The kinetics of synaptic vesicle recycling measured at single presynaptic boutons

Ryan, Timothy A.; Réuter, H.; Wendland, Beverly; Schweizer, Felix E.; Tsien, Richard W.; Smith, Stephen J.

doi:10.1016/0896-6273(93)90081-2

Cited by 492 publications

(382 citation statements)

References 24 publications

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“…An alternative approach is to combine a functional measure of vesicle pools with subsequent ultrastructural investigation. One of the most widelyused methods exploits FM-dyes, fluorescent reporters that readily bind to lipid membrane and are taken up into recycling vesicles during endocytosis to provide a functional readout of vesicle turnover [21][22][23] . One dye variant, FM1-43FX, also efficiently drives the polymerization of diaminobenzidine (DAB) when photoactivated, leading to the formation of an osmiophilic precipitate [24][25][26][27][28][29][30][31] .…”

Section: Labelling and Visualizing Functional Vesiclesmentioning

confidence: 99%

Ultrastructural readout of functional synaptic vesicle pools in hippocampal slices based on FM dye labeling and photoconversion

et al. 2014

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(2014) Ultrastructural readout of functional synaptic vesicle pools in hippocampal slices based on FM dye labeling and photoconversion. Nature Protocols, 9 (6). pp. 1337 -1347 . ISSN 1754 -2189 This version is available from Sussex Research Online: http://sro.sussex.ac.uk/60182/ This document is made available in accordance with publisher policies and may differ from the published version or from the version of record. If you wish to cite this item you are advised to consult the publisher's version. Please see the URL above for details on accessing the published version. Copyright and reuse:Sussex Research Online is a digital repository of the research output of the University.Copyright and all moral rights to the version of the paper presented here belong to the individual author(s) and/or other copyright owners. To the extent reasonable and practicable, the material made available in SRO has been checked for eligibility before being made available.Copies of full text items generally can be reproduced, displayed or performed and given to third parties in any format or medium for personal research or study, educational, or not-for-profit purposes without prior permission or charge, provided that the authors, title and full bibliographic details are credited, a hyperlink and/or URL is given for the original metadata page and the content is not changed in any way.

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Section: Labelling and Visualizing Functional Vesiclesmentioning

confidence: 99%

Ultrastructural readout of functional synaptic vesicle pools in hippocampal slices based on FM dye labeling and photoconversion

et al. 2014

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“…26 To explore whether the palingenetic multiple axons induced by MT2 receptor activation were able to form functional synapses, we first examined the uptake and release of FM4-64 dye, which is widely used for examining the function of presynaptic axon terminals. 27 We treated rat primary hippocampal neurons at 3 div with IIK7, and measured FM4-64 uptake and release at the axons with time-lapse imaging 5 days after the treatment (8 div). Neurons with multiple axons were visualized by EGFP (Figure 5a).…”

Section: Resultsmentioning

confidence: 99%

The MT2 receptor stimulates axonogenesis and enhances synaptic transmission by activating Akt signaling

Man

et al. 2014

Cell Death Differ

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The MT2 receptor is a principal type of G protein-coupled receptor that mainly mediates the effects of melatonin. Deficits of melatonin/MT2 signaling have been found in many neurological disorders, including Alzheimer's disease, the most common cause of dementia in the elderly, suggesting that preservation of the MT2 receptor may be beneficial to these neurological disorders. However, direct evidence linking the MT2 receptor to cognition-related synaptic plasticity remains to be established. Here, we report that the MT2 receptor, but not the MT1 receptor, is essential for axonogenesis both in vitro and in vivo. We find that axon formation is retarded in MT2 receptor knockout mice, MT2-shRNA electroporated brain slices or primary neurons treated with an MT2 receptor selective antagonist. Activation of the MT2 receptor promotes axonogenesis that is associated with an enhancement in excitatory synaptic transmission in central neurons. The signaling components downstream of the MT2 receptor consist of the Akt/GSK-3β/CRMP-2 cascade. The MT2 receptor C-terminal motif binds to Akt directly. Either inhibition of the MT2 receptor or disruption of MT2 receptor-Akt binding reduces axonogenesis and synaptic transmission. Our data suggest that the MT2 receptor activates Akt/GSK-3β/CRMP-2 signaling and is necessary and sufficient to mediate functional axonogenesis and synaptic formation in central neurons. Synaptic circuits are established at the sites of axon-dendritic, axon-somatic or axon-axonal contact, in which functional axonogenesis is a critical step. 1 Axonogenesis can be regulated by many intracellular signals that involve cytoskeletal rearrangements, 2 local protein degradation, 3 as well as diffusional barriers. 4 Additionally, several extracellular neurotrophic factors and hormones have also been shown to have a role in axon guidance and synaptic formation in central neurons. 5,6 To date, the role of melatonin and its receptors in axonogenesis remains unclear. Most of the biological functions of melatonin are mediated by its two receptors, MT1 and MT2 receptors, both of them belong to the G protein-coupled receptor (GPCR) subfamily and are widely expressed throughout the central nervous system (CNS). 7 Activation of the MT2 receptor in response to melatonin is critical for controlling circadian rhythms 7 and regulation of slow wave sleep. 8,9 Early studies have shown that activation of the MT2 receptor in the retina reduces the release of dopamine, while dopamine inhibits growth cone motility and neurite outgrowth during embryonic development, 10,11 suggesting the involvement of the MT2 receptor in functional axonogenesis. In mutant mice with deficient expression of the MT2 gene, the induction of long-term potentiation (LTP) of excitatory synaptic transmission is impaired, and this impairment is closely related to deficits in learning. 12 In the hippocampus, the MT2 receptor inhibits GABA A receptor-mediated current, 13 which is implicated in the synaptic transmission. In Alzheimer's disease, expression of the MT...

