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The Journal of Neuroscience, February 11, 2004, 24(6):1507-1520; doi:10.1523/JNEUROSCI.3819-03.2004

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Development/Plasticity/Repair
Postsynaptic Density Assembly Is Fundamentally Different from Presynaptic Active Zone Assembly

Tal Bresler,¹ Mika Shapira,¹ Tobias Boeckers,² Thomas Dresbach,³ Marie Futter,⁴ Craig C. Garner,⁵ Kobi Rosenblum,⁶ Eckart D. Gundelfinger,³ and Noam E. Ziv¹

¹Rappaport Institute and the Department of Anatomy and Cell Biology, Technion Faculty of Medicine, Haifa 31096, Israel, ²Institute of Anatomy and Cell Biology, University of Ulm, Ulm 89081, Germany, ³Department of Neurochemistry and Molecular Biology, Leibniz Institute for Neurobiology, Magdeburg D-39118, Germany, ⁴Division of Physical Biochemistry, National Institute for Medical Research, London NW7 1AA, United Kingdom, ⁵Department of Psychiatry and Behavioral Science, Nancy Pritzker Laboratory, Stanford University, Palo Alto, California 94304-5485, and ⁶Faculty of Science, Haifa University, Haifa 31905, Israel

The cellular mechanisms involved in the formation of the glutamatergic postsynaptic density (PSD) are mainly unknown. Previous studies have indicated that PSD assembly may occur in situ by a gradual recruitment of postsynaptic molecules, whereas others have suggested that the PSD may be assembled from modular transport packets assembled elsewhere. Here we used cultured hippocampal neurons and live cell imaging to examine the process by which PSD molecules from different layers of the PSD are recruited to nascent postsynaptic sites. GFP-tagged NR1, the essential subunit of the NMDA receptor, and ProSAP1/Shank2 and ProSAP2/Shank3, scaffolding molecules thought to reside at deeper layers of the PSD, were recruited to new synaptic sites in gradual manner, with no obvious involvement of discernible discrete transport particles. The recruitment kinetics of these three PSD molecules were remarkably similar, which may indicate that PSD assembly rate is governed by a common upstream rate-limiting process. In contrast, the presynaptic active zone (AZ) molecule Bassoon was observed to be recruited to new presynaptic sites by means of a small number of mobile packets, in full agreement with previous studies. These findings indicate that the assembly processes of PSDs and AZs may be fundamentally different.

Key words: postsynaptic; presynaptic; synaptogenesis; ProSAP; NMDA; Bassoon

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Received Aug 15, 2003; revised November 17, 2003; accepted December 23, 2003.

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