The Journal of Neuroscience, November 17, 2004, ():
Postnatal Neurogenesis and Gliogenesis in the Olfactory Bulb from NG2-Expressing Progenitors of the Subventricular Zone
J. Neurosci. Aguirre and Gallo
24: 10530
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplementary Figure S1. EGFP+ progenitors cells migrate caudally from the SVZ to the SCWM. (A-D) Reconstruction of a parasagittal section from a CNP-EGFP mouse brain 4 days after DiI injection into the LV (black arrow). A large percentage of double-labeled DiI+/EGFP+ (red/green) cells are found throughout the entire SCWM. In this region, a percentage of EGFP+ cells displayed a morphology typical of migratory cells (B). Boxes in A are shown at a higher magnification in panels B-D. Individual DiI+/EGFP+ labeled cells (white arrows; B1, C1 and D1) are further magnified in B2-4, C2-4 and D2-4. (E-F) A percentage of the DiI+/EGFP+ cells in the SCWM express NG2 (E4, blue) and O4 (F4, blue). Orthogonal reconstructions of confocal sections in the Z axes at the level indicated by the yellow lines are shown in panels E1 and E5, and F1 and F5. The individual cells selected for multi-marker illustration are indicated at the intersections of the yellow lines. Scale bar = 300µm (A), and 50µm (B-F).
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Supplementary Figure S2. Transplanted NG2+/EGFP+ cells migrate throughout the RMS and differentiate to OB interneurons. NG2+/EGFP+ cells were transplanted in the LV and tissues were analyzed at 4 days (B1-B2, and C1-C2) and 1 week (D-F). EGFP+ cells were visualized by fluorescence (480/520nm). (A) Diagram of a parasagittal section showing the migration pathway of EGFP+ cells at 4DAT. The green boxes in (A) are shown at higher magnification (B1-B2 and C1-C2), respectively. EGFP fluorescence was color converted to black and white to reveal the distribution of grafted EGFP+ cells. (D) The fluorescent dye PKH26 was used to pre-label FACS-purified NG2+/EGFP+ cells before transplantation (see also Aguirre et al., 2004). EGFP+/PKH26+ cells (D2 and D3, green/red, respectively) were observed in the RMShl. (E) EGFP+ cells expressed neuronal markers in the OB, including DC (E3, red) and Tuj (E4, blue). (F) EGFP+ cells in the OB expressed DC (F3, red) and Dlx (F4, blue). Boxes in B1 and C1 are shown at a higher magnification in B2 and C2, respectively. In all panels, cells indicated by arrows are shown at a higher magnification. Scale bar = 200µm (B1 and C1); 100µm (B2 and C2); 50µm (D-F).
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Supplementary Figure S3. NG2+/EYFP+ cells generate glial progeny in the SCWM after transplantation. All images were obtained 3 WAT. (A1-A3) A large number of grafted cells were found in the SCWM. Images in A1, A2 and A3 were obtained from the SCWM of three different mice after transplantation. (B1-B6) A large percentage of grafted EYFP+ cells expressed Ki67 (red). (C1-C5) A large percentage of grafted EYFP+ cells in the SCWM expressed mature oligodendrocyte markers, including CNP (C4, blue). (D1-D5) Graft-derived cells expressing GFAP (D3, red) were observed in the SCWM. Scale bar A1-A3 and B1-B3 = 100 µm; C1-D1 = 50 µm.
