Role of NSF in Neurotransmitter Release: A Peptide Microinjection Study at the Crayfish Neuromuscular Junction

Parnas, I.; Rashkovan, G.; O’Connor, Vincent; El-Far, O.; Betz, Heinrich; Parnas, H.

doi:10.1152/jn.01313.2005

Cited by 5 publications

(5 citation statements)

References 41 publications

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“…Key arguments are that: (i) both NSF3 and another peptide from the external surface of the D1 domain have identical inhibitory effects on both ATPase activity and synaptic transmission; and (ii) mutation of the glycine residue, which inhibits NSF function in vivo (32,33), completely abolishes the ability of NSF3 to inhibit both ATPase activity and synaptic transmission (28). Mass spectroscopy reveals that exposure to UV light produces a peptide that is identical to noncaged NSF3 (unpublished data), so that the biochemical properties of NSF3 defined in previous work (28,31) should fully apply to uncaged cNSF3. Nonetheless, to consider possible side effects of uncaging cNSF3, we performed two control experiments.…”

Section: Resultsmentioning

confidence: 75%

“…Our strategy was based on incorporating a caging group onto a key amino acid of a peptide that blocks the ␣SNAP-stimulated ATPase activity of NSF in vitro (28)(29)(30). This peptide prevents the NSF-mediated disassembly of the SNARE complex (30) and inhibits neurotransmitter release when injected into presynaptic terminals (28,31). By using this caged peptide to perturb NSF function, we found that the amount of neurotransmitter release was inhibited with a latency ranging from 1.6 to 3.2 s. Furthermore, the kinetics of neurotransmitter release was decreased even more rapidly, with a latency of 0.2 s. These very rapid actions of the uncaged inhibitory peptide lead us to conclude that the physiologically relevant locus of NSF action in the SV cycle is immediately upstream of membrane fusion and release of neurotransmitter.…”

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confidence: 99%

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Photolysis of a caged peptide reveals rapid action of N -ethylmaleimide sensitive factor before neurotransmitter release

Kuner

Gee³

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

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The time at which the N-ethylmaleimide-sensitive factor (NSF) acts during synaptic vesicle (SV) trafficking was identified by timecontrolled perturbation of NSF function with a photoactivatable inhibitory peptide. Photolysis of this caged peptide in the squid giant presynaptic terminal caused an abrupt (0.2 s) slowing of the kinetics of the postsynaptic current (PSC) and a more gradual (2-3 s) reduction in PSC amplitude. Based on the rapid rate of these inhibitory effects relative to the speed of SV recycling, we conclude that NSF functions in reactions that immediately precede neurotransmitter release. Our results indicate the locus of SNARE protein recycling in presynaptic terminals and reveal NSF as a potential target for rapid regulation of transmitter release.caged probes ͉ exocytosis ͉ synaptic transmission ͉ synaptic vesicle cycle N eurotransmitter release relies on the precisely coordinated actions of many proteins that serve to recruit synaptic vesicles (SVs) to active zones, prepare SVs for Ca 2ϩ -dependent exocytosis, and recycle used components (1-5). At the core of these trafficking reactions lies the SNARE [soluble Nethylmaleimide sensitive factor (NSF) attachment protein receptor] complex, which consists of proteins present in SVs (v-SNAREs) and the plasma membrane (t-SNAREs) (6). It is thought that trans-SNARE complexes bridging the SV and plasma membranes bring these two membranes into close apposition and mediate membrane fusion (7,8). Because SNARE complexes are highly stable, hydrolysis of ATP by the molecular chaperone NSF (9, 10) is required to disassemble used SNARE complexes and, thereby, recycle SNARE proteins in preparation for future rounds of exocytosis (11-13). Although it is generally agreed that this action of NSF is important for neurotransmitter release, it is not clear whether NSF works before or after vesicle fusion. This distinction is critical for understanding the dynamic control of synaptic transmission by NSF and elucidating the life cycle of SNARE complexes during SV trafficking.Two models have been proposed for the timing of NSF action during neurotransmitter release (Fig. 1). SNAREs could be disassembled just before fusion, meaning that NSF is active only when needed for a fusion reaction (Fig. 1 A). This model is consistent with observations that NSF is required before vesicle fusion in several experimental systems (14-20). Alternatively, NSF could dissociate SNARE complexes immediately after neurotransmitter release (Fig. 1B). Such a postfusion action of NSF could provide an attractive mechanism for sorting of v-and t-SNAREs after fusion: in this case, newly separated v-SNAREs would be carried along as recycled SVs bud from the plasma membrane, whereas t-SNAREs would remain behind in the plasma membrane. Although experimental evidence supporting this conclusion is limited (21,22), the ability to explain SNARE sorting makes a postfusion action of NSF part of most current models of SV trafficking (8,(23)(24)(25)(26).One way to distinguish between these two alternatives ...

