Coupling of synaptic vesicle fusion and retrieval constitutes a core mechanism ensuring maintenance presynaptic function. Recent studies using fast-freeze electron microscopy capacitance measurements reported an ultrafast mode endocytosis operating at physiological temperatures. Here, rat hippocampal neurons, we optically monitored single with high time resolution the vesicular glutamate transporter, synaptophysin V0a1 subunit vacuolar ATPase as probes. In this setting, could distinguish...

Synapses maintain both action potential-evoked and spontaneous neurotransmitter release; however, organization of these two forms release within an individual synapse remains unclear. Here, we used photobleaching properties iGluSnFR, a fluorescent probe that detects glutamate, to investigate the subsynaptic evoked in primary hippocampal cultures. In nonneuronal cells neuronal dendrites, iGluSnFR fluorescence is intensely photobleached recovers via diffusion nonphotobleached probes with time...

Recent studies have demonstrated that protein translation can be regulated by spontaneous excitatory neurotransmission. However, the impact of neurotransmitter release on gene transcription remains unclear. Here, we study effects balance between inhibitory and neurotransmission brain-derived neurotrophic factor (BDNF) regulation synaptic plasticity. Blockade events leads to an increase in Bdnf Npas4 through altered calcium signaling, which blocked antagonism NMDA receptors (NMDARs) or L-type...

Recent studies suggest that spontaneous and action potential-evoked neurotransmitter release processes are independently regulated. However, the mechanisms uncouple two forms of neurotransmission remain unclear. In cultured mouse rat neurons, we show C2 domain-containing protein copine-6 is localized to presynaptic terminals binds synaptobrevin2 as well other SNARE proteins in a Ca²⁺-dependent manner. interaction with selectively suppresses reaction requires tandem tryptophan...

Action potential-evoked vesicle fusion comprises the majority of neurotransmission within chemical synapses, but action potential-independent spontaneous also contributes to collection signals sent postsynaptic cell. Previous work has implicated in homeostatic synaptic scaling, few studies have selectively manipulated without substantial changes evoked study this function detail. Here we used a quadruple knockdown strategy reduce levels proteins soluble calcium-binding double C2 domain...

We showed previously that NKA (Na(+)/K(+)-ATPase) interacts with acetylated tubulin resulting in inhibition of its catalytic activity. In the present work we determined membrane-acetylated tubulin, presence detergent, behaves as an entity discrete molecular mass (320-400 kDa) during exclusion chromatography. also found microtubules assembled vitro are able to bind when incubated a detergent-solubilized membrane preparation, and isolated native have associated NKA. Furthermore, CD5...

Spontaneous neurotransmitter release is a fundamental property of synapses in which filled vesicles their content independent presynaptic action potentials (APs). Despite seemingly random nature, these spontaneous fusion events can be regulated by Ca²⁺ signaling pathways. Here, we probed the mechanisms that maintain sensitivity formed between hippocampal neurons cultured from rats both sexes. In this setting, examined potential role vesicle-associated membrane protein 4 (VAMP4),...

<ns4:p>Synaptic vesicle recycling is essential for sustained and reliable neurotransmission. A key component of synaptic the biogenesis process that observed in synapses maintains molecular identity vesicles. However, mechanisms by which vesicles are retrieved reconstituted after fusion remain unclear. The complex composition renders their rapid a daunting task. Therefore, this context, kiss-and-run type transient with plasma membrane without loss remains viable hypothesis can account...

Abstract Dynamins are GTPases required for pinching vesicles off the plasma membrane once a critical curvature is reached during endocytosis. Here, we probed dynamin function in central synapses by depleting all three isoforms postnatal hippocampal neurons down to negligible levels. We found decrease propensity of evoked neurotransmission as well reduction synaptic vesicle numbers. Recycling spontaneous or low levels activity were largely impervious depletion, while retrieval components at...

The nature of the mechanism coupling fusion and endocytosis synaptic vesicles has been long debated. Here, Chanaday Kavalali show that SNARE synaptobrevin-2 couples fast synchronous, single vesicle release to a Ca 2+ -dependent mode retrieval.

Experimental autoimmune encephalomyelitis (EAE) is an inflammatory demyelinating disease that mimics many of the clinical and pathological features multiple sclerosis. We have previously described a significant diminution in GABAergic regulation glutamate release from synaptosomes EAE rats isolated during acute stage disease. In order to explore possible metabolic pathways responsible for this alteration, work we evaluate direct effect different agonists on concomitant synapsin I...

Abstract Synapses maintain both action potential-evoked and spontaneous neurotransmitter release, however, organization of these two forms release within an individual synapse remains unclear. Here, we used photobleaching properties iGluSnFR, a fluorescent probe that detects glutamate, to investigate the subsynaptic evoked release. In non-neuronal cells neuronal dendrites, iGluSnFR fluorescence is intensely photobleached recovers via diffusion non-photobleached probes 10-seconds. After...