Abstract
Impairment of synaptic function can lead to neuropsychiatric disorders collectively referred to as synaptopathies. The SNARE protein SNAP-25 is implicated in several brain pathologies and, indeed, brain areas of psychiatric patients often display reduced SNAP-25 expression. It has been recently found that acute downregulation of SNAP-25 in brain slices impairs long-term potentiation; however, the processes through which this occurs are still poorly defined. We show that in vivo acute downregulation of SNAP-25 in CA1 hippocampal region affects spine number. Consistently, hippocampal neurons from SNAP-25 heterozygous mice show reduced densities of dendritic spines and defective PSD-95 dynamics. Finally, we show that, in brain, SNAP-25 is part of a molecular complex including PSD-95 and p140Cap, with p140Cap being capable to bind to both SNAP-25 and PSD-95. These data demonstrate an unexpected role of SNAP-25 in controlling PSD-95 clustering and open the possibility that genetic reductions of the protein levels – as occurring in schizophrenia – may contribute to the pathology through an effect on postsynaptic function and plasticity.
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Abbreviations
- ADHD:
-
attention deficit hyperactive disorder
- AMPA:
-
α-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid
- AraC:
-
cytosine arabinoside
- BonT/E:
-
Botulinum toxin type E
- BSA:
-
bovine serum albumine
- Bsn:
-
Bassoon
- CA1:
-
Cornus Ammonis 1
- CTA:
-
conditioned taste aversion
- DIV:
-
days in vitro
- DTT:
-
Dithiothreitol
- EDTA:
-
Ethylenediaminetetraacetic acid
- EGTA:
-
ethylene glycol tetraacetic acid
- FACS:
-
fluorescence activated cell sorting
- FRAP:
-
fluorescence recovery after photobleaching
- GAPDH:
-
glyceraldehyde 3-phosphate dehydrogenase
- GFP:
-
green fluorescent protein
- HEK293:
-
Human Embryonic Kidney 293
- HEPES:
-
(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- Het:
-
heterozygous
- KRH:
-
Krebs’ Ringer’s-HEPES
- LTD:
-
long-term depression
- LTP:
-
long-term potentiation
- LUMIER:
-
LUminescence-based Mammalian intERactome
- mEPSC:
-
miniature excitatory postsynaptic current
- NMDA:
-
N-Methyl-D-aspartic acid or N-Methyl-D-aspartate
- NR1:
-
NMDA receptor subunit 1
- PBS:
-
phosphate-buffered saline
- PFA:
-
paraformaldehyde
- PI:
-
propidium iodide
- PLA:
-
proximity ligation assay
- PMSF:
-
phenylmethanesulfonyl fluoride
- PSD-95:
-
postsynaptic density of 95 kDa
- RFP:
-
red fluorescent protein
- shRNA:
-
short hairpin RNA
- siRNA:
-
small interference RNA
- SNAP-25:
-
synaptosomal-associated protein 25
- SNARE:
-
Soluble NSF Attachment Protein REceptor
- SRCIN1:
-
SRC kinase signaling inhibitor 1
- SV2A:
-
synaptic vesicle 2A
- Wt:
-
wild type
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Acknowledgements
We wish to acknowledge Professor Reinhard Jahn (MPI, Goettingen) for his valuable suggestions. Dr Simona Rodighiero (Fondazione Filarete, Milano, Italy) for assistance with FRAP experiments, Dr Cristina Sobacchi (Istituto Clinico Humanitas, Rozzano, Italy) for helping with C57BL/6J-GFP colony and Martina Zenkner (MDC, Berlin, Germany) for technical assistance. We would like to thank the Monzino Foundation (Milano, Italy) for its generous gift of the LSM 510 Meta and Perkin Elmer Ultraview confocal microscopes. The research leading to these results has received funding from the European Union Seventh Framework Programme under grant agreement n° HEALTH-F2-2009-241498 (‘EUROSPIN’ project) to MM, NEZ, KR, and EW; BMBF, ERA-Net Neuron II CIPRESS to JCM; by the Italian Ministry of Health (RF-2009-1545998 to MM and RF-2009-1471694 to RB) and by PRIN 2011 and Cariplo 2011-0540 to MM, and by CNR Research Project on Aging, Regione Lombardia Project MbMM-convenzione n°18099/RCC.
Author Contributions
GF designed and performed experiments and analyzed data; RM, IC, FA, DP, PT, EE, VS, ET, and AP performed experiments and analyzed data; JM PDF and provided reagents; NEZ, KR, RB, and EW discussed the data and contribute writing the paper; MM and EM designed the research and wrote the paper.
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Fossati, G., Morini, R., Corradini, I. et al. Reduced SNAP-25 increases PSD-95 mobility and impairs spine morphogenesis. Cell Death Differ 22, 1425–1436 (2015). https://doi.org/10.1038/cdd.2014.227
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DOI: https://doi.org/10.1038/cdd.2014.227
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