title: Retinoic acid-dependent and -independent gene-regulatory pathways of Pitx3 in meso-diencephalic dopaminergic neurons.
abstract: Development of meso-diencephalic dopamine (mdDA) neurons requires the combined actions of the orphan nuclear receptor Nurr1 and the paired-like homeobox transcription factor Pitx3. Whereas all mdDA neurons require Nurr1 for expression of Th and survival, dependence on Pitx3 is displayed only by the mdDA subpopulation that will form the substantia nigra (SNc). Previously, we have demonstrated that Pitx3(-/-) embryos lack the expression of the retinoic acid (RA)-generating enzyme Ahd2, which is normally selectively expressed in the Pitx3-dependent DA neurons of the SNc. Restoring RA signaling in Pitx3(-/-) embryos revealed a selective dependence of SNc neurons on the presence of RA for differentiation into Th-positive neurons and maintenance throughout embryonic development. Whereas these data are suggestive of an important developmental role for RA in neurons of the SNc, it remained unclear whether other Nurr1 and Pitx3 target genes depend on RA signaling in a manner similar to Th. In the search for genes that were affected in Pitx3-deficient mdDA neurons and restored upon embryonic RA treatment, we provide evidence that Delta-like 1, D2R (Drd2) and Th are regulated by Pitx3 and RA signaling, which influences the mdDA terminal differentiated phenotype. Furthermore, we show that regulation of Ahd2-mediated RA signaling represents only one aspect of the Pitx3 downstream cascade, as Vmat2, Dat, Ahd2 (Aldh1a1), En1, En2 and Cck were unaffected by RA treatment and are (subset) specifically modulated by Pitx3. In conclusion, our data reveal several RA-dependent and -independent aspects of the Pitx3-regulated gene cascade, suggesting that Pitx3 acts on multiple levels in the molecular subset-specification of mdDA neurons.
authors: Rudolf Magnus Institute, Department of Neuroscience and Pharmacology, UMC Utrecht, Utrecht, The Netherlands.
authors: Frank M J
authors: FM
authors: Jacobs
authors: Jesse V
authors: JV
authors: Veenvliet
authors: Wadia H
authors: WH
authors: Almirza
authors: Elisa J
authors: EJ
authors: Hoekstra
authors: Lars
authors: L
authors: von Oerthel
authors: Annemarie J A
authors: AJ
authors: van der Linden
authors: Roel
authors: R
authors: Neijts
authors: Marian Groot
authors: MG
authors: Koerkamp
authors: Dik
authors: D
authors: van Leenen
authors: Frank C P
authors: FC
authors: Holstege
authors: J Peter H
authors: JP
authors: Burbach
authors: Marten P
authors: MP
authors: Smidt
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D002135:Calcium-Binding Proteins; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D004027:Diencephalon / Q000166:cytology* / Q000196:embryology*; D005260:Female; D005455:Fluorescent Antibody Technique; D053263:Gene Regulatory Networks / Q000187:drug effects* / Q000502:physiology; D005838:Genotype; D018398:Homeodomain Proteins / Q000378:metabolism*; D017403:In Situ Hybridization; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D046228:Microarray Analysis; D009474:Neurons / Q000378:metabolism* / Q000502:physiology; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2 / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D017448:Receptors, Dopamine D2 / Q000378:metabolism; D014157:Transcription Factors / Q000172:deficiency / Q000378:metabolism*; D014212:Tretinoin / Q000494:pharmacology*
chemical_list: D002135:Calcium-Binding Proteins; C512925:Dlk1 protein, mouse; D018398:Homeodomain Proteins; D036341:Intercellular Signaling Peptides and Proteins; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D017448:Receptors, Dopamine D2; D014157:Transcription Factors; C433629:homeobox protein PITX3; D014212:Tretinoin