title: Human midbrain precursors activate the expected developmental genetic program and differentiate long-term to functional A9 dopamine neurons in vitro. Enhancement by Bcl-X(L).
abstract: Understanding the molecular programs of the generation of human dopaminergic neurons (DAn) from their ventral mesencephalic (VM) precursors is of key importance for basic studies, progress in cell therapy, drug screening and pharmacology in the context of Parkinson's disease. The nature of human DAn precursors in vitro is poorly understood, their properties unstable, and their availability highly limited. Here we present positive evidence that human VM precursors retaining their genuine properties and long-term capacity to generate A9 type Substantia nigra human DAn (hVM1 model cell line) can be propagated in culture. During a one month differentiation, these cells activate all key genes needed to progress from pro-neural and pro-dopaminergic precursors to mature and functional DAn. For the first time, we demonstrate that gene cascades are correctly activated during differentiation, resulting in the generation of mature DAn. These DAn have morphological and functional properties undistinguishable from those generated by VM primary neuronal cultures. In addition, we have found that the forced expression of Bcl-X(L) induces an increase in the expression of key developmental genes (MSX1, NGN2), maintenance of PITX3 expression temporal profile, and also enhances genes involved in DAn long-term function, maintenance and survival (EN1, LMX1B, NURR1 and PITX3). As a result, Bcl-X(L) anticipates and enhances DAn generation.
authors.affiliation: Departmento de Biología Molecular and Centro de Biología Molecular Severo Ochoa, Universidad Autónoma de Madrid-C.S.I.C, Cantoblanco, 28049-Madrid, Spain. egonzalez@cbm.uam.es
authors.forename: Emma G
authors.initials: EG
authors.lastname: Seiz
authors.forename: Milagros
authors.initials: M
authors.lastname: Ramos-Gómez
authors.forename: Elise T
authors.initials: ET
authors.lastname: Courtois
authors.forename: Jan
authors.initials: J
authors.lastname: Tønnesen
authors.forename: Merab
authors.initials: M
authors.lastname: Kokaia
authors.forename: Isabel
authors.initials: I
authors.lastname: Liste Noya
authors.forename: Alberto
authors.initials: A
authors.lastname: Martínez-Serrano
mesh_terms: D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D004298:Dopamine / Q000235:genetics / Q000378:metabolism; D059290:Dopaminergic Neurons / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008636:Mesencephalon / Q000166:cytology / Q000378:metabolism*; D016548:Prosencephalon / Q000378:metabolism; D013378:Substantia Nigra / Q000166:cytology / Q000254:growth & development / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D051020:bcl-X Protein / Q000235:genetics* / Q000378:metabolism*
chemical_list: C494811:BCL2L1 protein, human; D018398:Homeodomain Proteins; D014157:Transcription Factors; D051020:bcl-X Protein; C433629:homeobox protein PITX3; D004298:Dopamine