF Plný text ve formátu HTML
Nepřihlášeným uživatelům se plný text nezobrazuje	K zobrazení výsledku je třeba se přihlásit.