QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

HOME  |   SEARCH  |   ARCHIVE  |   SUBSCRIBE  |   CONTACT  |   HELP

The Journal of Neuroscience, April 13, 2005, 25(15):3833-3841; doi:10.1523/JNEUROSCI.4978-04.2005

This Article Full Text Full Text (PDF) Supplemental data Submit an eLetter Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (14) Citing Articles via Google Scholar Google Scholar Articles by Yeh, E. Articles by Zhen, M. Search for Related Content PubMed PubMed Citation Articles by Yeh, E. Articles by Zhen, M.

 Previous Article  |  Next Article 

Development/Plasticity/Repair
Identification of Genes Involved in Synaptogenesis Using a Fluorescent Active Zone Marker in Caenorhabditis elegans

Edward Yeh,¹ Taizo Kawano,¹ Robby M. Weimer,³ Jean-Louis Bessereau,³ and Mei Zhen^1,2

¹Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, M5G 1X5 Canada, ²Department of Microbiology and Medical Genetics, University of Toronto, Toronto, Ontario, M5S 1A8 Canada, and ³Biologie Cellulaire de la Synapse, Ecole Normale Superieure, 75005 Paris, France

Active zones are presynaptic regions where synaptic vesicles fuse with plasma membrane to release neurotransmitters. Active zones are highly organized structurally and are functionally conserved among different species. Synapse defective-2 (SYD-2) family proteins regulate active zone morphology in Caenorhabditis elegans and Drosophila. Here, we demonstrate by immunoelectron microscopy that at C. elegans synapses, SYD-2 localizes strictly at active zones and can be used as an active zone marker when fused to green fluorescent protein (GFP). By driving expression of SYD-2::GFP fusion protein in GABAergic neurons, we are able to visualize discrete fluorescent puncta corresponding to active zones in living C. elegans. During development, the number of GABAergic synapses made by specific motoneurons increases only slightly from larvae to adult stages. In contrast, the number of SYD-2::GFP puncta doubles, suggesting that individual synapses accommodate the increasing size of their synaptic targets mainly by incorporating more active zone materials. Furthermore, we used this marker to perform a genetic screen to identify genes involved in the development of active zones. We recovered 16 mutants with altered SYD-2::GFP expression, including alleles of five genes that have been implicated previously in synapse formation or nervous-system development. Mapping of 11 additional mutants suggests that they may represent novel genes involved in active zone formation.

View larger version (44K): [in this window] [in a new window]   Figure 6. Quantification of SNB-1::GFP and SYD-2::GFP puncta during development. Using Punc-25-DsRed to label GABAergic neurons, SYD-2::GFP (hpIs3) puncta were counted in the dorsal cord region between the commissures of motoneurons VD10 and VD12. A-C, Examples of DsRed-labeled commissures meeting the dorsal cord (arrows). Scale bar, 10 µm. D, Schematic representation of this region (courtesy of E. Jorgensen). E, The results of SNB-1::GFP (juIs1) and SYD-2::GFP (hpIs3) puncta quantification within this region (as shown in A-D) are illustrated by dark filled bars. Puncta were also quantified by counting the number of puncta within a 50 µm distance at different stages during C. elegans development. These results are shown as shaded bars. Numbers for juIs1 puncta are shown in yellow, and numbers for hpIs3 puncta are shown in blue.

 

Key words: synapse; active zone; SYD-2; immunoelectron microscopy; GFP; genetic screen

---

Received Dec 7, 2004; revised February 21, 2005; accepted March 1, 2005.

This article has been cited by other articles:

				E. Yeh, T. Kawano, S. Ng, R. Fetter, W. Hung, Y. Wang, and M. Zhen Caenorhabditis elegans Innexins Regulate Active Zone Differentiation J. Neurosci., April 22, 2009; 29(16): 5207 - 5217. [Abstract] [Full Text] [PDF]

				J. E. Tanis, J. J. Moresco, R. A. Lindquist, and M. R. Koelle Regulation of Serotonin Biosynthesis by the G Proteins G{alpha}o and G{alpha}q Controls Serotonin Signaling in Caenorhabditis elegans Genetics, January 1, 2008; 178(1): 157 - 169. [Abstract] [Full Text] [PDF]

				W. Hung, C. Hwang, M. D. Po, and M. Zhen Neuronal polarity is regulated by a direct interaction between a scaffolding protein, Neurabin, and a presynaptic SAD-1 kinase in Caenorhabditis elegans Development, January 15, 2007; 134(2): 237 - 249. [Abstract] [Full Text] [PDF]

				Y. Tanizawa, A. Kuhara, H. Inada, E. Kodama, T. Mizuno, and I. Mori Inositol monophosphatase regulates localization of synaptic components and behavior in the mature nervous system of C. elegans Genes & Dev., December 1, 2006; 20(23): 3296 - 3310. [Abstract] [Full Text] [PDF]

				R. M. Weimer, E. O. Gracheva, O. Meyrignac, K. G. Miller, J. E. Richmond, and J.-L. Bessereau UNC-13 and UNC-10/Rim Localize Synaptic Vesicles to Specific Membrane Domains J. Neurosci., August 2, 2006; 26(31): 8040 - 8047. [Abstract] [Full Text] [PDF]

				B. D. Ackley, R. J. Harrington, M. L. Hudson, L. Williams, C. J. Kenyon, A. D. Chisholm, and Y. Jin The Two Isoforms of the Caenorhabditis elegans Leukocyte-Common Antigen Related Receptor Tyrosine Phosphatase PTP-3 Function Independently in Axon Guidance and Synapse Formation J. Neurosci., August 17, 2005; 25(33): 7517 - 7528. [Abstract] [Full Text] [PDF]

Home  |   Search  |   Archive  |   Subscribe  |   Contact  |   Help