title: Neurogenin2 identifies a transplantable dopamine neuron precursor in the developing ventral mesencephalon.
abstract: In neural transplantation studies, there is an interest in identifying and isolating mesencephalic dopamine (mesDA) neuron precursors that have the capacity to differentiate into fully mature mesDA neurons after transplantation. We report here that in the developing ventral mesencephalon (VM) the proneural gene Neurogenin2 (Ngn2) is expressed exclusively in the part of the ventricular zone that gives rise to the migrating mesDA neuroblasts, but not in the differentiated mesDA neurons. From other studies, we know that Ngn2 is involved in the generation of mesDA neurons and that the development of mesDA neurons is severely compromised in Ngn2-null mutant mice. We show here that cells isolated by FACS from the developing VM of Ngn2-GFP knock-in mice are capable of generating mesDA neurons, both in vitro and after transplantation to the striatum of neonatal rats. All mesDA neuron precursors, but not the serotonergic or GABAergic neuron precursors, are contained in the Ngn2-GFP-expressing population. Moreover, all glial cells were generated from cells contained in the GFP-negative cell fraction. The results show that surviving mesDA neurons in VM grafts are derived from early postmitotic, probably Nurr1-expressing precursors before they have acquired their fully differentiated neuronal phenotype. The Ngn2-GFP reporter construct used here thus provides a tool for the identification of mesDA neuron precursors in the VM and selective isolation of transplantable mesDA neuron precursors for transplantation.
authors.affiliation: Wallenberg Neuroscience Center, Department of Experimental Medical Science, and Lund Strategic Center for Stem Cell Biology and Cell Therapy, Lund University, BMC A11, SE-221 84 Lund, Sweden.
authors.forename: Lachlan H
authors.initials: LH
authors.lastname: Thompson
authors.forename: Elin
authors.initials: E
authors.lastname: Andersson
authors.forename: Josephine B
authors.initials: JB
authors.lastname: Jensen
authors.forename: Perrine
authors.initials: P
authors.lastname: Barraud
authors.forename: Francois
authors.initials: F
authors.lastname: Guillemot
authors.forename: Malin
authors.initials: M
authors.lastname: Parmar
authors.forename: Anders
authors.initials: A
authors.lastname: Björklund
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000378:metabolism; D000080924:Aldehyde Dehydrogenase 1 Family; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000172:deficiency / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D001973:Bromodeoxyuridine / Q000378:metabolism; D002452:Cell Count / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D004298:Dopamine / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018398:Homeodomain Proteins / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D017403:In Situ Hybridization / Q000379:methods; D007211:Indoles; D007527:Isoenzymes / Q000378:metabolism; D019394:Ki-67 Antigen / Q000378:metabolism; D008297:Male; D008636:Mesencephalon / Q000166:cytology* / Q000196:embryology / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D009419:Nerve Tissue Proteins / Q000172:deficiency / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D011247:Pregnancy; D050697:Retinal Dehydrogenase; D012701:Serotonin / Q000378:metabolism; D033581:Stem Cell Transplantation / Q000379:methods; D013234:Stem Cells / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
chemical_list: D000444:Aldehyde Dehydrogenase; D000080924:Aldehyde Dehydrogenase 1 Family; D051792:Basic Helix-Loop-Helix Proteins; D015415:Biomarkers; D001973:Bromodeoxyuridine; D002795:Choline O-Acetyltransferase; D004268:DNA-Binding Proteins; D004298:Dopamine; D005904:Glial Fibrillary Acidic Protein; D049452:Green Fluorescent Proteins; D018398:Homeodomain Proteins; D007211:Indoles; D007527:Isoenzymes; D019394:Ki-67 Antigen; D009419:Nerve Tissue Proteins; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D050697:Retinal Dehydrogenase; D012701:Serotonin; D014157:Transcription Factors; D014446:Tyrosine 3-Monooxygenase; D005680:gamma-Aminobutyric Acid; C102938:Neurog2 protein, mouse; C539883:Nr4a2 protein, mouse; C539884:Nr4a2 protein, rat; C007293:DAPI; C515028:ALDH1A1 protein, mouse; C523119:Aldh1a1 protein, rat