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Amyotrophic lateral sclerosis and denervation alter sphingolipids and up-regulate glucosylceramide synthase.

Henriques A, Croixmarie V, Priestman DA, Rosenbohm A, Dirrig-Grosch S, D'Ambra E, Huebecker M, Hussain G, Boursier-Neyret C, Echaniz-Laguna A, Ludolph AC, Platt FM, Walther B, Spedding M, Loeffler JP, Gonzalez De Aguilar JL - Hum. Mol. Genet. (2015)

Bottom Line: Significant changes in lipid expression were evident in spinal cord and skeletal muscle before overt neuropathology.In silico analysis also revealed appreciable changes in sphingolipids including ceramides and glucosylceramides (GlcCer).HPLC analysis showed increased amounts of GlcCer and downstream glycosphingolipids (GSLs) in SOD1(G86R) muscle compared with wild-type littermates.

View Article: PubMed Central - PubMed

Affiliation: Université de Strasbourg, UMR_S 1118, Strasbourg, France, INSERM, U1118, Mécanismes Centraux et Péripheriques de la Neurodégénérescence, Strasbourg, France.

No MeSH data available.


Related in: MedlinePlus

Effect of inhibition of GCS enzymatic activity on SOD1(G86R) mice. (A) Hind limb muscle grip strength in WT and SOD1(G86R) mice in the absence or presence of AMP-DNM for 10 days. *P < 0.05 versus WT, #P < 0.05 versus untreated SOD1(G86R), n = 5–6. (B) Proportion of properly innervated neuromuscular junctions after 10 days of AMP-DNM treatment, as determined by co-labeling with anti-synaptophysin antibody and rhodamine-conjugated α-bungarotoxin. A total of 100–150 neuromuscular junctions per animal were counted, n = 4–6. Examples of normal and fragmented postsynaptic apparatus are shown in C. Number of separate postsynaptic gutters (D) and gutter intersections (E) per neuromuscular junction in mice as in A. A total of 20–25 neuromuscular junctions per animal were analyzed. **P < 0.01 versus WT, ##P < 0.01 versus untreated SOD1(G86R), n = 4. (F) Relative mRNA levels of AChR-α, AChR-ε, PPARα, LPL, FAT/CD36 PDK4 and GLUT4 in muscle of mice as in A. *P < 0.05, **P < 0.01 versus WT, #P < 0.05, ##P < 0.01 versus untreated SOD1(G86R), n = 4–6.
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DDV439F8: Effect of inhibition of GCS enzymatic activity on SOD1(G86R) mice. (A) Hind limb muscle grip strength in WT and SOD1(G86R) mice in the absence or presence of AMP-DNM for 10 days. *P < 0.05 versus WT, #P < 0.05 versus untreated SOD1(G86R), n = 5–6. (B) Proportion of properly innervated neuromuscular junctions after 10 days of AMP-DNM treatment, as determined by co-labeling with anti-synaptophysin antibody and rhodamine-conjugated α-bungarotoxin. A total of 100–150 neuromuscular junctions per animal were counted, n = 4–6. Examples of normal and fragmented postsynaptic apparatus are shown in C. Number of separate postsynaptic gutters (D) and gutter intersections (E) per neuromuscular junction in mice as in A. A total of 20–25 neuromuscular junctions per animal were analyzed. **P < 0.01 versus WT, ##P < 0.01 versus untreated SOD1(G86R), n = 4. (F) Relative mRNA levels of AChR-α, AChR-ε, PPARα, LPL, FAT/CD36 PDK4 and GLUT4 in muscle of mice as in A. *P < 0.05, **P < 0.01 versus WT, #P < 0.05, ##P < 0.01 versus untreated SOD1(G86R), n = 4–6.

Mentions: Considering the detrimental effect of the inhibition of GCS on motor axis regeneration, we hypothesized that loss of enzymatic activity would predispose SOD1(G86R) mice to develop the disease phenotype earlier. To test this hypothesis, we treated SOD1(G86R) mice with AMP-DNM for 10 days, starting at the pre-symptomatic stage. We did not observe behavioral signs of disease onset, such as impaired locomotion or abnormal hind limb extension reflexes. However, we found that SOD1(G86R) mice treated with AMP-DNM showed reduced hind limb grip strength (Fig. 8A). After treatment, the number of innervated synapses in treated and untreated mice was unchanged (Fig. 8B). In contrast, a more detailed analysis of the integrity of the postsynaptic apparatus revealed important morphological modifications (Fig. 8C). Specifically, the degree of fragmentation of the motor endplates was increased in treated SOD1(G86R) mice compared with untreated littermates (Fig. 8D). In addition, the number of gutter intersections per neuromuscular junction was decreased (Fig. 8E), which is interpreted as a loss of complexity. This morphological deterioration coincided with the presence of typical denervation markers, including increased expression of AChR-α and lowered expression of AChR-ε (Fig. 8F). As in the sciatic nerve crush paradigm, AMP-DNM abolished the stimulation of the expression of PPARα, LPL and PDK4, which are typically involved in favoring oxidative metabolism in ALS (11,12). In contrast, the expression of the glucose transporter GLUT4 appeared up-regulated (Fig. 8F), suggesting that GCS is involved in pathways of energy metabolism.Figure 8.


