title: Simultaneous monitoring of presynaptic transmitter release and postsynaptic receptor trafficking reveals an enhancement of presynaptic activity in metabotropic glutamate receptor-mediated long-term depression.
abstract: Although the contribution of postsynaptic mechanisms to long-term synaptic plasticity has been studied extensively, understanding the contribution of presynaptic modifications to this process lags behind, primarily because of a lack of techniques with which to directly and quantifiably measure neurotransmitter release from synaptic terminals. Here, we developed a method to measure presynaptic activity through the biotinylation of vesicular transporters in vesicles fused with presynaptic membranes during neurotransmitter release. This method allowed us for the first time to selectively quantify the spontaneous or evoked release of glutamate or GABA at their respective synapses. Using this method to investigate presynaptic changes during the expression of group I metabotropic glutamate receptor (mGluR1/5)-mediated long-term depression (LTD) in cultured rat hippocampal neurons, we discovered that this form of LTD was associated with increased presynaptic release of glutamate, despite reduced miniature EPSCs measured with whole-cell recording. Moreover, we found that specific blockade of AMPA receptor (AMPAR) endocytosis with a membrane-permeable GluR2-derived peptide not only prevented the expression of LTD but also eliminated LTD-associated increase in presynaptic release. Thus, our work not only demonstrates that mGluR1/5-mediated LTD is associated with increased endocytosis of postsynaptic AMPARs but also reveals an unexpected homeostatic/compensatory increase in presynaptic release. In addition, this study indicates that biotinylation of vesicular transporters in live cultured neurons is a valuable tool for studying presynaptic function.
authors.affiliation: Brain Research Center, University of British Columbia, Vancouver, British Columbia, Canada V6T 2B5.
authors.forename: Wei
authors.initials: W
authors.lastname: Xu
authors.affiliation: Douglas Mental Health University Institute, Montreal, Quebec, Canada H4H 1R3.
authors.forename: Yiu Chung
authors.initials: YC
authors.lastname: Tse
authors.affiliation: Brain Research Center, University of British Columbia, Vancouver, British Columbia, Canada V6T 2B5.
authors.forename: Frederick A
authors.initials: FA
authors.lastname: Dobie
authors.affiliation: Graduate College of Biomedical Sciences, Western University of Health Sciences, Pomona, California 91766.
authors.forename: Michel
authors.initials: M
authors.lastname: Baudry
authors.affiliation: Brain Research Center, University of British Columbia, Vancouver, British Columbia, Canada V6T 2B5.
authors.forename: Ann Marie
authors.initials: AM
authors.lastname: Craig
authors.affiliation: Douglas Mental Health University Institute, Montreal, Quebec, Canada H4H 1R3.|Department of Psychiatry, McGill University, Montreal, Quebec, Canada H4H 1R3.
authors.forename: Tak Pan
authors.initials: TP
authors.lastname: Wong
authors.affiliation: Brain Research Center, University of British Columbia, Vancouver, British Columbia, Canada V6T 2B5.|Translational Medicine Research Center and Center for Neuropsychiatry, China Medical University Hospital and Graduate Institute of Immunology, China Medical University, Taichung, Taiwan 40447.
authors.forename: Yu Tian
authors.initials: YT
authors.lastname: Wang
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D001640:Bicuculline / Q000031:analogs & derivatives / Q000494:pharmacology; D001703:Biophysics; D001710:Biotin / Q000031:analogs & derivatives / Q000494:pharmacology; D019921:Biotinylation; D004558:Electric Stimulation; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D058787:GABA-A Receptor Antagonists / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology; D036881:Long-Term Synaptic Depression / Q000187:drug effects / Q000502:physiology*; D008734:Methoxyhydroxyphenylglycol / Q000031:analogs & derivatives / Q000494:pharmacology; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D018377:Neurotransmitter Agents / Q000378:metabolism*; D009924:Organ Culture Techniques; D018408:Patch-Clamp Techniques; D010455:Peptides / Q000494:pharmacology; D011188:Potassium / Q000494:pharmacology; D017729:Presynaptic Terminals / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects / Q000502:physiology; D011810:Quinoxalines / Q000494:pharmacology; D051381:Rats; D018091:Receptors, AMPA / Q000737:chemistry; D018094:Receptors, Metabotropic Glutamate / Q000378:metabolism*; D011990:Receptors, Transferrin / Q000378:metabolism; D026941:Sodium Channel Blockers / Q000494:pharmacology; D013388:Succinimides / Q000494:pharmacology; D050863:Synaptotagmin I / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013779:Tetrodotoxin / Q000494:pharmacology; D050598:Vesicular Glutamate Transport Protein 1 / Q000378:metabolism; D050495:Vesicular Inhibitory Amino Acid Transport Proteins / Q000378:metabolism
chemical_list: D018691:Excitatory Amino Acid Antagonists; D058787:GABA-A Receptor Antagonists; C489787:MAP2 protein, rat; D008869:Microtubule-Associated Proteins; D018377:Neurotransmitter Agents; D010455:Peptides; D011810:Quinoxalines; D018091:Receptors, AMPA; D018094:Receptors, Metabotropic Glutamate; D011990:Receptors, Transferrin; C118623:Slc32a1 protein, rat; D026941:Sodium Channel Blockers; D013388:Succinimides; D050863:Synaptotagmin I; C494312:Syt1 protein, rat; C545564:Tfrc protein, rat; D050598:Vesicular Glutamate Transport Protein 1; D050495:Vesicular Inhibitory Amino Acid Transport Proteins; C065130:sulfo-N-hydroxysuccinimide-biotin; C017069:bicuculline methiodide; D013779:Tetrodotoxin; D008734:Methoxyhydroxyphenylglycol; C056723:FG 9041; D001710:Biotin; D013755:Tetradecanoylphorbol Acetate; C104722:glutamate receptor ionotropic, AMPA 2; D011188:Potassium; C010117:3,4-dihydroxyphenylglycol; D001640:Bicuculline
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