role of the proteasome in excitotoxicity-induced cleavage of glutamic acid decarboxylase in cultured hippocampal neurons作用的蛋白酶体excitotoxicity-induced乳沟培养的海马神经元谷氨酸脱羧酶.pdfVIP

Role of the Proteasome in Excitotoxicity-Induced Cleavage of Glutamic Acid Decarboxylase in Cultured Hippocampal Neurons ´ 1. 1*.¤a ´ ˜ 1 1 1 Marcio S. Baptista , Carlos V. Melo , Mario Armelao , Dennis Herrmann , Diogo O. Pimentel , 1 1 2 ´ 3¤b 1 Graciano Leal , Margarida V. Caldeira , Ben A. Bahr , Mario Bengtson , Ramiro D. Almeida , Carlos B. Duarte1 1 Center for Neuroscience and Cell Biology, Department of Life Sciences, University of Coimbra, Coimbra, Portugal, 2 Biotechnology Research and Training Center, University of North Carolina, Pembroke, North Carolina, United States of America, 3 Department of Cancer and Cell Biology, Genomics Institute of the Novartis Research Foundation (GNF), San Diego, California, United States of America Abstract Glutamic acid decarboxylase is responsible for synthesizing GABA, the major inhibitory neurotransmitter, and exists in two isoforms—GAD65 and GAD67. The enzyme is cleaved under excitotoxic conditions, but the mechanisms involved and the functional consequences are not fully elucidated. We found that excitotoxic stimulation of cultured hippocampal neurons with glutamate leads to a time-dependent cleavage of GAD65 and GAD67 in the N-terminal region of the proteins, and decrease the corresponding mRNAs. The cleavage of GAD67 was sensitive to the proteasome inhibitors MG132, YU102 and lactacystin, and was also abrogated by the E1 ubiquitin ligase inhibitor UBEI-41. In contrast, MG132 and UBEI-41 were the only inhibitors tested that showed an effect on GAD65 cleavage. Excitotoxic stimulation with glutama