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“…FM1-43 is one of the styryl dyes developed as fluorescent membrane markers, and it has been used extensively to study movement and recycling of synaptic vesicles [30][31][32][33][34]. No previous studies have explored whether FM1-43 can be used to monitor secretion in response to extracellular cations.…”

Section: Discussionmentioning

confidence: 99%

“…More FM1-43 molecules would therefore insert in the lipid bilayer, and the FM1-43 fluorescent signal increases. This model has been suggested in other cell models [30][31][32][33][34], but has the limitation of not considering the possibility of competitive binding between cations (i.e. Ca# + and La$ + ) and FM1-43.…”

Section: Discussionmentioning

confidence: 99%

“…This mechanism should therefore generate a cell model in which the secretory granules are stained by FM1-43, and exocytosis could be visualized as a loss of fluorescence. This model is suggested by studies in presynaptic neuronal boutons under sustained membrane depolarization : the dye is trapped during endocytosis in recycled synaptic vesicles of nerve terminals which become available again for exocytosis within 30 s and can be released subsequently by repeating nerve stimulation [31,39]. During such an experimental protocol, exocytosis is visualized as a diminution of fluorescence.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Changes in cytoplasmic calcium determine the secretory response to extracellular cations in human parathyroid cells: a confocal microscopy study using FM1-43 dye

Mihai

Lai

Schofield

et al. 2000

Biochemical Journal

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Whether activation of the calcium receptor (CaR) modulates secretory events was investigated by real-time fluorescence and confocal microscopy using fura 2 and FM1-43 fluorescent dye. Two paradigms were used : human parathyroid cells, which are stimulated by a step from a high to a low extracellular calcium concentration ([Ca# + ] ext ), and rMTC6-23 cells, a rat medullary thyroid carcinoma cell line whose secretion is stimulated by an increase in [Ca# + ] ext . Parathyroid cells were dispersed from parathyroid adenomas removed from 18 patients with primary hyperparathyroidism. In both cell types, incubation with FM1-43 (2 µM) resulted in staining of the plasma membranes, which was rapidly increased following changes in [Ca# + ] ext known to stimulate secretion. A high [Ca# + ] ext and lanthanum (La$ + ) decreased the membrane-associated FM1-43 fluorescence. Prolonged incubation (5-30 min) in the presence of FM1-43 resulted in accumulation of the dye in the cytoplasm, its granular distribution suggesting targeting of the secretory compartment. These data suggest that FM1-43 fluorescence is determined by :

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The kinetics of synaptic vesicle recycling measured at single presynaptic boutons

Cited by 492 publications

References 24 publications

Ultrastructural readout of functional synaptic vesicle pools in hippocampal slices based on FM dye labeling and photoconversion

Ultrastructural readout of functional synaptic vesicle pools in hippocampal slices based on FM dye labeling and photoconversion

The MT2 receptor stimulates axonogenesis and enhances synaptic transmission by activating Akt signaling

Changes in cytoplasmic calcium determine the secretory response to extracellular cations in human parathyroid cells: a confocal microscopy study using FM1-43 dye

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