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. 2016 Dec 20;133(3):463–483. doi: 10.1007/s00401-016-1658-6

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© The Author(s) 2016

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

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Fig. 5 — Free-choice alcohol drinking re-establishes sphingomyelin homeostasis in the nucleus accumbens (Nac) of mice over-expressing acid sphingomyelinase (tgASM). The mass spectrum generated in a MALDI-MS imaging contains hundreds of ion signals associated with each individual pixel. The MALDI images were acquired with x–y-raster widths of 30 µm for coronal sections. Slice MALDI mass spectrograms for the three most abundant sphingomyelin (SM) species in the neuropil of the Nac of water or 16 vol% alcohol (EtOH) drinking tgASM or wild type (WT) mice with regular consumption and three withdrawal and reinstatement episodes. The Nac was analyzed as a target area of interest. The error bars show a mean of nine samples/genotype-treatment (±SEM) for four animals. The data are shown using a rainbow scale, normalized against the total ion count. a The coronal tissue sections show SM 18:1 18:0 highly enriched in the Nac. b The abundance of SM 18:1 18:0 is largely reduced in tgASM mice (ANOVA, factor genotype: F _1,32 = 240.793, p < 0.001). Alcohol drinking reduces levels of SM 18:1 18:0 in WT mice, but enhances it in tgASM mice (ANOVA, genotype × treatment interaction: F _1,32 = 196.600, p < 0.001). c The coronal tissue sections show SM 18:1 18:1 that is highly enriched in the Nac and piriform cortex. d The abundance of SM 18:1 18:1 is largely reduced in tgASM mice (ANOVA, factor genotype: F _1,32 = 165.064, p < 0.001). Alcohol drinking reduces levels of SM 18:1 18:1 in WT mice but enhances it in tgASM mice (ANOVA, genotype × treatment interaction: F _1,32 = 161.448, p < 0.001). e The coronal tissue sections show SM 18:1 20:0 that is highly enriched in the somatosensory and motor cortex and in dorsolateral striatum. f The abundance of SM 18:1 20:0 is reduced in tgASM mice (ANOVA, factor genotype: F _1,32 = 174.123, p < 0.001). Alcohol drinking reduces levels of SM 18:1 20:0 in WT mice but enhances it in tgASM mice (ANOVA, factor treatment: F _1,32 = 7.292, p < 0.011; genotype × treatment interaction: F _1,32 = 163.235, p < 0.001). g Mass spectrogram for a single slice at the Nac level with SM target masses. h Anatomical analysis of the slice at the level of the Nac (ac commisura anterior, CiC cingular cortex, CoCa corpus callosum, CPu caudate putamen, DB diagonal band of Broca, lv ventricle, InC insular cortex, MoC motor cortex, Nac/c Nac core, Nac/s Nac shell, PiC piriform cortex, SoSeC somatosensory cortex). For an overview on white matter and the liquor associated SM species, see Suppl. Figure 6, 7 (***p < 0.001; ^# p < 0.001 vs. WT-water)