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. 2016 May 18;7:11628. doi: 10.1038/ncomms11628

Figure 5. Properties and progeny of postnatal fourth VZ progenitor cells.

Figure 5

(a) Wild-type mice received BrdU in a single dose 100 min before perfusion. BrdU+ nuclei in postnatal 4th VZ are surrounded by fibres of GFAP and vimentin. Scale bar, 20 μm. (b) Co-staining for M-phase marker phospho-histone-H3 (pH3) and phospho-ser55-Vimentin (p-Vimentin) reveals radial glial-like morphology of 4th VZ progenitors. Scale bar, 20 μm. (c–i) Viral fate mapping of postnatal 4th VZ progenitors. Ai14 or Ai14;ALDH1L1:GFP pups received Adeno-Cre viral injection in lateral ventricle at age P1, and were perfused at P4 (100 min after a single dose of EdU) or at P24. (d) Co-staining of P4 tissue sections for EdU, GFAP, and TdTomato reveals proliferative TdT+ nuclei in the VZ of the 4th ventricle. Scale bar, 20 μm. (e) Cre-recombined cells are visible in olfactory bulb, confirming successful targeting of forebrain V-SVZ progenitors; along the surface of the entire ventricular system, including pontine 4th ventricle; and along the ventral surface of the brain, suggesting viral outflow to meningeal space via cisterna magna. Scale bar, 1 mm. (f) Enlarged view of 4th VZ and pontine tegmentum region indicated in (e), revealing parenchymal TdTomato (TdT)+ cells. Scale bar, 100 μm. (g) Further enlargment of pontine tegmentum as indicated in (f), showing co-immunostaining for TdTomato (Red) and lineage-specific markers (Green: left, Sox10; right, ALDH1L1:GFP). Scale bar, 50 μm. (h) Enlarged view of basis pontis region indicated in (e), revealing parenchymal TdT+ cells. Scale bar, 100 μm. (i) Further enlargment of basis pontis as indicated in h, showing immunostaining for TdT and lack of colocalization with either Sox10 (left) or ALDH1L1:GFP (right). Many TdT+ cells are found along structures morphologically consistent with blood vessels. Scale bar, 50 μm.