Journal: The Journal of Neuroscience

Article Title: Specification of Optic Nerve Oligodendrocyte Precursors by Retinal Ganglion Cell Axons

doi: 10.1523/JNEUROSCI.0855-06.2006

Figure Lengend Snippet: Retinal induction of OPCs is dependent on hedgehog signaling. A–F, Stage 24 (E4) retinal explant conditioned medium induces OPCs in stage 24 (E4) dorsal spinal cord cells after 4 d. A, B, Control cultures containing few OPCs. C, D, Retinal CM induces significant numbers of OPCs. E, F, This induction is inhibited by treatment with cyclopamine. G, Quantification of O4+ cells in cultures treated with Shh function-blocking mAb 5E1 shows that the induction is inhibited in a dose-dependent manner. Error bars represent SD (3 different preparations). H, I, In cocultures of retinal and dorsal spinal cord explants, the induction of OPCs is reduced significantly by cyclopamine treatment. J, Quantification of O4+ cells in control cocultures and cocultures treated with cyclopamine. K, L, Likewise, the inductive capacity of retinal neurites in the Campenot chambers is blocked by the addition of cyclopamine to the lateral chamber. M, Quantification of O4+ cells in Campenot chambers. Scale bars (in D, K), 50 μm.

Article Snippet: 5E1 antibody (Developmental Studies Hybridoma Bank) and anti-NRG antibody were added to the culture.

Techniques: Blocking Assay

Journal: The Journal of Neuroscience

Article Title: Specification of Optic Nerve Oligodendrocyte Precursors by Retinal Ganglion Cell Axons

doi: 10.1523/JNEUROSCI.0855-06.2006

Figure Lengend Snippet: Appearance of OPCs at the midline of the third ventricle is blocked by cyclopamine treatment, and localized Shh expression is absent in eyeless animals. A, B, Exposure to cyclopamine between stage 21 (E3) and stage 29 (E6) inhibited the normal development of OPCs lining the third ventricle. C–H, Explants derived from the third ventricle floor developed reduced numbers of O4+ cells after cyclopamine exposure. Shown are control explants (C, D), explants from cyclopamine-injected animals (E, F), and explants exposed to cyclopamine in vitro (G, H). I, Quantification of OPCs in explants from animals exposed to cyclopamine. J, Quantification of OPCs in explants treated with cyclopamine after removal from the naive animals. The reduction in OPC number was greater in explants from animals exposed to cyclopamine (I) than in explants treated with cyclopamine after removal (J), suggesting that OPC induction occurred before explant establishment. K, Shh is detectable by mAb 5E1 staining in the chiasmal region (arrow) of the optic nerve only in the presence of optic nerve axons. L, In eyeless animals, the labeling is lost, although it is retained in lateral aspects of the third ventricle (arrowhead). M, Expression of patched is detected at the floor of the third ventricle. N, O, In wild-type embryos at stage 27 (E5) and stage 31 (E7) Shh mRNA is not expressed at the floor of the third ventricle adjacent to the optic chiasm, although it is expressed laterally (on, optic nerve). P, At stage 31 (E7) in eyeless embryos the pattern of Shh mRNA expression was not altered significantly. Scale bars: A, B, M, O, P, 100 μm; C–H, K, L, N, 50 μm.

Article Snippet: 5E1 antibody (Developmental Studies Hybridoma Bank) and anti-NRG antibody were added to the culture.

Techniques: Expressing, Derivative Assay, Injection, In Vitro, Staining, Labeling