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Section: Resultsmentioning

confidence: 75%

mentioning

confidence: 99%

Photolysis of a caged peptide reveals rapid action of N -ethylmaleimide sensitive factor before neurotransmitter release

Kuner

Gee³

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

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“…The SNAP protein NAPB (2 A1 and 2 H2) recruits NSF to membranes for vesicle fusion and exocytosis (i.e., for neurotransmitter release) (100). NSF itself also decreased here after H2, which could have antiexcitotoxic pre-and postsynaptic consequences via decreased glutamate release (77) and decreased stabilization of AMPA glutamate receptors in the postsynaptic membrane (6), respectively. SH3GL1 decreased after A1; this protein may be needed for vesicle fusion in the presynaptic terminal (86).…”

Section: Prevention Of Excitotoxicity In Cerebellum: Decreased Excitamentioning

confidence: 99%

Compensatory proteome adjustments imply tissue-specific structural and metabolic reorganization following episodic hypoxia or anoxia in the epaulette shark (Hemiscyllium ocellatum)

Dowd

Renshaw

Cech

et al. 2010

Physiological Genomics

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The epaulette shark (Hemiscyllium ocellatum) represents an ancestral vertebrate model of episodic hypoxia and anoxia tolerance at tropical temperatures. We used two-dimensional gel electrophoresis and mass spectrometry-based proteomics approaches, combined with a suite of physiological measures, to characterize this species' responses to 1) one episode of anoxia plus normoxic recovery, 2) one episode of severe hypoxia plus recovery, or 3) two episodes of severe hypoxia plus recovery. We examined these responses in the cerebellum and rectal gland, two tissues with high ATP requirements. Sharks maintained plasma ionic homeostasis following all treatments, and activities of Na(+)/K(+)-ATPase and caspase 3/7 in both tissues were unchanged. Oxygen lack and reoxygenation elicited subtle adjustments in the proteome. Hypoxia led to more extensive proteome responses than anoxia in both tissues. The cerebellum and rectal gland exhibited treatment-specific responses to oxygen limitation consistent with one or more of several strategies: 1) neurotransmitter and receptor downregulation in cerebellum to prevent excitotoxicity, 2) cytoskeletal/membrane reorganization, 3) metabolic reorganization and more efficient intracellular energy shuttling that are more consistent with sustained ATP turnover than with long-term metabolic depression, 4) detoxification of metabolic byproducts and oxidative stress in light of continued metabolic activity, particularly following hypoxia in rectal gland, and 5) activation of prosurvival signaling. We hypothesize that neuronal morphological changes facilitate prolonged protection from excitotoxicity via dendritic spine remodeling in cerebellum (i.e., synaptic structural plasticity). These results recapitulate several highly conserved themes in the anoxia and hypoxia tolerance, preconditioning, and oxidative stress literature in a single system. In addition, several of the identified pathways and proteins suggest potentially novel mechanisms for enhancing anoxia or hypoxia tolerance in vertebrates. Overall, our data show that episodic hypoxic or anoxic exposure and recovery in the epaulette shark amplifies a constitutive suite of compensatory mechanisms that further prepares them for subsequent insults.

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“…NSF is required for the disassembly and recycling of SNARE proteins during the interval between exocytosis and endocytosis, an event important for maintaining normal neurotransmitter release in presynaptic termini (Schweizer, Dresbach et al 1998; Littleton, Barnard et al 2001; Parnas, Rashkovan et al 2006). NSF is also important for synaptic plasticity, which is believed to be the cellular mechanism for the formation of memory.…”

Section: Discussionmentioning

confidence: 99%

Enhanced expression of Pctk1, Tcf12 and Ccnd1 in hippocampus of rats: Impact on cognitive function, synaptic plasticity and pathology

Bodhinathan

et al. 2012

Neurobiology of Learning and Memory

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We previously identified a set of 50 genes that were differentially transcribed in the hippocampal CA1 region of aged, learning-impaired rats compared to aged, superior learning animals during a Morris water maze paradigm. In the current study we expressed three of these genes (Pctk1, Tcf12 and Ccnd1), which had shown increased transcription in aged, learning impaired rats, in the hippocampus of young rats using viral gene transfer and tested for learning and memory deficits at age 7–14 months. Pctk1 injected animals displayed a modest deficit in acquiring latency in both the Morris Water Maze and the reverse Morris maze. In the radial arm water maze paradigm, Pctk1, Tcf12 and Ccnd1 expressing animals all showed significant deficits in spatial working memory compared to controls. Rats injected with Ccnd1 and Tcf12, but not Pctk1, also showed a significant deficit in spatial reference memory in the radial arm water maze. Electrophysiological experiments revealed no difference in LTP in Ccnd1 and Pctk1 animals. However, LTD induced by low frequency stimulation was observed in control and Ccnd1 animals, but not in Pctk1 treated animals. In addition, neither Ccnd1 nor Pctk1 expression produced any detectable neuropathology. In contrast Tcf12 expressing animals displayed significant neurodegeneration in both CA1 and dentate gyrus. Several Tcf12 animals also developed tumors that appeared to be glioblastomas, suggesting that aberrant Tcf12 expression in the hippocampus is tumorigenic. Thus, behavioral experiments suggested that overexpression of Pctk1 and Ccnd1 produce a deficit in learning and memory, but electrophysiological experiments do not point to a simple mechanism. In contrast, the learning and memory deficits in Tcf12 animals are likely due to neuropathology associated with Tcf12 gene expression.

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Role of NSF in Neurotransmitter Release: A Peptide Microinjection Study at the Crayfish Neuromuscular Junction

Cited by 5 publications

References 41 publications

Photolysis of a caged peptide reveals rapid action of N -ethylmaleimide sensitive factor before neurotransmitter release

Photolysis of a caged peptide reveals rapid action of N -ethylmaleimide sensitive factor before neurotransmitter release

Compensatory proteome adjustments imply tissue-specific structural and metabolic reorganization following episodic hypoxia or anoxia in the epaulette shark (Hemiscyllium ocellatum)

Enhanced expression of Pctk1, Tcf12 and Ccnd1 in hippocampus of rats: Impact on cognitive function, synaptic plasticity and pathology

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