Open Access Highly Accessed Research article

Temporal-spatial changes in Sonic Hedgehog expression and signaling reveal different potentials of ventral mesencephalic progenitors to populate distinct ventral midbrain nuclei

Sandra Blaess12*, Gabriela O Bodea2, Anna Kabanova2, Soline Chanet3, Emilie Mugniery4, Amin Derouiche56, Daniel Stephen1 and Alexandra L Joyner1

Author Affiliations

1 Developmental Biology Program, Sloan-Kettering Institute, 1275 York Avenue, New York, NY 10021, USA

2 Institute of Reconstructive Neurobiology, Life and Brain Center, University of Bonn, Sigmund-Freud-Str. 25, 53127 Bonn, Germany

3 Institut Pasteur, GDD Unit, CNRS URA 2578, 25 rue du Dr Roux, 75015 Paris, France

4 4Hôpital Necker - Enfants Malades, 149 rue de Sèvres - 75743 Paris cedex 15, France

5 Institute of Cellular Neurosciences, University of Bonn, Sigmund-Freud-Str. 25, 53105 Bonn, Germany

6 Institute for Anatomy II, Goethe-Universität Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt am Main, Germany

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Neural Development 2011, 6:29  doi:10.1186/1749-8104-6-29

Published: 20 June 2011

Additional files

Additional file 1:

Comparison of the extent of labeling using the ShhCreERneoand ShhCreERalleles with R26lzreporter mice. Coronal sections of E18.5 brains were labeled with X-gal staining and counterstained with Fast Red. TM (4 mg) was given at E10.5. (A-F) Note that only few cells are labeled in the midbrain of ShhCreERneo/+R26lz/+ mice (A-C), while many cells are labeled in the midbrain of ShhCreER/+R26lz/+ mice (D-F). Pictures in the upper right corner are higher magnifications of the area indicated in the black box. Several images were taken for each area shown and stitched together using the Zeiss Mosaix software.

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Additional file 2:

Initial domains of cells marked with Shh- or Gli1-GIFM in comparison with Lmx1a. (A-H') TM was administered at the indicated time points and marked cells were analyzed at E12.5 on anterior and posterior coronal midbrain sections with EYFP (red) and Lmx1a (green) immunostaining. Note that on anterior sections there is no or only little overlap of Lmx1a with cells fate-mapped with Shh-GIFM at E11.5 or Gli1-GIFM at E9.5. Scale bars: (A-H) 100 μm; (B',D',F',H') 50 μm. (I-L) Number of fate-mapped cells overlapping with Lmx1a (I,K) or TH (J,L). Cells were counted on one anterior (ant), one intermediate (int) and one posterior (pos) coronal section for each E12.5 embryo (or at E11.5 for Shh-GIFM TM8.5). For one time point and one section level, each data point represents cell numbers from one embryo. Note that overlap of fate-mapped cells is much higher with Lmx1a than with TH, since Lmx1a is expressed in DA precursors and differentiated DA neurons and differentiation of DA neurons is not complete by E12.5. The trends observed at E12.5 correlate with the results of the analysis at E18.5 and in the adult brain sections: the highest contribution to DA neurons is observed with Shh-GIFM at E9.5 (I,J); cells marked with Shh-GIFM TM8.5 contribute preferentially to anterior DA neurons (J); cells marked with Shh-GIFM TM11.5 contribute preferentially to intermediate and posterior DA neurons (J); Gli1-GIFM results in less labeling than Shh-GIFM (compare (I,J) with (K,L)).

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Additional file 3:

Markers used to determine the location of Shh- and Gli1-derived precursors in relation to known ventral midbrain expression domains.

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Additional file 4:

Contribution of fate-mapped cells to different regions of DA neurons along the rostral-caudal axis of the prenatal midbrain and to different subsets of DA neurons in the adult midbrain. (A,B) Number of Shh-derived (A) or Gli1-derived (B) cells contributing to different rostral-caudal regions of DA neurons at E18.5. For each animal (n ≥ 3), β-gal- and TH-co-expressing cells were counted in four regions along the rostral-caudal axis of the ventral midbrain as indicated in Figure 5B and normalized for the number of sections counted for each region. (C,D) Number of Shh-derived (C) or Gli1-derived (D) cells contributing to different subsets of DA neurons in the adult brain. For each animal (n ≥ 3), β-gal- and TH-co-expressing cells were counted in the SN, dlVTA and vmVTA as indicated in Figure 6L and normalized for the number of sections counted for each region. (E,F) Relative distribution of DA neurons in the SN, dlVTA and vmVTA (black bars) compared to the relative contribution of Shh-GIFM marked cells to the three areas. For a clearer representation, the data were split into two diagrams. The data for the fate-mapped cells (TM8.5, TM9.5, TM10.5, TM11.5) are also shown in Figure 6I. Error bars indicate standard deviation. Significance (*P < 0.05; **P < 0.01; ***P < 0.001) was determined by ANOVA and LSD post-hoc analysis (A-D) or Student's t-test (E,F).

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