A micropatterning approach for imaging dynamic Cx43 trafficking to cell-cell borders

Shan Shan Zhang; Soongweon Hong; André G. Kléber; Luke P. Lee; Robin M. Shaw

doi:10.1016/j.febslet.2014.01.002

A micropatterning approach for imaging dynamic Cx43 trafficking to cell-cell borders

Shan Shan Zhang, Soongweon Hong, André G. Kléber, Luke P. Lee, Robin M. Shaw

Chemistry & Nanoscience

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

Abstract

The precise expression and timely delivery of connexin 43 (Cx43) proteins to form gap junctions are essential for electrical coupling of cardiomyocytes. Growing evidence supports a cytoskeletal-based trafficking paradigm for Cx43 delivery directly to adherens junctions at the intercalated disc. A limitation of Cx43 localization assays in cultured cells, in which cell-cell contacts are essential, is the inability to control for cell geometry or reproducibly generate contact points. Here we present a micropatterned cell pairing system well suited for live microscopy to examine how the microtubule and actin cytoskeleton confer specificity to Cx43 trafficking to precisely defined cell-cell junctions. This system can be adapted for other cell types and used to study dynamic intracellular movements of other proteins important for cell-cell communication.

Original language	English
Pages (from-to)	1439-1445
Number of pages	7
Journal	FEBS Letters
Volume	588
Issue number	8
DOIs	https://doi.org/10.1016/j.febslet.2014.01.002
State	Published - 17 Apr 2014

Keywords

Connexin 43
Cytoskeleton
Micropatterning
Trafficking

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10.1016/j.febslet.2014.01.002

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title = "A micropatterning approach for imaging dynamic Cx43 trafficking to cell-cell borders",

abstract = "The precise expression and timely delivery of connexin 43 (Cx43) proteins to form gap junctions are essential for electrical coupling of cardiomyocytes. Growing evidence supports a cytoskeletal-based trafficking paradigm for Cx43 delivery directly to adherens junctions at the intercalated disc. A limitation of Cx43 localization assays in cultured cells, in which cell-cell contacts are essential, is the inability to control for cell geometry or reproducibly generate contact points. Here we present a micropatterned cell pairing system well suited for live microscopy to examine how the microtubule and actin cytoskeleton confer specificity to Cx43 trafficking to precisely defined cell-cell junctions. This system can be adapted for other cell types and used to study dynamic intracellular movements of other proteins important for cell-cell communication.",

keywords = "Connexin 43, Cytoskeleton, Micropatterning, Trafficking",

author = "Zhang, {Shan Shan} and Soongweon Hong and Kl{\'e}ber, {Andr{\'e} G.} and Lee, {Luke P.} and Shaw, {Robin M.}",

note = "Funding Information: We thank Roger Chang for setting up the single-particle tracking software in Matlab, and Dr. Priscilla J. Chan for work on micropattern development. We also thank Drs. James W. Smyth and Matthew L. Wheeler for critical review of this manuscript. SS Zhang and RM Shaw are supported by the American Heart Association (Postdoctoral and Established Investigator Awards), as well as by the NIH/NHLBI (RO1). Appendix A ",

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doi = "10.1016/j.febslet.2014.01.002",

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T1 - A micropatterning approach for imaging dynamic Cx43 trafficking to cell-cell borders

AU - Zhang, Shan Shan

AU - Hong, Soongweon

AU - Kléber, André G.

AU - Lee, Luke P.

AU - Shaw, Robin M.

N1 - Funding Information: We thank Roger Chang for setting up the single-particle tracking software in Matlab, and Dr. Priscilla J. Chan for work on micropattern development. We also thank Drs. James W. Smyth and Matthew L. Wheeler for critical review of this manuscript. SS Zhang and RM Shaw are supported by the American Heart Association (Postdoctoral and Established Investigator Awards), as well as by the NIH/NHLBI (RO1). Appendix A

PY - 2014/4/17

Y1 - 2014/4/17

N2 - The precise expression and timely delivery of connexin 43 (Cx43) proteins to form gap junctions are essential for electrical coupling of cardiomyocytes. Growing evidence supports a cytoskeletal-based trafficking paradigm for Cx43 delivery directly to adherens junctions at the intercalated disc. A limitation of Cx43 localization assays in cultured cells, in which cell-cell contacts are essential, is the inability to control for cell geometry or reproducibly generate contact points. Here we present a micropatterned cell pairing system well suited for live microscopy to examine how the microtubule and actin cytoskeleton confer specificity to Cx43 trafficking to precisely defined cell-cell junctions. This system can be adapted for other cell types and used to study dynamic intracellular movements of other proteins important for cell-cell communication.

AB - The precise expression and timely delivery of connexin 43 (Cx43) proteins to form gap junctions are essential for electrical coupling of cardiomyocytes. Growing evidence supports a cytoskeletal-based trafficking paradigm for Cx43 delivery directly to adherens junctions at the intercalated disc. A limitation of Cx43 localization assays in cultured cells, in which cell-cell contacts are essential, is the inability to control for cell geometry or reproducibly generate contact points. Here we present a micropatterned cell pairing system well suited for live microscopy to examine how the microtubule and actin cytoskeleton confer specificity to Cx43 trafficking to precisely defined cell-cell junctions. This system can be adapted for other cell types and used to study dynamic intracellular movements of other proteins important for cell-cell communication.

KW - Connexin 43

KW - Cytoskeleton

KW - Micropatterning

KW - Trafficking

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EP - 1445

JO - FEBS Letters

JF - FEBS Letters

IS - 8

ER -

A micropatterning approach for imaging dynamic Cx43 trafficking to cell-cell borders

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Keywords

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