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 peripheral nerve injury via sciatic nerve ablation and study motor function recovery and nerve degeneration in SOD1(G86R) transgenic mice
This is a Sciatic Nerve Ablation protocol using mouse as the model organism. The procedure involves 12 procedural steps, 3 equipment items, 9 materials. Extracted from a 2015 paper published in Human Molecular Genetics.
Model and subjects
mouse • FVB/N male mice, overexpressing SOD1(G86R) • male • 75 days (pre-symptomatic group) and ~100 days (symptomatic group)
Study window
~1.4 week study window | ~12 hours hands-on
Core workflow
Animal housing and maintenance • Pre-operative fasting • Anesthesia induction
Primary readouts
Key equipment and reagents
Verified items
0
Direct vendor links
0
Use this page as an execution guide, then fall back to the source paper whenever you need exact exclusions, dosing details, or assay-specific caveats.
Confirm first
Use the page like this
Start here. The step list is optimized for running the experiment, with direct vendor links available inline when you need to source a cited item.
FVB/N male mice overexpressing SOD1(G86R) were maintained in animal facility at 23°C with 12 hour light/dark cycle. Mice had water and regular A04 rodent chow provided ad libitum.
Note: 12 hour light/dark cycle maintained throughout
“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 were fasted overnight before sacrifice
“were fasted overnight before sacrifice”
Mice were anesthetized with ketamine chlorohydrate at 100 mg/kg and xylazine at 5 mg/kg
Note: Intraperitoneal or intramuscular injection route not specified
“mice were anesthetized with ketamine chlorohydrate (100 mg/kg) and xylazine (5 mg/kg)”
The sciatic nerve was exposed at mid-thigh level through surgical incision
Note: Surgical site location: mid-thigh level
“The sciatic nerve was exposed at mid-thigh level”
The sciatic nerve was crushed with fine forceps for 30 seconds
Note: This is one of two injury methods used; crush injury was used for mice receiving AMP-DNM treatment
“crushed with fine forceps for 30 s”
A 3-mm section of the sciatic nerve was removed with microscissors
Note: This is the primary ablation method described in the experiment title
“a 3-mm section removed with microscissors”
The skin incision was sutured closed
“The skin incision was sutured, and mice were allowed to recover”
Mice were allowed to recover from anesthesia following surgery
Note: Recovery period duration not specified
“mice were allowed to recover”
Mice subjected to sciatic nerve crush were treated with daily intraperitoneal injections of AMP-DNM at 25 mg/kg in 0.9% NaCl containing 5% DMSO for 10 days
Note: Treatment duration corresponds to typical time frame required for motor function recovery under normal conditions
“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”
Mice were killed by decapitation after deep anesthesia with 120 mg/kg sodium pentobarbital
“Mice were killed by decapitation after deep anesthesia with 120 mg/kg sodium pentobarbital”
Lumbar spinal cord and muscle were rapidly dissected, frozen in liquid nitrogen and stored at −80°C
Note: Rapid freezing in liquid nitrogen performed immediately after dissection
“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
Note: Alternative to frozen storage for immunolabeling studies
“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 peripheral nerve injury via sciatic nerve ablation and study motor function recovery and nerve degeneration in SOD1(G86R) transgenic mice
Objective
To induce peripheral nerve injury via sciatic nerve ablation and study motor function recovery and nerve degeneration in SOD1(G86R) transgenic mice
Subjects
From papermouse • FVB/N male mice, overexpressing SOD1(G86R) • male • 75 days (pre-symptomatic group) and ~100 days (symptomatic group)
Cohort notes
From paperWT male littermates served as controls.
Animal housing and maintenance
Pre-operative fasting (overnight)
Anesthesia induction
Sciatic nerve exposure
Motor function recovery
From paperNot explicitly described in the provided methods section
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Nerve degeneration
From paperNot explicitly described in the provided methods section
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 provided methods section
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 provided methods section
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Motor function recovery
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 degeneration
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 provided methods section
Scoring or quantification
Quantify the primary readouts for this experiment: Motor function recovery; Nerve degeneration; 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 function recovery, Nerve degeneration, 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
“”
“”
“”
“”
Direct vendor pages are linked from the protocol above. This section stays focused on the full comparison view and the prep checklist.
Gather these items before starting the experiment. Check off items as you prepare.
Cayman Chemical Company
1 item with ReplicateScience direct pages
Estimated: $400.00
Use this section as the page quality checkpoint. It keeps section navigation, evidence access, readiness, and verification meaning in one place.
Current status surfaces were computed from experiment data updated Mar 14, 2026.
Source access
Jump back into the original paper or the methods evidence section when you need exact wording, exclusions, or method-specific caveats.
This protocol has structured steps plus evidence quotes, and is ready for canonical sync.
Steps
12
Evidence Quotes
24
Protocol Items
12
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.
Canonical Sync shows whether a ConductGraph-backed protocol is available for this experiment route right now. It is a sync-status signal, not a claim that every downstream vendor link or step detail is perfect.
Steps
12
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.
A page can have structured steps and still need review when evidence is thin, product links are generic, or canonical protocol coverage is still pending.