Fig 4.
Acer3 knockout decreases alkaline ceramidase activity on ULCC in the brain.
A. The transcription of a truncated coding sequence in a representative Acer3 knockout mouse. RNAs were isolated from the brains of Acer3+/+ or Acer3-/- mice and subjected to RT-PCR using a pair of primers encompassing the start codon and stop codon, respectively, of the Acer3 gene. Note that the Acer3-/- mouse has a smaller ORF of the Acer3 gene than an age-matched Acer3+/+ mouse. B. Reduction of alkaline ceramidase activity on NBD-C12-PHC in Acer3-/- mice. Note that Acer3-/- mice at either 6W or 8M of age show significant declines in ceramidase activity in both the cerebellar and cerebral brains compared to their WT littermates. C. Reduction of alkaline ceramidase activity on C18:1-ceramide in the whole brains of Acer3 knockout mice. Note that the brain alkaline ceramidase activity on this ceramide was substantially decreased in Acer3-/- mice compared to Acer3+/+ mice. Image in A represents result from 3 pairs of mice. Data in B and C represent mean values ± SD, n = 3.
Fig 9.
Acer3 knockout induces premature degeneration of PCs.
A and B. PC loss in Acer3 knockout mice at 8M of age. Immunostaining of cerebellar sagittal sections with antibody against calbindin D-28K, a PC marker (A). Red arrowheads indicate the regions where PCs were lost. Quantification of PCs (B). Images in A are the results from a representative mouse in each group. C. TUNEL assays for apoptosis in the cerebellum from Acer3+/+ and Acer3-/- mice. The cerebellar sections of Acer3+/+and Acer3-/- mice at 8M of age were co-stained with the TUNEL assay reagent (green fluorescence) and anti-calbindin D28K antibody (red fluorescence). The images in A and C are the results from a representative mouse in each group. The data in B represent mean values ± SD, n = 4.