Amyotrophic lateral sclerosis and denervation alter sphingolipids and up-regulate glucosylceramide synthase.

Henriques A, Croixmarie V, Priestman DA, Rosenbohm A, Dirrig-Grosch S, D'Ambra E, Huebecker M, Hussain G, Boursier-Neyret C, Echaniz-Laguna A, Ludolph AC, Platt FM, Walther B, Spedding M, Loeffler JP, Gonzalez De Aguilar JL - Hum. Mol. Genet. (2015)

Effect of inhibition of GCS enzymatic activity on SOD1(G86R) mice. (A) Hind limb muscle grip strength in WT and SOD1(G86R) mice in the absence or presence of AMP-DNM for 10 days. *P < 0.05 versus WT, #P < 0.05 versus untreated SOD1(G86R), n = 5–6. (B) Proportion of properly innervated neuromuscular junctions after 10 days of AMP-DNM treatment, as determined by co-labeling with anti-synaptophysin antibody and rhodamine-conjugated α-bungarotoxin. A total of 100–150 neuromuscular junctions per animal were counted, n = 4–6. Examples of normal and fragmented postsynaptic apparatus are shown in C. Number of separate postsynaptic gutters (D) and gutter intersections (E) per neuromuscular junction in mice as in A. A total of 20–25 neuromuscular junctions per animal were analyzed. **P < 0.01 versus WT, ##P < 0.01 versus untreated SOD1(G86R), n = 4. (F) Relative mRNA levels of AChR-α, AChR-ε, PPARα, LPL, FAT/CD36 PDK4 and GLUT4 in muscle of mice as in A. *P < 0.05, **P < 0.01 versus WT, #P < 0.05, ##P < 0.01 versus untreated SOD1(G86R), n = 4–6.
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DDV439F8: Effect of inhibition of GCS enzymatic activity on SOD1(G86R) mice. (A) Hind limb muscle grip strength in WT and SOD1(G86R) mice in the absence or presence of AMP-DNM for 10 days. *P < 0.05 versus WT, #P < 0.05 versus untreated SOD1(G86R), n = 5–6. (B) Proportion of properly innervated neuromuscular junctions after 10 days of AMP-DNM treatment, as determined by co-labeling with anti-synaptophysin antibody and rhodamine-conjugated α-bungarotoxin. A total of 100–150 neuromuscular junctions per animal were counted, n = 4–6. Examples of normal and fragmented postsynaptic apparatus are shown in C. Number of separate postsynaptic gutters (D) and gutter intersections (E) per neuromuscular junction in mice as in A. A total of 20–25 neuromuscular junctions per animal were analyzed. **P < 0.01 versus WT, ##P < 0.01 versus untreated SOD1(G86R), n = 4. (F) Relative mRNA levels of AChR-α, AChR-ε, PPARα, LPL, FAT/CD36 PDK4 and GLUT4 in muscle of mice as in A. *P < 0.05, **P < 0.01 versus WT, #P < 0.05, ##P < 0.01 versus untreated SOD1(G86R), n = 4–6.
Mentions: Considering the detrimental effect of the inhibition of GCS on motor axis regeneration, we hypothesized that loss of enzymatic activity would predispose SOD1(G86R) mice to develop the disease phenotype earlier. To test this hypothesis, we treated SOD1(G86R) mice with AMP-DNM for 10 days, starting at the pre-symptomatic stage. We did not observe behavioral signs of disease onset, such as impaired locomotion or abnormal hind limb extension reflexes. However, we found that SOD1(G86R) mice treated with AMP-DNM showed reduced hind limb grip strength (Fig. 8A). After treatment, the number of innervated synapses in treated and untreated mice was unchanged (Fig. 8B). In contrast, a more detailed analysis of the integrity of the postsynaptic apparatus revealed important morphological modifications (Fig. 8C). Specifically, the degree of fragmentation of the motor endplates was increased in treated SOD1(G86R) mice compared with untreated littermates (Fig. 8D). In addition, the number of gutter intersections per neuromuscular junction was decreased (Fig. 8E), which is interpreted as a loss of complexity. This morphological deterioration coincided with the presence of typical denervation markers, including increased expression of AChR-α and lowered expression of AChR-ε (Fig. 8F). As in the sciatic nerve crush paradigm, AMP-DNM abolished the stimulation of the expression of PPARα, LPL and PDK4, which are typically involved in favoring oxidative metabolism in ALS (11,12). In contrast, the expression of the glucose transporter GLUT4 appeared up-regulated (Fig. 8F), suggesting that GCS is involved in pathways of energy metabolism.Figure 8.

Bottom Line: Significant changes in lipid expression were evident in spinal cord and skeletal muscle before overt neuropathology.In silico analysis also revealed appreciable changes in sphingolipids including ceramides and glucosylceramides (GlcCer).HPLC analysis showed increased amounts of GlcCer and downstream glycosphingolipids (GSLs) in SOD1(G86R) muscle compared with wild-type littermates.

View Article: PubMed Central - PubMed

Affiliation: Université de Strasbourg, UMR_S 1118, Strasbourg, France, INSERM, U1118, Mécanismes Centraux et Péripheriques de la Neurodégénérescence, Strasbourg, France.

No MeSH data available.


Related in: MedlinePlus