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Hierarchical assembly of presynaptic components in defined C. elegans synapses

Nature Neuroscience volume 9, pages 1488–1498 (2006)Cite this article

Abstract

The presynaptic regions of axons accumulate synaptic vesicles, active zone proteins and periactive zone proteins. However, the rules for orderly recruitment of presynaptic components are not well understood. We systematically examined molecular mechanisms of presynaptic development in egg-laying synapses of Caenorhabditis elegans, demonstrating that two scaffolding molecules, SYD-1 and SYD-2, have key roles in presynaptic assembly. SYD-2 (liprin-α) was previously shown to regulate the size and the shape of active zones. We now show that in syd-1 and syd-2 mutants, synaptic vesicles and numerous other presynaptic proteins fail to accumulate at presynaptic sites. SYD-1 and SYD-2 function cell-autonomously at presynaptic terminals, downstream of synaptic specificity molecule SYG-1. SYD-1 is likely to act upstream of SYD-2 to positively regulate its synaptic assembly activity. These data imply a hierarchical organization of presynaptic assembly, in which transmembrane specificity molecules initiate synaptogenesis by recruiting a few key scaffolding proteins, which in turn assemble other presynaptic components.

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Figure 1: GFP- or YFP-tagged known and putative presynaptic proteins localize to the synaptic region of HSNL near the vulva.
Figure 2: Multiple presynaptic components mislocalize in syg-1(ky652) mutants.
Figure 3: SYG-1 is sufficient to recruit presynaptic components.
Figure 4: Three presynaptic components show severe defects in localizing to the synaptic region of HSNL in syd-1 and syd-2 mutants.
Figure 5: SYD-1 and SYD-2 function cell-autonomously in HSNL and SYD-2 overexpression can bypass requirement for SYD-1.
Figure 6: Some, but not all, presynaptic components fail to be trafficked out of the HSNL cell body in unc-104 mutants.
Figure 7: Epistasis analysis of syg-1, syd-1 and syd-2.

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Acknowledgements

We thank D. Chao (Stanford University) for contributing alleles of syd-1 and syd-2; the Caenorhabditis Genetics Centre, Y. Jin (University of California, Santa Cruz), the C. elegans Gene Knockout Consortium and the Japanese NBPR for strains; Y. Kohara (National Institute of Genetics, Japan) for EST cDNAs; M. Nonet (Washington University, St. Louis) for the rab-3 cDNA construct and J. Audhya (University of California, San Diego) for the mcherry cDNA; Y. Jin for communicating unpublished results; C. Gao and F. Chen for technical support; C. Garner for critical comments on the manuscript. This work was funded by the following grants to K.S.: US National Institutes of Health 1R01NS048392, the McKnight Endowment Fund, Sloan research fellowship and Whitehall Foundation fellowship.

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Authors and Affiliations

  1. Department of Biological Sciences, Stanford University, 385 Serra Mall, Stanford, 94305, California, USA

    Maulik R Patel, Emily K Lehrman, Vivian Y Poon & Kang Shen

  2. Neurosciences Program, Stanford University, 385 Serra Mall, Stanford, 94305, California, USA

    Maulik R Patel, Vivian Y Poon & Kang Shen

  3. Zilkha Neurogenetic Institute, University of Southern California, 1975 Zonal Avenue KAM-B16, 90089-9031, Los Angeles, California

    Justin G Crump

  4. Department of Medical Genetics and Microbiology, Samuel Lunenfeld Research Institute, University of Toronto, 600 University Avenue, Ontario, M5G 1X5, Canada

    Mei Zhen

  5. Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, New York, New York, 10021

    Cornelia I Bargmann

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Supplementary information

Supplementary Fig. 1

Colocalization of different presynaptic components with RAB-3. (PDF 708 kb)

Supplementary Fig. 2

Colocalization of SYD-1 and SYD-2 with RAB-3 at ectopic sites in syg-1 mutants. (PDF 391 kb)

Supplementary Fig. 3

Egg-laying defects in syd-1 and syd-2 mutant animals. (PDF 544 kb)

Supplementary Fig. 4

SYD-1 and SYD-2 are necessary for clustering of presynaptic components induced by egl-17::syg-2. (PDF 804 kb)

Supplementary Fig. 5

Presynaptic components accumulate normally in animals that are mutant for active zone components elks-1 or unc-10. (PDF 489 kb)

Supplementary Fig. 6

Dependence of some but not all synaptic components on SAD-1 for their localization to presynaptic sites. (PDF 427 kb)

Supplementary Fig. 7

A model for the hierarchical presynaptic assembly of HSNL synapses. (PDF 342 kb)

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Patel, M., Lehrman, E., Poon, V. et al. Hierarchical assembly of presynaptic components in defined C. elegans synapses. Nat Neurosci 9, 1488–1498 (2006). https://doi.org/10.1038/nn1806

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