Ultrafast endocytosis at mouse hippocampal synapses
Open accessAbstract
To sustain neurotransmission, synaptic vesicles and their associated proteins must be recycled locally at synapses. Synaptic vesicles are thought to be regenerated approximately 20 s after fusion by the assembly of clathrin scaffolds or in approximately 1 s by the reversal of fusion pores via 'kiss-and-run' endocytosis. Here we use optogenetics to stimulate cultured hippocampal neurons with a single stimulus, rapidly freeze them after fixed intervals and examine the ultrastructure using electron microscopy--'flash-and-freeze' electron microscopy. Docked vesicles fuse and collapse into the membrane within 30 ms of the stimulus. Compensatory endocytosis occurs within 50 to 100 ms at sites flanking the active zone. Invagination is blocked by inhibition of actin polymerization, and scission is blocked by inhibiting dynamin. Because intact synaptic vesicles are not recovered, this form of recycling is not compatible with kiss-and-run endocytosis; moreover, it is 200-fold faster than clathrin-mediated endocytosis. It is likely that 'ultrafast endocytosis' is specialized to restore the surface area of the membrane rapidly.
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Synaptic vesicle endocytosis at mammalian central synapses is mediated by clathrin-independent ultrafast endocytosis.
“Several recent studies suggest that synaptic vesicle endocytosis is mediated by clathrin-independent ultrafast endocytosis at mammalian central synapses and is tightly coupled to exocytosis in time and space5–9.”
From Membrane compression by synaptic vesicle exocytosis triggers ultrafast endocytosis (2023) · cites this paper as [5]
The Abstract (page not provided) reports ultrafast endocytosis at cultured mouse hippocampal synapses, supporting the proposed pathway in a mammalian central synapse. But the Discussion explicitly says it is premature to rule out clathrin’s involvement, so the paper does not establish that the pathway is clathrin-independent. (LLM, conf=0.88)
Experimental — accuracy not yet measuredThe actin ring surrounding the active zone is likely essential for ultrafast endocytosis.
“Thus, the actin ring surrounding the active zone is likely essential for ultrafast endocytosis5.”
From Membrane compression by synaptic vesicle exocytosis triggers ultrafast endocytosis (2023) · cites this paper as [5]
The Results section (‘Actin and dynamin mediate endocytosis’; page unavailable) reports that inhibiting actin polymerization prevented endocytic structures, suggesting polymerized actin is required for ultrafast endocytosis. It also locates endocytosis lateral to the active zone, but does not identify an actin ring surrounding the zone or establish that this specific structure is essential. (LLM, conf=0.86)
Experimental — accuracy not yet measuredUltrafast endocytosis initiates as fast as 20-50 ms after exocytosis and completes within 100 ms.
“Indeed, ultrafast endocytosis initiates as fast as 20–50 ms after exocytosis and completes within 100 ms5–7.”
From Membrane compression by synaptic vesicle exocytosis triggers ultrafast endocytosis (2023) · cites this paper as [5]
The Results section “Endocytosis peaks at 100 ms” (page unavailable) reports shallow pits, deep pits, and fully internalized vesicles within 50 ms after stimulation, supporting very rapid initiation and completion of some events. It does not establish initiation specifically 20 ms after exocytosis or completion of the process within 100 ms: deep pits remained at 100 ms, and the paper says a majority of added membrane had been retrieved by then. (LLM, conf=0.83)
Experimental — accuracy not yet measuredIn ultrafast endocytosis, endocytic invaginations always form at the edge of the active zone.
“In this pathway, endocytic invaginations always form at the edge of the active zone5–7.”
From Membrane compression by synaptic vesicle exocytosis triggers ultrafast endocytosis (2023) · cites this paper as [5]
The Discussion says ultrafast endocytosis takes place at the lateral edge of the active zone. The Results report that invaginations were outside the active zone and were often observed flanking it, but do not establish that they always form at the edge. (LLM, conf=0.86)
Experimental — accuracy not yet measuredFollowing exocytosis, synaptic vesicles are recovered through clathrin-mediated, ultrafast, bulk, or kiss-and-run endocytic pathways.
“Following exocytosis, synaptic vesicles are recovered through clathrin-mediated, ultrafast, bulk, or kiss-and-run endocytic pathways.”
From Molecular Physiology of the Neuronal Synapse (2026) · cites this paper as [32]
The Introduction and Discussion describe clathrin-mediated and kiss-and-run endocytosis, and the paper reports ultrafast endocytosis after vesicle fusion. But our copy does not identify bulk endocytosis as a pathway, and the Discussion says ultrafast endocytosis is unlikely to recover intact synaptic vesicles directly. The claim therefore overstates what this source supports. (LLM, conf=0.88)
Experimental — accuracy not yet measured
Ultrafast endocytosis occurs within 50-100 milliseconds following stimulation, capturing approximately four vesicles at the very edge of the active zone, and is kinetically distinct from kiss-and-run and CME.
“Ultrafast endocytosis occurs rapidly, within 50–100 milliseconds following stimulation, capturing approximately four vesicles at the very edge of the active zone [32]. This mechanism is specialized for rapidly restoring the surface area of the presynaptic membrane and is kinetically distinct from both kiss-and-run and clathrin-mediated endocytosis [33].”
