Description
Src Family Kinases (SFKs) play a crucial role in modulating neuroinflammation and have been linked to the regulation of neuroinflammatory gene expression - a central mechanism in migraine pathology. While SFKs are recognized for their cytosolic signalling functions, little is known regarding their nuclear roles in migraine. This study explores the nuclear localization of SFKs and their function in regulating the expression of the critical chemokine C-C motif ligand 2 (CCL2), in a rat model of migraine induced by cortical spreading depression (CSD).Specific objectives are to: (i) Examine nuclear SFKs localization in cerebral cortices of rats and whether their expression is affected by CSD. (ii) Determine the relationship between SFKs activity and gene expression of CCL2, as well as the involvement of nuclear H3K4me3 enrichment, a chromatin openness marker, at the promoter of CCL2.
Adult male Sprague-Dawley rats were used, and animal procedures were approved by the University Research Ethics Review Panel and conducted in accordance with national and provincial guidelines. CSD was induced by K+-medium in the right cortices of rats and recorded by electrophysiological methods. Immunofluorescent was applied for examining SFKs localization in astrocytes and neurons; Western Blot, qPCR and Chromatin immunoprecipitation-qPCR analysis were used for detecting SFKs activity, CCL2 gene expression and nuclear H3K4me3 enrichment at CCL2 promoter in the ipsilateral cerebral cortex tissue of rats, followed by correlation analysis.
Immunofluorescent analysis revealed nuclear SFK localization, which was markedly increased following CSD induction in astrocytes, and to a lesser extent in neurons in the cerebral cortices of rats. A pronounced elevation of cytosolic SFK activity, CCL2 gene expression, and nuclear H3K4me3 enrichment at the CCL2 promoter was observed 30 minutes post-CSD induction in the ipsilateral cerebral cortices. Correlation analysis further showed a positive relationship between cytosolic SFK activity and nuclear H3K4me3 enrichment, indicating that CSD-induced enhancement of chromatin accessibility on the inflammatory gene involves SFK activity.
These data demonstrate that CSD can trigger nuclear translocation of SFKs predominantly in astrocytes, which promotes inflammatory transcription via chromatin remodeling. The findings bring a new insight into the role of SFKs, the druggable target in migraine, as a novel transcriptional regulator in the neuroinflammation-associated disease mechanism.
| Period | 10 Sept 2025 → 13 Sept 2025 |
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| Held at | International Headache Society, United Kingdom |