Journal: Neurobiology of Disease

Article Title: A marked paucity of granule cells in the developing cerebellum of the Npc1 −/− mouse is corrected by a single injection of hydroxypropyl-β-cyclodextrin

doi: 10.1016/j.nbd.2014.06.012

Figure Lengend Snippet: Npc1 −/− mice display reduced granule neuron density in the EGL that is rescued by the HPBCD treatment. (A,C) Cerebellar sections of PN11, PN14 Npc1 +/+ , and Npc1 −/− mice, either untreated or treated with a single injection of HPBCD at PN7, were stained with Hoechst 33258. Representative sections are shown in the figure. Higher magnification fields of EGL base or crown of lobules II and X are shown on the right of low magnification fields. No major differences were detected at PN11. At PN14 however, the EGL of both anterior and posterior lobules of Npc1 −/− mice was thinner than that of age-matched Npc1 +/+ , while HPBCD treatment fully rescued this phenotype. (B,D) Histograms represent granule neuron densities (mean ± SEM of all sections examined; 4–5 mice/group; 3–4 sections/mouse) determined in 100 μm 2 regions of the crowns of the anterior (I–V) and posterior (VI–X) lobules of Npc1 +/+ (empty bars), Npc1 −/− (full bars) and HPBCD-treated Npc1 −/− (dashed bars) mice. Asterisks indicate statistically significant differences (* p < 0.05; ** p < 0.005). Scale bars indicate 250 μm (panels) and 50 μm (inserts).

Article Snippet: To determine the number of GN cells, sections from PN11 and PN14 mice were stained with 0.5 μg/mL Hoechst (Hoechst-33258, Invitrogen, Milan, Italy) for 4 min. To assess GN proliferation, brains were dissected from Npc1 +/+ and Npc1 −/− mice, either treated or untreated with HBCD and that had received a single BrdU injection at PN10, PN12 and PN14, and processed as described above, obtaining parasagittal sections to be stained with anti-BrdU antibodies.

Techniques: Injection, Staining