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Fig. 2 | Molecular Neurodegeneration

Fig. 2

From: Loss of Munc18-1 long splice variant in GABAergic terminals is associated with cognitive decline and increased risk of dementia in a community sample

Fig. 2

Immunohistochemical characterization of M18 splice variants in rat brain. (a) Photomicrographs showing M18L (upper panels) and M18S (bottom panels) immunostaining with variant specific antibodies at various magnifications. Hippocampus and CA3 are magnified captions from the framed areas at immediate left-side images. (b–j) Confocal images and analyses from triple co-immunolabeled sections with the antibodies indicated at the top-left corners. Colors were arbitrarily assigned to maximize overlap visualization. (e–j) Rat brain sections were co-immunolabeled with antibodies against vesicular GABA (VGAT) and glutamate (VGLUT1) transporters, along with (e–f) anti-M18L or (h–i) anti-M18S, and representative images from (e and h) cortical and (f and i) hippocampal CA3 are shown. In every 6-panel composite: left and middle panels correspond to single-labeled or merged-channel images; right panels are ImageJ-generated bitmaps resulting from colocalization analyses, in which pixels mirror the intensity of colocalization (in white) between VGAT (top) or VGLUT1 (bottom) and the corresponding M18 splice variant. (g and j) Quantitative colocalization analyses of (G) M18L or (J) M18S with each of the vesicular transporters (VGAT and VGLUT1) Bars represent mean ± standard error of n = 4 rats. *p < 0.05, **p < 0.01, and ***p < 0.001 (paired t-test). Abbreviations: DG, dentate gyrus; hil, hilus; Hipp, hippocampus sg; stratum granulosum (i.e. granule cell layer); sm/l, stratum moleculare/lacunosum; so, stratum oriens; sp, stratum pyramidale (i.e. pyralmidal cell layer); sr, stratum radiatum (in CA3 also contains stratum lucidum. Scale bars: a, 200 (CA3) or 20 (cortex) μm; b, 30 μm; c, 50 (left) or 10 (right) μm; d, 30 (left) or 10 (right) μm; e and h, 10 μm; f and i, 25 μm

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