IL-1 Receptor Knockout Study
Objective: Assessment of social defeat responses in IL-1 receptor type-1-deficient mice to evaluate IL-1 signaling in stress-induced anxiety and microglia activation
Gather these items before starting the experiment. Check off items as you prepare.
Equipment2
Materials2
Software1
Not specified • Not specified
1 item available from ConductScience
Estimated: $1,890.00
As an Amazon Associate, we earn from qualifying purchases. Product links help support this free resource.
Protocol Steps
Repeated Social Defeat Stress
Mice were subjected to repeated social defeat stress paradigm
Note: Study examined both wild-type and IL-1 receptor type-1-deficient mice
View evidence from paper
“Repeated social defeat in mice increased c-Fos staining in brain regions associated with fear and threat appraisal and promoted anxiety-like behavior”
Assessment of Anxiety-Like Behavior
Anxiety-like behavior was measured in mice following social defeat stress
Note: Anxiety-like behavior was dependent on β-adrenergic and IL-1 receptor activation
View evidence from paper
“Repeated social defeat also significantly increased the number of CD11b+/CD45high/Ly6Chigh macrophages that trafficked to the brain”
c-Fos Staining Analysis
Brain tissue was analyzed for c-Fos staining in regions associated with fear and threat appraisal
Note: c-Fos activation was increased in IL-1 receptor type-1-deficient mice but did not promote anxiety-like behavior
View evidence from paper
“Repeated social defeat increased c-Fos activation in IL-1 receptor type-1-deficient mice, but did not promote anxiety-like behavior”
Microglia and Macrophage Phenotyping
Flow cytometry analysis of inflammatory markers on microglia and macrophages including CD14, CD86, and TLR4
Note: Markers were increased on microglia and macrophages after social defeat
View evidence from paper
“Several inflammatory markers were increased on the surface of microglia (CD14, CD86, and TLR4) and macrophages (CD14 and CD86) after social defeat”
Microglia Morphology Assessment
Analysis of microglia morphology in medial amygdala, prefrontal cortex, and hippocampus
Note: Deramified microglia were increased in these brain regions following social defeat
View evidence from paper
“Repeated social defeat also increased the presence of deramified microglia in the medial amygdala, prefrontal cortex, and hippocampus”
Microglia mRNA Analysis
mRNA expression analysis of microglia for IL-1β and glucocorticoid responsive genes (GILZ and FKBP51)
Note: Social defeat increased IL-1β and reduced glucocorticoid responsive genes
View evidence from paper
“mRNA analysis of microglia indicated that repeated social defeat increased levels of interleukin (IL)-1β and reduced levels of glucocorticoid responsive genes”
Ex Vivo Microglia Stimulation
Microglia isolated from socially defeated and control mice were cultured and stimulated with lipopolysaccharide
Note: Microglia from defeated mice produced higher levels of IL-6, TNF-α, and MCP-1
View evidence from paper
“Microglia isolated from socially defeated mice and cultured ex vivo produced markedly higher levels of IL-6, tumor necrosis factor-α, and monocyte chemoattractant protein-1 after stimulation with lipopolysaccharide”
Propranolol Treatment
β-adrenergic receptor antagonist propranolol was administered to prevent stress-dependent changes
Note: Propranolol prevented stress-dependent changes in microglia and macrophages
View evidence from paper
“The stress-dependent changes in microglia and macrophages were prevented by propranolol, a β-adrenergic receptor antagonist”
IL-1 Receptor Type-1 Deficiency Analysis
Comparison of behavioral and immunological responses between IL-1 receptor type-1-deficient mice and wild-type controls
Note: IL-1 receptor type-1 deficiency prevented anxiety-like behavior and microglia activation despite increased c-Fos
View evidence from paper
“Repeated social defeat increased c-Fos activation in IL-1 receptor type-1-deficient mice, but did not promote anxiety-like behavior or microglia activation in the absence of functional IL-1 receptor type-1”