Recapture after exocytosis causes differential retention of protein in granules of bovine chromaffin cells

Perrais, David; Kleppe, Ingo; Taraska, Justin W.; Almers, Wolfhard

doi:10.1113/jphysiol.2004.064410

Cited by 157 publications

(219 citation statements)

References 56 publications

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“…The differential, and even contradictory, dependence of membrane recycling on the normal function of dynamin I may serve as a point of divergence between neuroendocrine and neuronal systems. Secondly, neuroendocrine cells utilize regulation of the fusion pore to control differential transmitter release [3,8,53], forming a critical point of control between 'breed and feed' and 'fight or flight' metabolic states. While neuronal synapses do exhibit an activity-mediated shift between 'kiss and run' and 'full collapse' vesicle cycling [21,35,54], it is not clear if a parallel physiological basis would exist in synaptic systems where vesicles do not contain a proteinacious core.…”

Section: Discussionmentioning

confidence: 99%

Dynamin I plays dual roles in the activity-dependent shift in exocytic mode in mouse adrenal chromaffin cells

Fulop

Doreian

Smith

2008

Archives of Biochemistry and Biophysics

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Under low stimulation, adrenal chromaffin cells release freely-soluble catecholamines through a restricted granule fusion pore while retaining the large neuropeptide-containing proteinacious granule core. Elevated activity causes dilation of the pore and release of all granule contents. Thus, physiological differential transmitter release is achieved through regulation of fusion pore dilation. We examined the mechanism for pore dilation utilizing a combined approach of peptide transfection, electrophysiology, electrochemistry and quantitative imaging techniques. We report that disruption of dynamin I function alters both fusion modes. Under low stimulation, interference with dynamin I does not affect granule fusion but blocks its re-internalization. In full collapse mode, disruption of dynamin I limits fusion pore dilation, but does not block membrane re-internalization. These data suggest that dynamin I is involved in both modes of exocytosis by regulating contraction or dilation of the fusion pore and thus contributes to activity-dependent differential transmitter release from the adrenal medulla.

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Section: Discussionmentioning

confidence: 99%

Dynamin I plays dual roles in the activity-dependent shift in exocytic mode in mouse adrenal chromaffin cells

Fulop

Doreian

Smith

2008

Archives of Biochemistry and Biophysics

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“…Lastly, 39% of wildtype events exhibited increased fluorescence (⌬F ϽϪ10%), which were previously termed "display" events (Perrais et al, 2004) that involve limited cargo release and persistent fusion pore opening. Syt-1 4W -expressing cells exhibited a marked shift in the distribution of events by reducing "display" events (from 39 to 8%) and increasing "release" events (from 17 to 41%; Figure 3D).…”

Section: Syt-1 Membrane Interactions Regulate the Extent Of Release Fmentioning

confidence: 99%

Synaptotagmin-1 Utilizes Membrane Bending and SNARE Binding to Drive Fusion Pore Expansion

