Differentiation of embryonic stem cell-derived dopaminergic neurons is enhanced by survival-promoting factors.
Here, we describe the generation of viable and dopamine-producing neurons derived from pluripotent mouse embryonic stem cells. Neurotrophic factors in combination with survival-promoting factors, such as interleukin-1beta, glial cell line-derived neurotrophic factor, neurturin, transforming growth factor-beta(3) and dibutyryl-cyclic AMP, significantly enhanced Nurr1 and tyrosine hydroxylase (TH) mRNA levels, whereas En-1, mash-1 and dopamine-2-receptor mRNA levels were not upregulated. In parallel, mRNA levels of the anti-apoptotic gene bcl-2 were found to be upregulated at terminal stages. Double immunofluorescence analysis revealed increased numbers of TH- and dopamine transporter-, but not gamma-aminobutyric acid- and serotonin-positive neurons in relation to synaptophysin-labeled cells by survival-promoting factors. Moreover, high-performance liquid chromatography analysis showed detectable levels of intracellular dopamine. We conclude that survival-promoting factors enhance differentiation, survival and maintenance of dopaminergic neurons derived from embryonic stem cells.
Rolletschek
A
A
'In vitro Differentiation' Group, Institute of Plant Genetics and Crop Plant Research, Corrensstrasse 3, D-06466, Gatersleben, Germany.
Chang
H
H
Guan
K
K
Czyz
J
J
Meyer
M
M
Wobus
A M
AM
D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis; D051792:Basic Helix-Loop-Helix Proteins; D003994:Bucladesine / Q000378:metabolism; D002454:Cell Differentiation; D002470:Cell Survival; D002851:Chromatography, High Pressure Liquid; D004268:DNA-Binding Proteins / Q000096:biosynthesis; D004298:Dopamine / Q000096:biosynthesis* / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D018398:Homeodomain Proteins / Q000096:biosynthesis; D007150:Immunohistochemistry; D051379:Mice; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012701:Serotonin / Q000096:biosynthesis; D013997:Time Factors; D014157:Transcription Factors / Q000096:biosynthesis / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism; D053782:Transforming Growth Factor beta3; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism; D015854:Up-Regulation; D005680:gamma-Aminobutyric Acid / Q000096:biosynthesis
D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; D051792:Basic Helix-Loop-Helix Proteins; D003994:Bucladesine; D004268:DNA-Binding Proteins; D004298:Dopamine; D018398:Homeodomain Proteins; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D012701:Serotonin; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; D053782:Transforming Growth Factor beta3; D014446:Tyrosine 3-Monooxygenase; D005680:gamma-Aminobutyric Acid; C499317:Ascl1 protein, mouse; C080330:En1 protein, mouse; C539883:Nr4a2 protein, mouse; C509045:Tgfb3 protein, mouse