Spatial distribution and temporal evolution of DRONPA-fused SNAP25 clusters in adrenal chromaffin cells

Yasuko Antoku; Peter Dedecker; Paulo César da Silva Pinheiro; Tom André Jos Vosch; Jakob Balslev Sørensen

doi:10.1039/c4pp00423j

Spatial distribution and temporal evolution of DRONPA-fused SNAP25 clusters in adrenal chromaffin cells

Yasuko Antoku, Peter Dedecker, Paulo César da Silva Pinheiro, Tom André Jos Vosch, Jakob Balslev Sørensen

4 Citations (Scopus)

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Abstract

Sub-diffraction imaging of plasma membrane localized proteins, such as the SNARE (Soluble NSF Attachment Protein Receptor) proteins involved in exocytosis, in fixed cells have resulted in images with high spatial resolution, at the expense of dynamical information. Here, we have imaged localized fluorescence bursts of DRONPA-fused SNAP-25 molecules in live chromaffin cells by Total Internal Reflection Fluorescence (TIRF) imaging. We find that this method allows tracking protein cluster dynamics over relatively long times (∼20 min.), partly due to the diffusion into the TIRF field of fresh molecules, making possible the simultaneous identification of cluster size, location and temporal evolution. The results indicate that the DRONPA-fused SNAP-25 clusters display rich dynamics, going from staying constant to disappearing and reappearing in specific cluster domains within minutes.

Original language	English
Journal	Photochemical & Photobiological Sciences
Volume	14
Issue number	5
Pages (from-to)	1005-1012
Number of pages	8
ISSN	1474-905X
DOIs	https://doi.org/10.1039/c4pp00423j
Publication status	Published - 1 May 2015

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title = "Spatial distribution and temporal evolution of DRONPA-fused SNAP25 clusters in adrenal chromaffin cells",

abstract = "Sub-diffraction imaging of plasma membrane localized proteins, such as the SNARE (Soluble NSF Attachment Protein Receptor) proteins involved in exocytosis, in fixed cells have resulted in images with high spatial resolution, at the expense of dynamical information. Here, we have imaged localized fluorescence bursts of DRONPA-fused SNAP-25 molecules in live chromaffin cells by Total Internal Reflection Fluorescence (TIRF) imaging. We find that this method allows tracking protein cluster dynamics over relatively long times (∼20 min.), partly due to the diffusion into the TIRF field of fresh molecules, making possible the simultaneous identification of cluster size, location and temporal evolution. The results indicate that the DRONPA-fused SNAP-25 clusters display rich dynamics, going from staying constant to disappearing and reappearing in specific cluster domains within minutes.",

author = "Yasuko Antoku and Peter Dedecker and {da Silva Pinheiro}, {Paulo C{\'e}sar} and Vosch, {Tom Andr{\'e} Jos} and S{\o}rensen, {Jakob Balslev}",

year = "2015",

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doi = "10.1039/c4pp00423j",

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T1 - Spatial distribution and temporal evolution of DRONPA-fused SNAP25 clusters in adrenal chromaffin cells

AU - Antoku, Yasuko

AU - Dedecker, Peter

AU - da Silva Pinheiro, Paulo César

AU - Vosch, Tom André Jos

AU - Sørensen, Jakob Balslev

PY - 2015/5/1

Y1 - 2015/5/1

N2 - Sub-diffraction imaging of plasma membrane localized proteins, such as the SNARE (Soluble NSF Attachment Protein Receptor) proteins involved in exocytosis, in fixed cells have resulted in images with high spatial resolution, at the expense of dynamical information. Here, we have imaged localized fluorescence bursts of DRONPA-fused SNAP-25 molecules in live chromaffin cells by Total Internal Reflection Fluorescence (TIRF) imaging. We find that this method allows tracking protein cluster dynamics over relatively long times (∼20 min.), partly due to the diffusion into the TIRF field of fresh molecules, making possible the simultaneous identification of cluster size, location and temporal evolution. The results indicate that the DRONPA-fused SNAP-25 clusters display rich dynamics, going from staying constant to disappearing and reappearing in specific cluster domains within minutes.

AB - Sub-diffraction imaging of plasma membrane localized proteins, such as the SNARE (Soluble NSF Attachment Protein Receptor) proteins involved in exocytosis, in fixed cells have resulted in images with high spatial resolution, at the expense of dynamical information. Here, we have imaged localized fluorescence bursts of DRONPA-fused SNAP-25 molecules in live chromaffin cells by Total Internal Reflection Fluorescence (TIRF) imaging. We find that this method allows tracking protein cluster dynamics over relatively long times (∼20 min.), partly due to the diffusion into the TIRF field of fresh molecules, making possible the simultaneous identification of cluster size, location and temporal evolution. The results indicate that the DRONPA-fused SNAP-25 clusters display rich dynamics, going from staying constant to disappearing and reappearing in specific cluster domains within minutes.

U2 - 10.1039/c4pp00423j

DO - 10.1039/c4pp00423j

M3 - Journal article

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SN - 1474-905X

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EP - 1012

JO - Photochemical & Photobiological Sciences

JF - Photochemical & Photobiological Sciences

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Spatial distribution and temporal evolution of DRONPA-fused SNAP25 clusters in adrenal chromaffin cells

Abstract

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