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Cholesterol and ganglioside immunohistochemistry (IHC) of Npc1−/− and WT mice in the combination treatment study.

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posted on 11.09.2009 by Cristin D. Davidson, Nafeeza F. Ali, Matthew C. Micsenyi, Gloria Stephney, Sophie Renault, Kostantin Dobrenis, Daniel S. Ory, Marie T. Vanier, Steven U. Walkley

(A) Filipin labeling of unesterified cholesterol (seen as white areas in image) in the neocortex of age-matched untreated and treated Npc1−/− and WT mice (all mice between 75 and 81 days of age) revealed less cholesterol accumulation of treated Npc1−/− mice (second, third, and fourth panels) when compared to control Npc1−/− mice (fifth panel). WT mice do not exhibit cholesterol accumulation (first panel). Each panel here and in (B) and (C) shows layers II (top) through VI (bottom) of the cerebral cortex. (B) IHC of GM2 ganglioside (visualized as brown punctae within cells) was also characterized by reduced GM2 storage in all treated Npc1−/− mice. (C) IHC of GM3 ganglioside (again seen as brown punctae within cells) showed results similar to GM2. (D) Treated Npc1−/− mice had more remaining Purkinje cells (brown areas in cerebellar images) than did untreated Npc1−/− mice; however, treated mice still had Purkinje cell loss when compared to WT mice. Anti-calbindin antibody labels Purkinje cell bodies and dendritic arbors, while the Nissl counterstain (purple) labels all neuronal cell bodies. Images taken at 20X (A), 10X (B, C), and 2X (D); scale bars 20 µm (A), 50 µm (B, C), and 400 µm (D).

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