Drs. Castelo-Branco, Andersson, and Minina contributed equally to this work.
Special Issue Research Article
Delayed dopaminergic neuron differentiation in Lrp6 mutant mice
Version of Record online: 30 SEP 2009
Copyright © 2009 Wiley-Liss, Inc.
Special Issue: Special Issue on WNT Signaling in Development and Disease
Volume 239, Issue 1, pages 211–221, January 2010
How to Cite
Castelo-Branco, G., Andersson, E. R., Minina, E., Sousa, K. M., Ribeiro, D., Kokubu, C., Imai, K., Prakash, N., Wurst, W. and Arenas, E. (2010), Delayed dopaminergic neuron differentiation in Lrp6 mutant mice. Dev. Dyn., 239: 211–221. doi: 10.1002/dvdy.22094
- Issue online: 15 DEC 2009
- Version of Record online: 30 SEP 2009
- Manuscript Accepted: 5 AUG 2009
- Swedish Research Council. Grant Numbers: VR2008 and 2811 and, DBRM
- Swedish Foundation for Strategic Research (INGVAR and CEDB), Norwegian Research Council, Karolinska Institutet, Michael J Fox Foundation, European Union (Eurostemcell)
- BMBF National Genome Research. Grant Number: FKZ 01GS08174
- Virtual Institute on Neurodegeneration and Ageing. Grant Number: VH-VI-252
- Initiative and Networking Fund in the framework of the Helmholtz Alliance of Systems Biology and of Mental Health in an Ageing Society. Grant Number: HA-215
- Bayerischer Forschungsverbund ‘ForNeuroCell’. Grant Number: F2-F2410-10c/20697
- European UnionmdDANEURODEV FP7-Health-2007-B-222999, EuTRACC LSHG-CT-2006-037445
- Deutsche Forschungsgemeinschaft (DFG)SFB 596, WU 164/3-2, WU 164/4-1
Additional Supporting Information may be found in the online version of this article.
|DVDY_22094_sm_SuppFig1.tif||764K||Supporting Information Figure 1. Temporal regulation of Lrp5 and Lrp6 expression during midbrain development. A,B: Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis revealed that Lrp5 (A) is expressed at constant levels in the developing ventral midbrain (VM), whereas Lrp6 (B) shows a peak of expression at embryonic day (E) 11.5. The dopaminergic (DA) neurogenic period in the rat ventral midbrain (VM) is highlighted with a box. (Error bars represent SEM of technical replicates as qRT-PCR was performed on pools of VMs). C: Detection of TH, Lrp5, and Lrp6 expression on sagittal sections of E11.5 wild-type mouse embryos by in situ hybridization. Lrp5 and Lrp6 are ubiquitously expressed in the brain, including the VM domain containing TH+ DA neurons. au, arbitrary units; te, telencephalon; ov, optic vesicle; di, diencephalon; mes, mesencephalon; hb, hindbrain.|
|DVDY_22094_sm_SuppFig2.tif||3956K||Supporting Information Figure 2. Lrp6−/− mice with exencephaly display normal ventral patterning, despite gross morphological dorsal defects. A,B: Shh (floor plate [FP] and basal plate [BP] marker), Lmx1b (FP and roof plate [RP] marker), and Wnt3a (RP marker) in situ hybridization of embryonic day (E) 9.5 (A) and E10.5 (B) wild-type (WT) and exencephalic Lrp6−/− mice revealed that patterning of the ventral domains is maintained, even though dorsal structures are grossly disrupted in the mutants. A: At E9.5, the ventricular invagination at the midline appears sharper and Shh and Lmx1b expression in the FP/BP might be broadened in Lrp6−/− mice. B: This difference is not seen in the mutants at E10.5.|
|DVDY_22094_sm_SuppFig3.tif||2320K||Supporting Information Figure 3. Proliferation, as assessed by BrdU (5-bromo-2-deoxyuridine) incorporation, is unchanged in Lrp6−/− mice. The proliferative capacity of embryonic day (E) 11.5 ventral midbrain (VM) precursors was not affected in Lrp6−/− VM as assessed by immunohistochemistry against BrdU in sagittal sections at E10.5, E11.5, E12.5, and E15.5.|
|DVDY_22094_sm_SuppFig4.tif||1597K||Supporting Information Figure 4. Cell death is not affected in Lrp6−/− mice. Cleaved caspase 3+ cells were virtually absent in wild-type and Lrp6−/− ventral midbrain (VM) at embryonic day (E) 11.5. All images were acquired with the same settings. Inset in each image is another region of the same brain showing positive staining.|
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