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. 2011 Jan 12;31(2):480–491. doi: 10.1523/JNEUROSCI.4698-10.2011

Figure 8.

Figure 8.

Migratory neurons in nasal region are born before VNO sensory neurons. A, Experimental paradigm: pregnant mice were injected with a single BrdU injection (50 μg/g) at E10.5, and the distribution of BrdU-positive neurons in the VNO anlage and in the nasal area was analyzed 24 h (E11.5) to 96 h (E14.5) after injection. The distribution of BrdU-labeled neurons between VNO and nasal area is schematically represented. B, E11.5 Hu (green)/BrdU (red) double labeling, 24 h after BrdU injection. Hu/BrdU double-positive neurons were distributed within (arrow) and migrating out of (arrowhead) the VNO. C, E12.5 Hu (green)/BrdU (red) double labeling 48 h after BrdU injection. Few Hu-positive and BrdU heavy labeled cells (arrow) were found within the VNO but were detected emerging from the VNO (arrowhead). D, E, E13.5, 72 h after BrdU injection, Hu/BrdU cells (D) and GnRH-1/BrdU-positive cells (E) were detected in the nasal area (NA). F, Second experimental paradigm: pregnant mice were injected with a single BrdU injection (50 μg/g) at the indicated embryonic stage and the BrdU incorporation in migratory neurons and in non-migratory VNO neurons was analyzed at E14.5. Schematic shows BrdU incorporation in migratory neurons declined as VNO neurogenesis increased. Sections of VNO immunolabeled for BrdU after single BrdU pulse at E10.5 (G, H), E11.5 (I), and E12.5 (J). G, Double labeling for GnRH-1 (brown) and BrdU (black) at E14.5 after BrdU pulse at E10.5 showed migratory cells and double-labeled BrdU/GnRH-1 (arrowheads) in the nasal area (NA), whereas only a few heavy BrdU-labeled cells (arrow) were detected in the VNO. Inset, Double-labeled BrdU/GnRH-1 neuron. H, I, Few heavy labeled BrdU-positive cells (black) were detected (arrow) in the VNO after BrdU injections at E10.5 and E11.5. J, Heavy labeled cells were detected in the VNO after BrdU injection at E12.5, and the pattern of these BrdU labeled cells was similar to postmitotic neurons stained for Hu (brown, K) at E14.5 within the VNO (arrow), although Hu-positive cells were still detected migrating out of (arrowhead) the VNO. Red dashed line, VNO. Scale bars: A, 50 μm; C, inset in G, 25 μm; G, 150 μm.