A micropatterning approach for imaging dynamic Cx43 trafficking to cell-cell borders.
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.
Cedars-Sinai Heart Institute, Los Angeles, CA 90048, USA.
Shan-Shan
SS
Zhang
Departments of Electrical Engineering and Computer Science and Biophysics Graduate Group, University of California, Berkeley, CA 94720, USA.
SoonGweon
S
Hong
Department of Pathology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 02215, USA.
André G
AG
Kléber
Departments of Electrical Engineering and Computer Science and Biophysics Graduate Group, University of California, Berkeley, CA 94720, USA.
Luke P
LP
Lee
Cedars-Sinai Heart Institute, Los Angeles, CA 90048, USA; Department of Medicine, University of California, Los Angeles, CA 90095, USA. Electronic address: Robin.Shaw@cshs.org.
Robin M
RM
Shaw
D000818:Animals; D002478:Cells, Cultured; D018031:Connexin 43 / Q000378:metabolism*; D017629:Gap Junctions / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence / Q000379:methods; D032383:Myocytes, Cardiac / Q000378:metabolism; D061848:Optical Imaging / Q000379:methods; D021381:Protein Transport
D018031:Connexin 43
Connexin 43; Cytoskeleton; Micropatterning; Trafficking