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Inhibiting SRC activity attenuates kainic-acid induced mouse epilepsy via reducing NR2B phosphorylation and
Lu Liu1, Lu Xia1, Yuxiang Li1
1Department of Neurology, Zhongshan Hospital, Fudan University, Fenglin Road, Shanghai 200032, China.
Objective:
To explore the effect of SRC activation on spontaneously recurrent seizures and to investigate the underlying mechanisms of NR2B phosphorylation.
Methods:
C57BL/6 mice were injected intrahippocampally with kainic acid (KA, 0.4 μg/25 g) to induce status epilepticus (SE). Saracatinib(STB) was used as an SRC inhibitor. Spontaneously recurrent seizures were monitored from day 7 to day 14 after the KA injection. Nissl's stain and NeuN were used to detect neuron loss and Timm stain was used to evaluate mossy fibre sprouting 14 days after KA injection. We also investigated the effect of SRC on full-length expression of NR2B. MDL28170 was used to inhibit calpain activity. Western blotting and qPCR were performed to verify phosphorylation levels and expression of SRC and NR2B 24 h after KA injection.
Results:
The duration of status epileptics in the SRC inhibitor group decreased significantly compared to the KA group 24 h after the injection of KA (P < 0.05). The application of the SRC inhibitor significantly reduced the degree of contralateral mossy fibre sprouting (P < 0.05) and improved the degree of neuron loss (P < 0.01) compared to the epilepsy group. Full-length NR2B levels in the ipsilateral hippocampus decreased in the epilepsy group (P < 0.01) compared to the sham group, and it further decreased in the STB inhibitor group (P < 0.01). The effect of the STB inhibitor was counteracted by simultaneous inhibition of SRC activity and calpain activation, while the level of full-length NR2B increased compared to the KA+STB group(P < 0.01). Reduction of NR2B cleavage by MDL28170 significantly increased the duration of epileptic status compared to the KA group (P < 0.05).
Significance:
Our data indicated that the early application of SRC inhibitors exerted protective effects on seizure severity, loss of neurons, and sprouting of mossy fibres in KA-induced mouse epilepsy. Seizure severity attenuation due to SRC inhibition was associated with the decrease of NR2B in both the phosphorylation and full-length forms.
Insights
SRC inhibitors reduce seizure severity and neuron loss in a mouse model of epilepsy. This protective effect is linked to decreased NR2B phosphorylation and full-length forms, suggesting a novel therapeutic target.
Area of Science:
- Neuroscience
- Epilepsy Research
- Molecular Biology
Background:
- Spontaneously recurrent seizures are a hallmark of epilepsy, often linked to complex molecular mechanisms.
- SRC activation and NR2B phosphorylation are implicated in neuronal excitability and seizure pathogenesis.
Purpose of the Study:
- To investigate the therapeutic potential of SRC inhibition in mitigating seizures and associated neuropathology.
- To elucidate the role of SRC-mediated NR2B phosphorylation in the development of epilepsy.
Main Methods:
- Kainic acid (KA) induced status epilepticus in C57BL/6 mice.
- SRC inhibitor (Saracatinib) and calpain inhibitor (MDL28170) were administered.
- Seizure activity, neuron loss, mossy fiber sprouting, and NR2B expression/phosphorylation were assessed.
Main Results:
- SRC inhibition significantly reduced seizure duration, neuron loss, and mossy fiber sprouting.
- Full-length NR2B levels decreased with SRC inhibition, an effect reversed by simultaneous calpain inhibition.
- NR2B cleavage reduction by MDL28170 exacerbated seizure duration.
Conclusions:
- Early SRC inhibition demonstrates neuroprotective effects in a KA-induced epilepsy model.
- The anticonvulsant effects of SRC inhibition are associated with reduced NR2B phosphorylation and full-length expression.
- Targeting SRC and its downstream pathways, including NR2B, may offer a novel therapeutic strategy for epilepsy.
