SNAP-25 gene family members differentially support secretory vesicle fusion

Swati Arora; Ingrid Saarloos; Robbelien Kooistra; Rhea van de Bospoort; Matthijs Verhage; Ruud F Toonen

doi:10.1242/jcs.201889

SNAP-25 gene family members differentially support secretory vesicle fusion

Swati Arora, Ingrid Saarloos, Robbelien Kooistra, Rhea van de Bospoort, Matthijs Verhage, Ruud F Toonen

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

Neuronal dense-core vesicles (DCVs) transport and secrete neuropeptides necessary for development, plasticity and survival, but little is known about their fusion mechanism. We show that Snap-25-null mutant (SNAP-25 KO) neurons, previously shown to degenerate after 4 days in vitro (DIV), contain fewer DCVs and have reduced DCV fusion probability in surviving neurons at DIV14. At DIV3, before degeneration, SNAP-25 KO neurons show normal DCV fusion, but one day later fusion is significantly reduced. To test if other SNAP homologs support DCV fusion, we expressed SNAP-23, SNAP-29 or SNAP-47 in SNAP-25 KO neurons. SNAP-23 and SNAP-29 rescued viability and supported DCV fusion in SNAP-25 KO neurons, but SNAP-23 did so more efficiently. SNAP-23 also rescued synaptic vesicle (SV) fusion while SNAP-29 did not. SNAP-47 failed to rescue viability and did not support DCV or SV fusion. These data demonstrate a developmental switch, in hippocampal neurons between DIV3 and DIV4, where DCV fusion becomes SNAP-25 dependent. Furthermore, SNAP-25 homologs support DCV and SV fusion and neuronal viability to variable extents - SNAP-23 most effectively, SNAP-29 less so and SNAP-47 ineffectively.

Original language	English
Pages (from-to)	1877-1889
Number of pages	13
Journal	Journal of Cell Science
Volume	130
Issue number	11
DOIs	https://doi.org/10.1242/jcs.201889
Publication status	Published - 1 Jun 2017

Funding

We thank N. Perrimon, L. Perkins and the Transgenic RNAi Project at Harvard Medical School (National Institutes of Health/National Institutes of General Medical Sciences R01-GM084947) for providing transgenic RNAi stocks. We also thank A. McHale for the software used to analyze of FISH images. We thank S. Little for his FISH protocol and C. Engbert for help implementing the protocol. We thank R. Horvitz, R. Gupta, and current and former Martin lab members for helpful discussions and feedback. This work was funded by the National Institutes of Health (R01GM105984 to A.C.M.) and the American Cancer Society (125792-RSG-14-039-01-CSM to A.C.M.). This work was supported in part by the NIH Pre-Doctoral Training Grant T32GM007287. P.W.M. acknowledges financial support from Department of Defense (DoD) through the National Defense Science and Engineering Graduate Fellowship (NDSEG) Program. This work was supported by an Alfred P. Sloan Research Fellowship (to J.D.) and by a James S. McDonnell Foundation Complex Systems Scholar Award (to J.D.). Deposited in PMC for release after 12 months.

Funders	Funder number
National Institutes of Health	T32GM007287
U.S. Department of Defense
American Cancer Society	125792-RSG-14-039-01-CSM
National Institute of General Medical Sciences	R01GM105984, R01-GM084947
Alfred P. Sloan Foundation
James S. McDonnell Foundation
Pan-Massachusetts Challenge
Harvard Medical School
National Defense Science and Engineering Graduate

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abstract = "Neuronal dense-core vesicles (DCVs) transport and secrete neuropeptides necessary for development, plasticity and survival, but little is known about their fusion mechanism. We show that Snap-25-null mutant (SNAP-25 KO) neurons, previously shown to degenerate after 4 days in vitro (DIV), contain fewer DCVs and have reduced DCV fusion probability in surviving neurons at DIV14. At DIV3, before degeneration, SNAP-25 KO neurons show normal DCV fusion, but one day later fusion is significantly reduced. To test if other SNAP homologs support DCV fusion, we expressed SNAP-23, SNAP-29 or SNAP-47 in SNAP-25 KO neurons. SNAP-23 and SNAP-29 rescued viability and supported DCV fusion in SNAP-25 KO neurons, but SNAP-23 did so more efficiently. SNAP-23 also rescued synaptic vesicle (SV) fusion while SNAP-29 did not. SNAP-47 failed to rescue viability and did not support DCV or SV fusion. These data demonstrate a developmental switch, in hippocampal neurons between DIV3 and DIV4, where DCV fusion becomes SNAP-25 dependent. Furthermore, SNAP-25 homologs support DCV and SV fusion and neuronal viability to variable extents - SNAP-23 most effectively, SNAP-29 less so and SNAP-47 ineffectively.",

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SNAP-25 gene family members differentially support secretory vesicle fusion

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