Figure 1.
Structure and biosynthetic pathways of the major brain gangliosides [nomenclature is that of Svennerholm [40]].
Cer, ceramide; LacCer, lactosylceramide. GM1, GD1a, GD1b and GT1b comprise up to 97% of all gangliosides in the human central nervous system (boxed area).
Figure 2.
Differential distribution of gangliosides GD1b (B), GM1 (C), GD1a (E, the line demarcates the border from which the expression significantly decreases), GT1b (F) and MAG (D) in sagittal sections of adult C57Bl/6 mouse brain.
The negative control (A) was performed by omitting the primary antibody. Scale bar: 5000 µm.
Figure 3.
The expression of gangliosides GM1 (D2, E2, F2), GD1a (A’, B’ C’, D3, E3, F3, G’) and GT1b (A’’, B’’, C’’, D4, E4, F4, G’’) in mouse telencephalon and diencephalon.
Horizontal sections of olfactory bulb (A, A’, A’’; mitral cell layer of olfactory bulb is pointed out with black arrows). Coronal sections of adult mouse brain at coordinates: interaural line 3.22 mm, bregma –0.58 mm (B, B’, B’’) and interaural line 2.10 mm, bregma –1.7 mm (C, C’, C’’). Hippocampal formation is boxed in C’, arrowhead in C’ shows zona incerta and asterix in C’’ indicates medial and lateral habenular nuclei. Higher magnifications of CA1 and CA3 fields of pyramidal cell layer of hippocampus (D1–D5 and E1–E5, respectively) and granule cell layer of dentate gyrus (F1–F5). Sagittal sections of zona incerta (ZI), subthalamic nucleus (STh) and substantia nigra (SN) (G-G’’). The negative controls were performed by omitting the primary antibody (A, B, C, D1, E1, F1, G). Amy, amygdala; CA1, CA1 field of pyramidal cell layer of hippocampus; CA3, CA3 field of pyramidal cell layer of hippocampus; CPu, caudate putamen (striatum); Ctx, cortex; df, dorsal fornix; DG, dentate gyrus; EPl, external plexiform layer of olfactory bulb; fi, fimbria of the hippocampus; Gl, glomerular layer of olfactory bulb; GlA, glomerular cell layer of accessory olfactory bulb; GP, globus pallidus; GrA, granule cell layer of accessory olfactory bulb; GrO, granule cell layer of olfactory bulb; ic, internal capsule; MiA, mitral cell layer of accessory olfactory bulb; opt, optic tract; Pa, paraventricular hypothalamic nucleus; Pir, piriform cortex; Rt, reticular nucleus (prethalamus); st, stria terminalis; vhc, ventral hippocampal commissure. Scale bars = 1000 µm in A-A’’, G-G’’; 4000 µm in B-C’’ and 50 µm in D1–F5.
Figure 4.
The expression of gangliosides GM1 (F, K; red), GD1a (A, C, L; red), GD1b (G, M; red), GT1b (B, D, N; red), MAG (H, red) and MBP (I, red) in coronal sections of corpus callosum at the level of habenular nuclei (A-I) and horizontal sections of corticospinal tract in cervical spinal cord (J-N).
Commissural fibers and corticospinal tract are labeled with BDA (C-N; green). A subset of GD1a and GT1b expressing fibers are detected in the middle of corpus callosum (A, B; arrowheads). Cell nuclei are stained with DAPI (blue). The negative control is performed by omitting the primary antibody (E, J). Scale bars = 500 µm in A, B and 50 µm in C-N.
Figure 5.
The expression of major brain gangliosides in coronal sections of mouse cerebellum (A-I), red nucleus (J-M) and raphe nuclei (N-U).
Distribution of gangliosides GM1 (B, green, asterix denotes the white matter of cerebellum), GD1a (C, green), GD1b (D, green) and GT1b (E, green) in mouse cerebellum (low magnification). Double immunohistochemistry on GD1a (H, L, P, T; red) or GT1b (I, M, Q, U; red) and TH (G, H, I, K, L, M, O, P, Q, S, T, U; green) in cerebellum (F-I), red nucleus (J-M), raphe magnus nucleus (N-Q) and raphe obscurus nucleus (R-U). The negative controls were performed by omitting the anti-ganglioside antibody (A, G, K, O, S) or both anti-ganglioside and anti-TH antibody (F, J, N, R). Cell nuclei are stained blue using DAPI. gr, granular cell layer; mol, molecular layer; P, Purkinje cell layer; RMg, raphe magnus; RN, red nucleus; ROb, raphe obscurus. Scale bars = 200 µm in A-E, J-M; 50 µm in F-I and 100 µm in N-U.
Figure 6.
The distribution of gangliosides GD1a and GT1b in autonomic nuclei of brainstem (coronal sections).
Double immunohistochemistry with anti-tyrosine hydroxylase (TH) antibody (B-D, F-H, J-L, N-P, green) and anti-GD1a (C, G, K, O; red) or anti-GT1b (D, H, L, P; red). Locus coeruleus (A-D), lateral parabrachial nucleus (E-H), caudal ventrolateral medulla (I-L) and solitary nucleus (M-P). The negative controls (A, E, I, M) were performed by omitting primary antibodies. Cell nuclei are stained blue with DAPI. CVL, caudal ventrolateral medulla; LC, locus coeruleus; LPB, lateral parabrachial nucleus; Sol, solitary nucleus. Scale bar = 100 µm.
Figure 7.
The distribution of gangliosides in the nuclei of brainstem at the level of inferior olives: GM1 (B), GD1a (C), GD1b (E) and GT1b (F).
MAG is shown for comparison and is expressed in myelinated fibers (D). The negative control was performed by omitting the primary antibody (A). CI, caudal interstitial nucleus of the medial longitudinal fasciculus; ECu, external cuneate nucleus; Gi, gigantocellular reticular nucleus; icp, inferior cerebellar peduncule; IO, inferior olive; IRt, intermediate reticular nucleus; LPGi, lateral paragigantocellular nucleus; MVe, medial vestibular nucleus; PCRt, parvicellular reticular nucleus; Pr, prepositus nucleus; py, pyramidal tract; ROb, raphe obscurus nucleus; RPa, raphe pallidus nucleus; Sol, solitary nucleus; SpVe, spinal vestibular nucleus; sp5, spinal trigeminal tract; Sp5I – spinal trigeminal nucleus. Scale bar = 2000 µm.
Figure 8.
The distribution of gangliosides GM1(B, B’,B’’), GD1a (C, C’,C’’), GD1b (D, D’,D’’) and GT1b (E, E’,E’’) in transverse sections of mouse cervical (A, B, C, D, E), thoracic (A', B', C', D', E') and lumbar (A'', B'', C'', D'', E'') spinal cord.
The negative control is performed by omitting the primary antibody (A, A’, A’’). Black arrowheads denote the propriospinal white matter tracts. White arrows point to the corticospinal tract. Black arrow points to the Rexed laminae I and II. Scale bar = 500 µm.