[en] Mutations in the proline-rich transmembrane protein 2 (PRRT2) gene cause a wide spectrum of neurological diseases, ranging from paroxysmal kinesigenic dyskinesia (PKD) to mental retardation and epilepsy. Previously, seven PKD-related PRRT2 heterozygous mutations were identified in the Taiwanese population: P91QfsX, E199X, S202HfsX, R217PfsX, R217EfsX, R240X and R308C. This study aimed to investigate the disease-causing mechanisms of these PRRT2 mutations. We first documented that Prrt2 was localized at the pre- and post-synaptic membranes with a close spatial association with SNAP25 by synaptic membrane fractionation and immunostaining of the rat neurons. Our results then revealed that the six truncating Prrt2 mutants were accumulated in the cytoplasm and thus failed to target to the cell membrane; the R308C missense mutant had significantly reduced protein expression, suggesting loss-of function effects generated by these mutations. Using in utero electroporation of shRNA into cortical neurons, we further found that knocking down Prrt2 expression in vivo resulted in a delay in neuronal migration during embryonic development and a marked decrease in synaptic density after birth. These pathologic effects and novel disease-causing mechanisms may contribute to the severe clinical symptoms in PRRT2-related diseases.
Disciplines :
Genetics & genetic processes
Author, co-author :
Liu, Yo-Tsen; Department of Neurology, Neurological Institute, Taipei Veterans General Hospital, Taipei, Taiwan ; Department of Neurology, National Yang-Ming University School of Medicine, Taipei, Taiwan
Nian, Fang-Shin ; Université de Liège - ULiège > GIGA > GIGA Neurosciences - Molecular Regulation of Neurogenesis ; Program in Molecular Medicine, National Yang-Ming University and Academia Sinica, Taipei, Taiwan ; Institute of Brain Science, National Yang-Ming University, Taipei, Taiwan
Chou, Wan-Ju; Institute of Brain Science, National Yang-Ming University, Taipei, Taiwan
Tai, Chin-Yin; Istitute of Pharmaceutics, Development Center for Biotechnology, New Taipei City, Taiwan
Kwan, Shang-Yeong; Department of Neurology, Neurological Institute, Taipei Veterans General Hospital, Taipei, Taiwan ; Department of Neurology, National Yang-Ming University School of Medicine, Taipei, Taiwan
Chen, Chien; Department of Neurology, Neurological Institute, Taipei Veterans General Hospital, Taipei, Taiwan ; Department of Neurology, National Yang-Ming University School of Medicine, Taipei, Taiwan
Kuo, Pei-Wen; Institute of Brain Science, National Yang-Ming University, Taipei, Taiwan
Lin, Po-Hsi; Department of Neurology, National Yang-Ming University School of Medicine, Taipei, Taiwan
Chen, Chin-Yi; Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan
Huang, Chia-Wei; Institute of Brain Science, National Yang-Ming University, Taipei, Taiwan
Lee, Yi-Chung; Department of Neurology, Neurological Institute, Taipei Veterans General Hospital, Taipei, Taiwan ; Department of Neurology, National Yang-Ming University School of Medicine, Taipei, Taiwan ; Brain Research Center, National Yang-Ming University, Taipei, Taiwan
Soong, Bing-Wen; Department of Neurology, Neurological Institute, Taipei Veterans General Hospital, Taipei, Taiwan ; Department of Neurology, National Yang-Ming University School of Medicine, Taipei, Taiwan ; Brain Research Center, National Yang-Ming University, Taipei, Taiwan
Tsai, Jin-Wu; Institute of Brain Science, National Yang-Ming University, Taipei, Taiwan ; Brain Research Center, National Yang-Ming University, Taipei, Taiwan ; Biophotonics and Molecular Imaging Research Center, National Yang-Ming University, Taipei, Taiwan
YTL is grateful for the funding from the National Health Research Institutes (NHRI-EX105-10507EC) and Taipei Veterans General Hospital (V104B-015 and V105C-166). She also thank for the Clinical Research Core Laboratory of Taipei Veterans General Hospital for providing experimental space and facilities. CYT is grateful to the Development Center for Biotechnology and Academia Sinica for support. BWS is grateful for the funding support from the Ministry of Science and Technology (MOST 103-2314-B-010-049-MY3, MOST 104-2745-B-010-004, MOST 104-2745-B-010-002), Brain Research Center of National Yang-Ming University (104AC-B19) and Taipei Veterans General Hospital (V103C-109, V104E9-006). JWT is grateful to the Ministry of Science and Technology (101-2320-B-010-077-MY2, 103-2911-I-010-504, 103-2628-B-010-002- MY3, 104-2633-H-010-001, and 105-2633-B-009-003), Taiwan National Health Research Institutes (NHRIEX103-10314NC), and Academia Sinica, Taiwan (AS-104-TP-B09).
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