From Molecular Physiology of the Neuronal Synapse (2026) · cites this paper as [32]
The Results (“Endocytosis peaks at 100 ms”) report endocytic structures adjacent to the active zone within 50 ms and say that a large endocytic vesicle has the surface area of approximately four synaptic vesicles. The Discussion distinguishes ultrafast endocytosis from kiss-and-run, while the Abstract says it is faster than clathrin-mediated endocytosis. But the source does not say it captures four intact vesicles; it says intact synaptic vesicles are not recovered. (LLM, conf=0.88)
Experimental — accuracy not yet measuredUltrafast endocytosis is tightly coupled to exocytosis in time and space.
“Several recent studies suggest that synaptic vesicle endocytosis is mediated by clathrin-independent ultrafast endocytosis at mammalian central synapses and is tightly coupled to exocytosis in time and space5–9.”
From Membrane compression by synaptic vesicle exocytosis triggers ultrafast endocytosis (2023) · cites this paper as [5]
DMSO control did not affect ultrafast endocytosis, while Latrunculin A completely blocked it.
“Consistent with our previous work5, treatment with DMSO (control) did not affect ultrafast endocytosis, while Latrunculin A treatment completely blocked it (Fig. 2c, d; more micrographs in Supplementary Fig. 4).”
From Membrane compression by synaptic vesicle exocytosis triggers ultrafast endocytosis (2023) · cites this paper as [5]
The timing and location of endocytic pit formation in simulations quantitatively recapitulates endocytic events observed by ultrastructural analysis.
“Importantly, the timing and the location of endocytic pit formation quantitatively recapitulates endocytic events observed by ultrastructural analysis5,6.”
From Membrane compression by synaptic vesicle exocytosis triggers ultrafast endocytosis (2023) · cites this paper as [5]
F-actin is essential for the initiation of ultrafast endocytosis.
“Filamentous actin (F-actin) is essential for the initiation of ultrafast endocytosis5.”
From Membrane compression by synaptic vesicle exocytosis triggers ultrafast endocytosis (2023) · cites this paper as [5]
Latrunculin A treatment completely blocks ultrafast endocytosis, with no induction of membrane pit formation.
“When neurons are treated with Latrunculin A, which sequesters monomeric actin (G-actin) and perturbs F-actin formation12,13, ultrafast endocytosis fails completely, with no induction of membrane pit formation5.”
From Membrane compression by synaptic vesicle exocytosis triggers ultrafast endocytosis (2023) · cites this paper as [5]
Ultrafast endocytosis is compensatory: the membrane area of endocytic vesicles is roughly equal to the total surface area of synaptic vesicles exocytosed.
“Interestingly, ultrafast endocytosis is compensatory - the membrane area of endocytic vesicles is roughly equal to the total surface area of synaptic vesicles exocytosed5,6”
From Membrane compression by synaptic vesicle exocytosis triggers ultrafast endocytosis (2023) · cites this paper as [5]
Kiss-and-run is a rapid mechanism involving transient opening of a fusion pore, allowing neurotransmitter release without full collapse of the SV, with retrieval often within approximately one second.
“Kiss-and-run is a rapid mechanism that involves a transient opening of a fusion pore, allowing for neurotransmitter release without full collapse of the SV into the plasma membrane [30]. The vesicle is then rapidly retrieved, often within approximately one second [31,32].”
From Molecular Physiology of the Neuronal Synapse (2026) · cites this paper as [32]
Key proteins in CME include clathrin and dynamin, which mediates scission of the budding vesicle.
“Key proteins involved include clathrin itself and dynamin, which mediates the scission of the budding vesicle from the plasma membrane [32].”
From Molecular Physiology of the Neuronal Synapse (2026) · cites this paper as [32]
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- Authors
- Watanabe, S., Rost, B. R., Camacho-Pérez, M., Davis, M. W., Söhl-Kielczynski, B., Rosenmund, C., & Jorgensen, E. M.
- Affiliations
- Howard Hughes Medical Institute (US)
- Charité – Universitätsmedizin Berlin (DE)
- Corresponding author
- Rosenmund, Christian
- Venue
- Nature
- Published
- 2013-12-04
- Volume
- 504
- Issue
- 7479
- Pages
- 242-247
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC3957339
- OpenAlex
- W2093450191
- Cited by
- 607
- References
- 54
- Citations by year
- 2026: 26 · 2025: 40 · 2024: 44 · 2023: 47 · 2022: 53 · 2021: 44 · 2020: 53 · 2019: 62 · 2018: 51 · 2017: 54 · 2016: 36 · 2015: 58 · 2014: 38 · 2013: 1
- FWCI
- 25.72
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- DZNE Pub (submittedVersion)
- Funders
- Howard Hughes Medical Institute
- National Institute of Neurological Disorders and Stroke (R37NS034307, NS034307, R01NS034307)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
- Keywords
- synaptic vesicle recycling, clathrin-mediated endocytosis, dynamin, actin polymerization, hippocampal synapses, neurotransmission, membrane internalization, active zone, optogenetic stimulation
- MeSH terms
- Actins, Action Potentials, Animals, Endocytosis, Exocytosis, Hippocampus, Membrane Fusion, Microscopy, Electron, Synaptic Transmission, Rhodopsin, Synapses, Synaptic Vesicles, Time Factors, Dynamins, Mice
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