Assembly of new individual excitatory synapses: time course and temporal order of synaptic molecule recruitment.
Time-lapse microscopy, retrospective immunohistochemistry, and cultured hippocampal neurons were used to determine the time frame of individual glutamatergic synapse assembly and the temporal order in which specific molecules accumulate at new synaptic junctions. New presynaptic boutons capable of activity-evoked vesicle recycling were observed to form within 30 min of initial axodendritic contact. Clusters of the presynaptic active zone protein Bassoon were present in all new boutons. Conversely, clusters of the postsynaptic molecule SAP90/PSD-95 and glutamate receptors were found on average only approximately 45 min after such boutons were first detected. AMPA- and NMDA-type glutamate receptors displayed similar clustering kinetics. These findings suggest that glutamatergic synapse assembly can occur within 1-2 hr after initial contact and that presynaptic differentiation may precede postsynaptic differentiation.
Rappaport Institute and Department of Anatomy and Cell Biology, Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel.
H V
HV
Friedman
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T
Bresler
C C
CC
Garner
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NE
Ziv
D000818:Animals; D001369:Axons / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D003712:Dendrites / Q000502:physiology; D000076122:Disks Large Homolog 4 Protein; D019706:Excitatory Postsynaptic Potentials / Q000502:physiology; D005456:Fluorescent Dyes; D018698:Glutamic Acid / Q000502:physiology; D049452:Green Fluorescent Proteins; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D007150:Immunohistochemistry; D047908:Intracellular Signaling Peptides and Proteins; D008164:Luminescent Proteins / Q000378:metabolism; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins / Q000378:metabolism; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D051381:Rats; D017207:Rats, Sprague-Dawley; D017470:Receptors, Glutamate / Q000378:metabolism; D017661:Receptors, Presynaptic / Q000502:physiology; D011999:Recruitment, Neurophysiological / Q000502:physiology*; D013569:Synapses / Q000378:metabolism / Q000502:physiology*; D013997:Time Factors
C113688:Bsn protein, rat; D000076122:Disks Large Homolog 4 Protein; C501449:Dlg4 protein, rat; C092350:FM 4-64; D005456:Fluorescent Dyes; D047908:Intracellular Signaling Peptides and Proteins; D008164:Luminescent Proteins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D017470:Receptors, Glutamate; D017661:Receptors, Presynaptic; C033652:postsynaptic density proteins; D049452:Green Fluorescent Proteins; D018698:Glutamic Acid