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Microglial NLRP3 inflammasome activation leads to perineuronal net loss in cocultured neurons
Emre Tarakcioglu1,2, Kemal Ugur Tufekci3,4, Sermin Genc5,6,7
1Izmir Biomedicine and Genome Center, Balcova, Izmir, Turkey.
Background:
Microglia play a crucial role in maintaining the health of the central nervous system (CNS) through synaptic pruning, debris clearance, and pathogen elimination; however, sustained microglial inflammation can lead to neuronal damage. The NLRP3 inflammasome, a key regulator of inflammatory responses, has been implicated in various CNS diseases. Perineuronal nets (PNNs) are specialized structures that encase neuron somas, protect neurons from oxidative stress, and stabilize synaptic connections. While inflammatory microglia can degrade PNNs, the specific role of inflammasomes in this degradation process remains unclear.
Methods And Results:
To investigate the role of inflammasomes in degradation of PNNs, neurons were derived from the murine Neuro-2a cell line using retinoic acid. Subsequently, we induced the NLRP3 inflammasome in the murine N9 microglial cell line via exposure to lipopolysaccharide and ATP. The effect of NLRP3 activation on degradation of PNNs was examined by coculturing neurons and microglia with and without NLRP3 inhibition for 24 h. PNNs were quantified using immunofluorescence staining with Wisteria floribunda agglutinin and neurocan antibodies, both of which bind to PNNs. Our findings demonstrate that coculture with NLRP3-activated microglia reduces the percentage of WFA- and neurocan-positive neurons whereas the inhibition of NLRP3 reverses this effect.
Conclusions:
Our findings highlight the significant role of the NLRP3 inflammasome in the degradation of PNNs in inflammatory states. Moreover, our research suggests that targeting the NLRP3 inflammasome could protect PNN-positive neurons from the damaging effects of inflammation. These findings might provide insight into the inflammatory mechanisms underlying loss of PNNs.
Clinical Trial Number:
Not applicable.
Insights
The NLRP3 inflammasome in microglia drives the degradation of protective neuronal structures called perineuronal nets (PNNs). Inhibiting this inflammasome preserves PNNs and protects neurons during inflammation.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are crucial for central nervous system (CNS) health but can cause neuronal damage when inflamed.
- The NLRP3 inflammasome regulates inflammation and is implicated in CNS diseases.
- Perineuronal nets (PNNs) protect neurons, but inflammatory microglia can degrade them, with inflammasome involvement unclear.
Purpose of the Study:
- To investigate the role of inflammasomes in the degradation of PNNs.
- To determine if NLRP3 inflammasome activation in microglia affects PNN integrity.
Main Methods:
- Neurons derived from murine Neuro-2a cells; microglia from murine N9 cell line.
- NLRP3 inflammasome induced in microglia using lipopolysaccharide and ATP.
- Coculture of neurons and microglia with and without NLRP3 inhibition; PNN quantification via immunofluorescence (WFA, neurocan).
Main Results:
- Coculture with NLRP3-activated microglia reduced WFA- and neurocan-positive neurons.
- Inhibition of NLRP3 reversed the degradation of PNNs.
Conclusions:
- The NLRP3 inflammasome significantly contributes to PNN degradation in inflammatory conditions.
- Targeting the NLRP3 inflammasome may protect PNN-positive neurons from inflammatory damage.
- Findings offer insights into inflammatory mechanisms underlying PNN loss.
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