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Figure 7 | Neural Development

Figure 7

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

Figure 7

Cells labeled with Gli1-GIFM at different time points have a changing potential to contribute to subpopulations of DA neurons. (A-D) Representative schematics of immunostained sections (P14 to P60) labeled with Gli1-GIFM at E9.5. Red dots, TH-positive fate-mapped cells; yellow crosses, cells with astrocytic morphology. Rostral, Bregma -2.92; caudal, Bregma -3.40 [20]. If, interfascicular nucleus; Pn, paranigral nucleus; Snc, substantia nigra pars compacta; Snl, substantia nigra lateralis; Vta, VTA. (E-I') Immunofluorescent staining for DA neurons (TH, green) and fate-mapped cells (Gli1-GIFM at E9.5; β-gal, red) on coronal midbrain sections. The areas in (F,F'), (H,H') and (I) are indicated in (E) and (G). Big arrows indicate TH- and β-gal-positive cells; arrowheads indicate fate-mapped cells with astrocytic morphology; small arrows indicate β-gal-positive, TH-negative cells with neuronal morphology. (I,I') Z-stacks of optical sections taken with a Zeiss Apotome. (I') Area indicated with dashed box in (I). Scale bars: (E,G) 200 μm; (F,H) 40 μm; (I,I') 20 μm. (J) Relative contribution of cells marked with Gli1-GIFM between E7.5 and E9.5 to the SN, dlVTA and vmVTA; see schematic in Figure 6L. For each animal (n ≥ 3), TH-positive fate-mapped cells were counted in the three indicated areas and normalized for the combined number of overlapping cells counted in these areas (in percent). Error bars indicate standard deviation. Significance (*P < 0.05; **P < 0.01) was determined by ANOVA and LSD post-hoc analysis. (K) Fate mapping strategy. (L) Relative contribution of fate-mapped cells to different rostral-caudal midbrain regions at E18.5. For each animal (n ≥ 3), β-gal- and TH-co-expressing cells were counted in four rostral-caudal midbrain regions (see (M)) and normalized for the combined number of cells counted in the four regions (in percent). Error bars indicate standard deviation. Significance (*P < 0.05; **P < 0.01) was determined by ANOVA and LSD post-hoc analysis. (M) Rostral-caudal areas used to quantify the contribution of fate-mapped cells to DA neurons at E18.5. RRF, retrorubral field.

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