{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T22:18:46Z","timestamp":1786745926521,"version":"build-2736575974"},"reference-count":51,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2026,5,22]],"date-time":"2026-05-22T00:00:00Z","timestamp":1779408000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Science and Technology of China","award":["2021ZD0202300"],"award-info":[{"award-number":["2021ZD0202300"]}]},{"award":["2021ZD0202300"],"award-info":[{"award-number":["2021ZD0202300"]}],"id":[{"id":"https:\/\/ror.org\/027s68j25","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundations of China","doi-asserted-by":"crossref","award":["32171005"],"award-info":[{"award-number":["32171005"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"award":["32171005"],"award-info":[{"award-number":["32171005"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Pregabalin (PGB) exerts its therapeutic effects by binding to the \u03b12\u03b4 auxiliary subunits of voltage-gated calcium channels and modulates synaptic transmission in the brain. However, its influence on cerebellar parallel fiber\u2013Purkinje cell (PF\u2013PC) synaptic transmission remains unclear. In the present study, we investigated the effects of PGB on PF\u2013PC synaptic transmission using whole-cell patch-clamp recording, glutamate fluorescence imaging, immunohistochemistry, co-immunoprecipitation, Western blotting, and pharmacological approaches. Micro-application of PGB to the cerebellar molecular layer induced a concentration-dependent inhibition of PF\u2013PC excitatory postsynaptic currents (EPSCs), accompanied by an increased paired-pulse ratio. The inhibitory effect of PGB on PF\u2013PC EPSCs was abolished by extracellular blockade of N-methyl-D-aspartate receptors (NMDAR) or their GluN2A subtype, as well as by disruption of \u03b12\u03b4-1\u2013NMDAR complexes, but not by intracellular NMDAR inhibition. Glutamate sensor imaging further showed that PGB markedly reduced the fluorescence intensity of glutamate release evoked by PF stimulation. In the presence of tetrodotoxin (TTX) and a gamma-aminobutyric acid type A (GABAA) receptor antagonist, PGB reduced the frequency of miniature excitatory postsynaptic currents (mEPSCs) without affecting their amplitude. The PGB-induced reduction in mEPSC frequency was fully abolished by extracellular blockade of GluN2A-containing NMDARs or disruption of \u03b12\u03b4-1\u2013NMDAR complexes. Similarly, the inhibitory effects of PGB on PF\u2013PC EPSCs and mEPSCs were eliminated by extracellular PKA inhibition, but not by intracellular protein kinase A (PKA) inhibition. Western blot analysis showed that PGB significantly increased PKA phosphorylation in the molecular layer of the cerebellar cortex. Immunoreactivity for GluN2A and \u03b12\u03b4-1 subunits was colocalized within the molecular layer and abundantly distributed around the dendrites and somata of PCs. Co-immunoprecipitation further verified that \u03b12\u03b4-1 was co-precipitated with GluN1 in cerebellar molecular layer tissue samples. The results indicate that PGB depresses glutamate release from parallel-fiber terminals in the mouse cerebellar cortex through the presynaptic \u03b12\u03b4-1-coupled GluN2A-containing NMDAR\/PKA signaling pathway, thereby attenuating PF\u2013PC synaptic transmission.<\/jats:p>","DOI":"10.3390\/ijms27114660","type":"journal-article","created":{"date-parts":[[2026,5,22]],"date-time":"2026-05-22T09:17:09Z","timestamp":1779441429000},"page":"4660","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Pregabalin Depresses Cerebellar Parallel Fiber\u2013Purkinje Cell Synaptic Transmission by Modulating Glun2a-Containing Nmda Receptors in Mice In Vitro"],"prefix":"10.3390","volume":"27","author":[{"given":"Mei-Rui","family":"Li","sequence":"first","affiliation":[{"name":"Department of Physiology and Pathophysiology, College of Medicine, Yanbian University, Yanji 133002, China"},{"name":"Institute of Brain Science, Jilin Medical University, Jilin 132013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xu-Dong","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Physiology and Pathophysiology, College of Medicine, Yanbian University, Yanji 133002, China"},{"name":"Institute of Brain Science, Jilin Medical University, Jilin 132013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Physiology and Pathophysiology, College of Medicine, Yanbian University, Yanji 133002, China"},{"name":"Institute of Brain Science, Jilin Medical University, Jilin 132013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi-Dan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Physiology and Pathophysiology, College of Medicine, Yanbian University, Yanji 133002, China"},{"name":"Institute of Brain Science, Jilin Medical University, Jilin 132013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chun-Yan","family":"Wang","sequence":"additional","affiliation":[{"name":"Institute of Brain Science, Jilin Medical University, Jilin 132013, China"},{"name":"Department of Physiology, College of Basic Medicine, Jilin Medical University, Jilin 132013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zi-Yu","family":"Zhao","sequence":"additional","affiliation":[{"name":"Institute of Brain Science, Jilin Medical University, Jilin 132013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mo","family":"Zhou","sequence":"additional","affiliation":[{"name":"Institute of Brain Science, Jilin Medical University, Jilin 132013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-6921-5028","authenticated-orcid":false,"given":"Chun-Ping","family":"Chu","sequence":"additional","affiliation":[{"name":"Institute of Brain Science, Jilin Medical University, Jilin 132013, China"},{"name":"Department of Physiology, College of Basic Medicine, Jilin Medical University, Jilin 132013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8825-4707","authenticated-orcid":false,"given":"De-Lai","family":"Qiu","sequence":"additional","affiliation":[{"name":"Department of Physiology and Pathophysiology, College of Medicine, Yanbian University, Yanji 133002, China"},{"name":"Institute of Brain Science, Jilin Medical University, Jilin 132013, China"},{"name":"Department of Physiology, College of Basic Medicine, Jilin Medical University, Jilin 132013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"17537","DOI":"10.1073\/pnas.0409066103","article-title":"Identification of the alpha2-delta-1 subunit of voltage-dependent calcium channels as a molecular target for pain mediating the analgesic actions of pregabalin","volume":"103","author":"Field","year":"2006","journal-title":"Proc. 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