TRPA1-Mediated Src Family Kinases Activity Facilitates Cortical Spreading Depression Susceptibility and Trigeminovascular System Sensitization
Abstract
:1. Introduction
2. Results
2.1. Co-Expression of TRPA1 and SFKs in Cerebral Cortices of Mice
2.2. pYEEI Restored the Anti-TRPA1 Antibody-Reduced Cortical Susceptibility to CSD in Mouse Brain Slices
2.3. The Anti-TRPA1 Antibody Reduced the Level of Phosphorylated SFKs at Y416 Induced by CSD in Cytosols of Rat Cerebral Cortices
2.4. Co-Localization of TRPA1 and SFKs in Mouse TG
2.5. Saracatinib Reversed CGRP Release and IL-1β mRNA Promoted by Umbellulone in Mouse TG
2.6. Saracatinib Reduced CGRP Release Promoted by Dibutyryl-cAMP (dbcAMP), Which Is Ineffective on IL-1β mRNA Level, in Mouse TG
2.7. PKI (14–22) Amide Reduced the Level of Phosphorylated SFKs at Y416 Promoted by Umbellulone in Mouse TG
2.8. TRPA1 Did Not Form a Complex with SFKs in Rat Cerebral Cortices after CSD
3. Discussion
4. Materials and Methods
4.1. Animals
4.2. Immunohistochemistry
4.3. CSD Induction and Recording
4.3.1. Induction and Recording of CSD by Intrinsic Optical Imaging in Mouse Brain Slices and Experimental Design
4.3.2. Induction and Recording of CSD by Electrophysiology in Rats and Experimental Design
4.4. Protein Expression Analysis
4.4.1. Protein Extraction
4.4.2. Western Blot
4.5. Mouse TG Tissue Culture and Experimental Design
4.6. ELISA
4.7. qPCR
4.8. Co-Immunoprecipitation (Co-IP)
4.9. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Nie, L.; Jiang, L.; Quinn, J.P.; Grubb, B.D.; Wang, M. TRPA1-Mediated Src Family Kinases Activity Facilitates Cortical Spreading Depression Susceptibility and Trigeminovascular System Sensitization. Int. J. Mol. Sci. 2021, 22, 12273. https://doi.org/10.3390/ijms222212273
Nie L, Jiang L, Quinn JP, Grubb BD, Wang M. TRPA1-Mediated Src Family Kinases Activity Facilitates Cortical Spreading Depression Susceptibility and Trigeminovascular System Sensitization. International Journal of Molecular Sciences. 2021; 22(22):12273. https://doi.org/10.3390/ijms222212273
Chicago/Turabian StyleNie, Lingdi, Liwen Jiang, John P. Quinn, Blair D. Grubb, and Minyan Wang. 2021. "TRPA1-Mediated Src Family Kinases Activity Facilitates Cortical Spreading Depression Susceptibility and Trigeminovascular System Sensitization" International Journal of Molecular Sciences 22, no. 22: 12273. https://doi.org/10.3390/ijms222212273