Lynch

Gerona

Kielar

et al. 2008

MBoC

120

147

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In regulated vesicle exocytosis, SNARE protein complexes drive membrane fusion to connect the vesicle lumen with the extracellular space. The triggering of fusion pore formation by Ca 2؉ is mediated by specific isoforms of synaptotagmin (Syt), which employ both SNARE complex and membrane binding. Ca 2؉ also promotes fusion pore expansion and Syts have been implicated in this process but the mechanisms involved are unclear. We determined the role of Ca 2؉ -dependent Syt-effector interactions in fusion pore expansion by expressing Syt-1 mutants selectively altered in Ca 2؉ -dependent SNARE binding or in Ca 2؉ -dependent membrane insertion in PC12 cells that lack vesicle Syts. The release of differentsized fluorescent peptide-EGFP vesicle cargo or the vesicle capture of different-sized external fluorescent probes was used to assess the extent of fusion pore dilation. We found that PC12 cells expressing partial loss-of-function Syt-1 mutants impaired in Ca 2؉ -dependent SNARE binding exhibited reduced fusion pore opening probabilities and reduced fusion pore expansion. Cells with gain-of-function Syt-1 mutants for Ca 2؉ -dependent membrane insertion exhibited normal fusion pore opening probabilities but the fusion pores dilated extensively. The results indicate that Syt-1 uses both Ca 2؉ -dependent membrane insertion and SNARE binding to drive fusion pore expansion. INTRODUCTIONNeurotransmitter and peptide hormone secretion is mediated by the Ca 2ϩ -dependent exocytosis of vesicles at the plasma membrane. Vesicle fusion proceeds through membrane intermediates of stalk formation, fusion pore opening, and fusion pore expansion (Lindau and Alvarez de Toledo, 2003). In classical modes of vesicle exocytosis, fusion pores expand to the point where the vesicle membrane flattens on the plasma membrane (full fusion), leading to complete luminal contents release (full release). However, vesicle exocytosis can utilize alternative modes in which the fusion pore either abruptly closes (kiss-and-run) or in which the fusion pore dilates but subsequently recloses (cavicapture; Henkel and Almers, 1996). These transient modes of vesicle exocytosis lead to the partial release of luminal contents depending on the size and diffusibility of the cargo (Alvarez de Toledo et al., 1993;Barg et al., 2002;Taraska et al., 2003). Ca 2ϩ levels regulate fusion pore expansion as well as fusion pore formation (Fernandez-Chacon and Alvarez de Toledo, 1995;Hartmann and Lindau, 1995;Wang et al., 2006), but the mechanisms underlying fusion pore expansion, which is the more energetically demanding step, are poorly understood (Cohen and Melikyan, 2004).A Ca 2ϩ -dependent fusion machinery mediates the triggered formation of fusion pores. Three SNARE proteins (VAMP-2 on the vesicle with SNAP25 and syntaxin-1 on the plasma membrane) form trans complexes that promote close membrane apposition and membrane fusion (Weber et al., 1998). Members of the synaptotagmin (Syt) protein family that localize to vesicles function as Ca 2ϩ sensors that couple Ca 2ϩ rises ...

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“…As a result of these interactions and transient fusion pore opening, secretory vesicles can reseal directly after exocytosis (Holroyd et al, 2002;Taraska et al, 2003), retaining part of their cargo. This may facilitate cargo reuse and regulate the extent of cargo release (Taraska et al, 2003;An and Zenisek, 2004;Perrais et al, 2004;Scalettar, 2006). Hence, secretory granules in neuroendocrine cells undergo exocytosis through at least two distinct mechanisms: full fusion and incomplete exocytosis followed by reinternalization of the partially emptied vesicle.…”

Section: Introductionmentioning

confidence: 99%

Matrix-Dependent Local Retention of Secretory Vesicle Cargo in Cortical Neurons

Wit¹,

Toonen²,

Verhage³

2009

J. Neurosci.

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Neurons secrete many diffusible signals from synaptic and other secretory vesicles. We characterized secretion of guidance cues, neuropeptides, neurotrophins, and proteases from single secretory vesicles using pHluorin-tagged cargo in cortical neurons. Stimulation triggered transient and persistent fusion events. Transient events represented full release followed by cargo diffusion or incomplete release followed by vesicle retrieval, as previously observed in neuroendocrine cells. Unexpectedly, we also observed that certain cargo, such as Semaphorin 3A (Sema3A), was delivered at the cell surface as stable deposits. Stable deposits and transient events were observed for single cargo and both were SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) and calcium dependent. The ratio between stable and transient events did not depend on cargo size, subcellular localization (synaptic vs extrasynaptic secretion), or the presence of the extracellular matrix. Instead, the ratio is cargo specific and depends on an interaction with the vesicle matrix through a basic domain in the cargo protein. Inhibition of this interaction through deletion of the basic domain in Sema3A abolished stable deposits and rendered all events transient. Strikingly, cargo favoring transient release was stably deposited after corelease with cargo favoring stable deposit. These data argue against cargo diffusion after exocytosis as a general principle. Instead, the vesicle matrix retains secreted signals, probably for focal signaling at the cell surface.

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Recapture after exocytosis causes differential retention of protein in granules of bovine chromaffin cells

Cited by 157 publications

References 56 publications

Dynamin I plays dual roles in the activity-dependent shift in exocytic mode in mouse adrenal chromaffin cells

Dynamin I plays dual roles in the activity-dependent shift in exocytic mode in mouse adrenal chromaffin cells

Synaptotagmin-1 Utilizes Membrane Bending and SNARE Binding to Drive Fusion Pore Expansion

Matrix-Dependent Local Retention of Secretory Vesicle Cargo in Cortical Neurons

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