Rho-Kinase/ROCK Phosphorylates PSD-93 Downstream of NMDARs to Orchestrate Synaptic Plasticity
Abstract
:1. Introduction
2. Results
2.1. Rho-Kinase Phosphorylates PSD-93 In Vitro
2.2. Rho-Kinase Phosphorylates PSD-93 at Thr612 in Striatal Slices
2.3. NMDA Induces the Phosphorylation of PSD-93 at Thr612 in Striatal Slices
2.4. Identification of PSD-93-Binding Proteins by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) Analysis
2.5. Rho-Kinase Regulates the Interaction of PSD-93 with Partner Proteins
2.6. Chemically Induced-LTP Increases the Rho-Kinase-Mediated Phosphorylation of PSD-93 and the Colocalization of PSD-93 with PSD-95
3. Discussion
3.1. Phosphorylation of PSD-93
3.2. Roles of PSD-93 Phosphorylation in PSD Complex Formation
3.3. Roles of PSD-93 Phosphorylation in Synaptic Plasticity
4. Materials and Methods
4.1. Animals
4.2. Materials
4.3. Plasmid Construction
4.4. Striatal Neuronal Culture
4.5. In Vitro Phosphorylation Assay
4.6. Striatal Slice Culture
4.7. Immunoprecipitation Assay
4.8. Mass Spectrometry
4.9. Cell Culture
4.10. GST Pull-Down Assay
4.11. Immunoblotting
4.12. Chemical LTP Assay
4.13. AAV Virus Preparation and Transfection
4.14. Immunostaining
4.15. 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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List of Phosphorylation Dependent Positively Regulated PSD-93 Binding Partners | ||||||
---|---|---|---|---|---|---|
Agap3 | Dlg1 | Dlgap1 | Dlg4 | Dlgap4 | Grin1 | Grin2a |
Grin2b | Grik5 | Iqsec2 | Nrcam | Rtn3 |
List of Phosphorylation Dependent Negatively Regulated PSD-93 Binding Partners | ||||||
---|---|---|---|---|---|---|
ADAM22 | Begain | Cacna2d1 | Cit | Cnksr2 | Dclk1 | Dlgap2 |
Dlgap3 | Griks | Kcnj4 | Lrrc7 | LGI1 | Map4 | Prph |
Shank3 | Shisa7 | SynGAP1 |
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Hossen, E.; Funahashi, Y.; Faruk, M.O.; Ahammad, R.U.; Amano, M.; Yamada, K.; Kaibuchi, K. Rho-Kinase/ROCK Phosphorylates PSD-93 Downstream of NMDARs to Orchestrate Synaptic Plasticity. Int. J. Mol. Sci. 2023, 24, 404. https://doi.org/10.3390/ijms24010404
Hossen E, Funahashi Y, Faruk MO, Ahammad RU, Amano M, Yamada K, Kaibuchi K. Rho-Kinase/ROCK Phosphorylates PSD-93 Downstream of NMDARs to Orchestrate Synaptic Plasticity. International Journal of Molecular Sciences. 2023; 24(1):404. https://doi.org/10.3390/ijms24010404
Chicago/Turabian StyleHossen, Emran, Yasuhiro Funahashi, Md. Omar Faruk, Rijwan Uddin Ahammad, Mutsuki Amano, Kiyofumi Yamada, and Kozo Kaibuchi. 2023. "Rho-Kinase/ROCK Phosphorylates PSD-93 Downstream of NMDARs to Orchestrate Synaptic Plasticity" International Journal of Molecular Sciences 24, no. 1: 404. https://doi.org/10.3390/ijms24010404