Source Paper
Alexandre Henriques, Vincent Croixmarie, David A. Priestman, Angela Rosenbohm, Sylvie Dirrig-Grosch et al.
Human Molecular Genetics • 2015
Amyotrophic lateral sclerosis (ALS) is a fatal adult-onset disease characterized by upper and lower motor neuron degeneration, muscle wasting and paralysis. Growing evidence suggests a link between changes in lipid metabolism and ALS. Here, we used UPLC/TOF-MS to survey the lipidome in SOD1(G86R) mice, a model of ALS. 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. Glucosylceramide synthase (GCS), the enzyme responsible for GlcCer biosynthesis, was up-regulated in muscle of SOD1(G86R) mice and ALS patients, and in muscle of wild-type mice after surgically induced denervation. Conversely, inhibition of GCS in wild-type mice, following transient peripheral nerve injury, reversed the overexpression of genes in muscle involved in oxidative metabolism and delayed motor recovery. GCS inhibition in SOD1(G86R) mice also affected the expression of metabolic genes and induced a loss of muscle strength and morphological deterioration of the motor endplates. These findings suggest that GSLs may play a critical role in ALS muscle pathology and could lead to the identification of new therapeutic targets.
Objective: To induce sciatic nerve crush injury in mice and study motor recovery and nerve regeneration, with comparison between pre-symptomatic and symptomatic disease states
This is a Sciatic Nerve Crush Injury protocol using mouse as the model organism. The procedure involves 10 procedural steps, 5 equipment items, 9 materials. Extracted from a 2015 paper published in Human Molecular Genetics.
Model and subjects
mouse • FVB/N • male • 75 days (pre-symptomatic group) and ~100 days (symptomatic group)
Study window
~1.4 week study window
Core workflow
Animal housing and maintenance • Pre-operative fasting • Anesthesia induction
Primary readouts
Key equipment and reagents
Verified items
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Direct vendor links
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FVB/N male mice overexpressing SOD1(G86R) maintained in animal facility with controlled environment
Note: 12 h light/dark cycle; water and A04 rodent chow provided ad libitum
“FVB/N male mice, overexpressing SOD1(G86R), were maintained in our animal facility at 23°C with a 12 h light/dark cycle. Mice had water and regular A04 rodent chow ad libitum”
Mice fasted overnight before sacrifice
“were fasted overnight before sacrifice”
Anesthetize mice with ketamine chlorohydrate and xylazine
Note: Dosages: ketamine 100 mg/kg, xylazine 5 mg/kg
“mice were anesthetized with ketamine chlorohydrate (100 mg/kg) and xylazine (5 mg/kg)”
Expose the sciatic nerve at mid-thigh level through surgical incision
Note: Performed at mid-thigh level
“The sciatic nerve was exposed at mid-thigh level”
Crush the exposed sciatic nerve using fine forceps
Note: Alternative procedure: 3-mm section can be removed with microscissors instead of crushing
“crushed with fine forceps for 30 s, or a 3-mm section removed with microscissors”
Close the skin incision with sutures
“The skin incision was sutured, and mice were allowed to recover”
Allow mice to recover from anesthesia and surgery
Note: Contralateral hind limb serves as control
“mice were allowed to recover. The hind limb, contralateral to the lesion, served as control”
Administer AMP-DNM via daily intraperitoneal injections to inhibit GCS enzymatic activity
Note: Dose: 25 mg/kg daily in 0.9% NaCl containing 5% DMSO; 10 days is typical timeframe for motor function recovery
“mice subjected to sciatic nerve crush were treated with daily intraperitoneal injections of N-(5-adamantane-1-yl-methoxy-pentyl)-deoxynojirimycin (AMP-DNM, Cayman Chemical Company, Ann Arbor, MI) for 10 days, the typical time frame required for recovery of motor function under normal conditions”
Euthanize mice by decapitation after deep anesthesia
Note: Sodium pentobarbital dose: 120 mg/kg
“Mice were killed by decapitation after deep anesthesia with 120 mg/kg sodium pentobarbital”
Rapidly dissect lumbar spinal cord and muscle tissue; freeze in liquid nitrogen and store at -80°C
Note: For immunolabeling experiments, muscle samples are fixed with 4% paraformaldehyde and stored in PBS at 4°C instead
“Lumbar spinal cord and muscle were rapidly dissected, frozen in liquid nitrogen and stored at -80°C. For immunolabeling experiments, dissected muscle samples were fixed with 4% paraformaldehyde and stored in PBS at 4°C”
This section explains what the experiment is doing, which readouts matter, what the data artifacts usually look like, and how the analysis should flow from raw capture to reported result.
To induce sciatic nerve crush injury in mice and study motor recovery and nerve regeneration, with comparison between pre-symptomatic and symptomatic disease states
Objective
To induce sciatic nerve crush injury in mice and study motor recovery and nerve regeneration, with comparison between pre-symptomatic and symptomatic disease states
Subjects
From papermouse • FVB/N • male • 75 days (pre-symptomatic group) and ~100 days (symptomatic group)
Cohort notes
From paperFVB/N male mice overexpressing SOD1(G86R); WT male littermates served as controls
Animal housing and maintenance
Pre-operative fasting (overnight)
Anesthesia induction
Sciatic nerve exposure
Motor recovery following sciatic nerve crush injury
From paperNot explicitly described in the methods section provided
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Nerve regeneration
From paperNot explicitly described in the methods section provided
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Electromyographic abnormalities
From paperNot explicitly described in the methods section provided
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Muscle expression of AChR-α
From paperNot explicitly described in the methods section provided
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Motor recovery following sciatic nerve crush injury
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Nerve regeneration
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Electromyographic abnormalities
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Muscle expression of AChR-α
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Acquisition
Collect raw experimental outputs with enough metadata to preserve sample identity, condition, and timing.
Preprocessing / cleaning
Not explicitly described in the methods section provided
Scoring or quantification
Quantify the primary readouts for this experiment: Motor recovery following sciatic nerve crush injury; Nerve regeneration; Electromyographic abnormalities; Muscle expression of AChR-α.
Statistical comparison
Statistical method not yet structured for this page.
Reporting output
Report representative outputs alongside summary comparisons for Motor recovery following sciatic nerve crush injury, Nerve regeneration, Electromyographic abnormalities, Muscle expression of AChR-α.
Source links and direct wording from the methods section for validation and deeper review.
Citation
Alexandre Henriques et al. (2015). Amyotrophic lateral sclerosis and denervation alter sphingolipids and up-regulate glucosylceramide synthase. Human Molecular Genetics
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Cayman Chemical Company
1 item with ReplicateScience direct pages
Estimated: $400.00
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Current status surfaces were computed from experiment data updated Mar 14, 2026.
Source access
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Steps
10
Evidence Quotes
24
Protocol Items
14
Linked Products
1
Canonical Sync
Pending
What this means
The completeness score reflects how much structured protocol data is present: steps, methods evidence, listed materials, linked products, and paper provenance.
Computed from the current experiment record updated Mar 14, 2026.
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Steps
10
Evidence
24
Specific Products
1/1
Canonical Sync
Pending
What this score means
The verification score reflects evidence coverage, subject detail, paper provenance, step depth, and whether linked products resolve to specific item pages instead of generic searches.
Computed from the current experiment record updated Mar 14, 2026.
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