Dual pools of actin at presynaptic terminals

Bleckert, Adam; Photowala, Huzefa; Alford, Simon

doi:10.1152/jn.00789.2011

Cited by 56 publications

(60 citation statements)

References 54 publications

(147 reference statements)

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“…These two pools are differentially susceptible to pharmacological agents that disrupt F-actin structures. 68 Together, these studies provide ultrastructural, cell biological, and biochemical evidence that indicates that actin is present near synaptic vesicle pools and at the active zone.…”

Section: F-actin Is Required For Proper Development Of the Presynaptimentioning

confidence: 86%

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The actin cytoskeleton in presynaptic assembly

Nelson

Stavoe

Colón-Ramos

2013

Cell Adhesion & Migration

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Section: F-actin Is Required For Proper Development Of the Presynaptimentioning

confidence: 86%

“…56,83 Carboxypeptidase E active zone and in synaptic vesicle pools. 68 It would be interesting to determine whether different presynaptic F-actin structures could separately direct either active zone formation or synaptic vesicle clustering.…”

Section: F-actin Is Required For Proper Development Of the Presynaptimentioning

confidence: 99%

The actin cytoskeleton in presynaptic assembly

Nelson

Stavoe

Colón-Ramos

2013

Cell Adhesion & Migration

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“…Moreover, F-actin is involved in endocytic mechanisms in nerve terminals and also binds to Syn1 (Nelson et al, 2013). However, disruption of the F-actin network using the actin assembly inhibitor latrunculin-A (LatA), which sequesters actin monomers, fails to disrupt synaptic vesicle clustering or recycling (Bleckert et al, 2012; Cesca et al, 2010). We therefore asked whether the TTX-induced increase in clustering of Dyn3 might be dependent upon an intact actin cytoskeleton.…”

Section: Resultsmentioning

confidence: 99%

Differential targeting of dynamin-1 and dynamin-3 to nerve terminals during chronic suppression of neuronal activity

Calabrese

Halpain

2015

Molecular and Cellular Neuroscience

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Neurons express three closely related dynamin genes. Dynamin 1 has long been implicated in the regulation of synaptic vesicle recycling in nerve terminals, and dynamins 2 and 3 were more recently shown also to contribute to synaptic vesicle recycling in specific and distinguishable ways. In cultured hippocampal neurons we found that chronic suppression of spontaneous network activity differentially regulated the targeting of endogenous dynamins 1 and 3 to nerve terminals, while dynamin 2 was unaffected. Specifically, when neural activity was chronically silenced for 1–2 weeks by tetrodotoxin (TTX), the clustering of dynamin 1 at nerve terminals was reduced, while the clustering of dynamin 3 significantly increased. Moreover, dynamin 3 clustering was induced within hours by the sustained blockade of AMPA receptors, suggesting that AMPA receptors may function to prevent Dyn3 accumulation within nerve terminals. Clustering of dynamin 3 was induced by an antagonist of the calcium-dependent protein phosphatase calcineurin, but was not dependent upon intact actin filaments. TTX-induced clustering of Dyn3 occurred with a markedly slower time-course than the previously described clustering of synapsin 1. Potassium-induced depolarization rapidly de-clustered dynamin 3 from nerve terminals within minutes. These results, which have implications for homeostatic synapse restructuring, indicate that the three dynamins have evolved different regulatory mechanisms for trafficking to and from nerve terminals in response to changes in neural activity.

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“…The lack of any background autofluorescence and the absence of staining on postsynaptic structures or glia allows for unambiguous identification of antibody labeling at identified presynaptic terminals (Figure 5b, i). Coupled with a presynaptic marker such as Alexa-488 conjugated phalloidin (Figure 5a, ii) and (Figure 5b, ii), which labels presynaptic actin 20 , localization of immunohistochemical labeling to presynaptic terminals can be clearly demonstrated (Figure 5a, iii). This approach has been used …”

Section: Representative Resultsmentioning

confidence: 99%

Acute Dissociation of Lamprey Reticulospinal Axons to Enable Recording from the Release Face Membrane of Individual Functional Presynaptic Terminals

Ramachandran

Alford

2014

JoVE

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Synaptic transmission is an extremely rapid process. Action potential driven influx of Ca 2+ into the presynaptic terminal, through voltagegated calcium channels (VGCCs) located in the release face membrane, is the trigger for vesicle fusion and neurotransmitter release. Crucial to the rapidity of synaptic transmission is the spatial and temporal synchrony between the arrival of the action potential, VGCCs and the neurotransmitter release machinery. The ability to directly record Ca 2+ currents from the release face membrane of individual presynaptic terminals is imperative for a precise understanding of the relationship between presynaptic Ca 2+ and neurotransmitter release. Access to the presynaptic release face membrane for electrophysiological recording is not available in most preparations and presynaptic Ca 2+ entry has been characterized using imaging techniques and macroscopic current measurements -techniques that do not have sufficient temporal resolution to visualize Ca 2+ entry. The characterization of VGCCs directly at single presynaptic terminals has not been possible in central synapses and has thus far been successfully achieved only in the calyx-type synapse of the chick ciliary ganglion and in rat calyces. We have successfully addressed this problem in the giant reticulospinal synapse of the lamprey spinal cord by developing an acutely dissociated preparation of the spinal cord that yields isolated reticulospinal axons with functional presynaptic terminals devoid of postsynaptic structures. We can fluorescently label and identify individual presynaptic terminals and target them for recording. Using this preparation, we have characterized VGCCs directly at the release face of individual presynaptic terminals using immunohistochemistry and electrophysiology approaches. Ca 2+ currents have been recorded directly at the release face membrane of individual presynaptic terminals, the first such recording to be carried out at central synapses. Video LinkThe video component of this article can be found at

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Dual pools of actin at presynaptic terminals

Abstract: Bleckert A, Photowala H, Alford S. Dual pools of actin at presynaptic terminals.

Cited by 56 publications

References 54 publications

The actin cytoskeleton in presynaptic assembly

The actin cytoskeleton in presynaptic assembly

Differential targeting of dynamin-1 and dynamin-3 to nerve terminals during chronic suppression of neuronal activity

Acute Dissociation of Lamprey Reticulospinal Axons to Enable Recording from the Release Face Membrane of Individual Functional Presynaptic Terminals

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