Mouse ALS Model Study
Objective: Investigate the role of innate immunity and monocyte recruitment in a mouse model of ALS to correlate inflammatory responses with neuronal loss, and evaluate treatment with anti-Ly6C monoclonal antibody on disease progression
Gather these items before starting the experiment. Check off items as you prepare.
Materials1
Not explicitly stated in provided text • Not applicable • Not mentioned in provided text • Not mentioned in provided text
As an Amazon Associate, we earn from qualifying purchases. Product links help support this free resource.
Protocol Steps
Monitor disease progression in ALS mouse model
Track disease onset and progression in the mouse ALS model, monitoring for neuronal loss and immune cell changes
Note: Progressive recruitment of inflammatory monocytes to spinal cord correlates with neuronal loss
View evidence from paper
“their progressive recruitment to the spinal cord, but not brain, correlated with neuronal loss”
Characterize splenic monocyte phenotype prior to disease onset
Analyze splenic Ly6C high monocytes to identify polarized macrophage phenotype with M1 signature and increased CCR2 expression
Note: Monocytes express M1 signature including increased chemokine receptor CCR2
View evidence from paper
“Prior to disease onset, splenic Ly6C hi monocytes expressed a polarized macrophage phenotype (M1 signature), which included increased levels of chemokine receptor CCR2”
Monitor microglia changes as disease onset approaches
Assess microglia expression of CCL2 and other chemotaxis-associated molecules in spinal cord as disease onset nears
Note: Microglia-derived CCL2 leads to recruitment of monocytes to CNS; resident microglia decrease with disease progression
View evidence from paper
“As disease onset neared, microglia expressed increased CCL2 and other chemotaxis-associated molecules, which led to the recruitment of monocytes to the CNS by spinal cord–derived microglia”
Administer anti-Ly6C monoclonal antibody treatment
Treat ALS mice with anti-Ly6C mAb to modulate inflammatory monocyte response
Note: Treatment timing relative to disease onset not specified in provided text
View evidence from paper
“Treatment with anti-Ly6C mAb modulated the Ly6C hi monocyte cytokine profile, reduced monocyte recruitment to the spinal cord, diminished neuronal loss, and extended survival”
Assess treatment outcomes
Evaluate effects of anti-Ly6C mAb treatment on monocyte cytokine profile, monocyte recruitment to spinal cord, neuronal loss, and survival
Note: Multiple outcome measures assessed including survival extension
View evidence from paper
“Treatment with anti-Ly6C mAb modulated the Ly6C hi monocyte cytokine profile, reduced monocyte recruitment to the spinal cord, diminished neuronal loss, and extended survival”
Analyze human ALS monocytes for microRNA signature
Examine CD14+ CD16- monocytes from ALS patients to identify ALS-specific microRNA inflammatory signature and compare to mouse model
Note: Human monocyte signature linked to animal model findings
View evidence from paper
“In humans with ALS, the analogous monocytes (CD14 + CD16 – ) exhibited an ALS-specific microRNA inflammatory signature similar to that observed in the ALS mouse model”