FoxP1 promotes midbrain identity in embryonic stem cell-derived dopamine neurons by regulating Pitx3.
The robust generation of midbrain dopamine neurons from embryonic stem cells and patient-specific induced pluripotent stem cells is a prospective tool for the development of new drugs and cell based therapies, and investigations into the aetiology of Parkinson's disease. To achieve this, it is crucial to identify the fate-determining regulatory factors that influence dopamine cell fate decision and the underlying molecular machinery. We identified FoxP1 as a novel marker for midbrain dopamine neurons. Enforced expression of FoxP1 in embryonic stem cells actuates the expression of Pitx3, a homeobox protein that is exclusively expressed in midbrain dopaminergic neurons and is required for their differentiation and survival during development and from embryonic stem cells in vitro. We show that FoxP1 can be recruited to the Pitx3 locus in embryonic stem cells and regulate Pitx3 promoter activity in a dual-luciferase assay. This transcriptional regulation of Pitx3 by FoxP1 depends on the presence of two high affinity binding sites in the distal Pitx3 promoter, through which FoxP1 directly binds as demonstrated by chromatin immunoprecipitation and electrophoretic mobility shift assay. Thus, this study demonstrates for the first time a transcription regulatory role for FoxP1 on the Pitx3 gene in mammalian stem cells.
Konstantoulas
Constantine James
CJ
MRC Clinical Sciences Centre, Faculty of Medicine, Imperial College, London, UK. constantine.konstantoulas@imperial.ac.uk
Parmar
Malin
M
Li
Meng
M
D000818:Animals; D000831:Animals, Newborn; D001665:Binding Sites / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D004298:Dopamine / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000502:physiology; D051381:Rats; D050436:Regulatory Elements, Transcriptional / Q000502:physiology; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D033581:Stem Cell Transplantation / Q000379:methods; D013378:Substantia Nigra / Q000166:cytology / Q000196:embryology*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D051858:Forkhead Transcription Factors; C432917:Foxp1 protein, mouse; D018398:Homeodomain Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; C433629:homeobox protein PITX3; D004298:Dopamine