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Imaging dynamic cell-cell junctional coupling in vivo using Trojan-LAMP
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Imaging dynamic cell-cell junctional coupling in vivo using Trojan-LAMP
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Imaging dynamic cell-cell junctional coupling in vivo using Trojan-LAMP
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Journal Article
Imaging dynamic cell-cell junctional coupling in vivo using Trojan-LAMP
2008
Overview
Cell-cell coupling via gap junctions has been extensively studied
in vitro
and in heterologous systems, but
in vivo
studies are still few. A new class of photoactivatable bioconjugates is now used to monitor gap junctional coupling in living
Caenorhabditis elegans
.
To study the physiological regulation and function of cell-cell gap junction communication
in vivo
, we developed a bioconjugate of caged dye, named dextran-CANPE-HCC, for imaging cell coupling in small model organisms.
In vitro
, the compound was photolyzed efficiently with robust fluorescence enhancement. Dextran-CANPE-HCC delivered into
Caenorhabditis elegans
oocytes was retained in cells throughout development. Using local uncaging, we photolyzed dextran-CANPE-HCC to release the small HCC dye and imaged the dynamics of intercellular dye transfer through gap junction channels, a technique we named Trojan–local activation of molecular fluorescent probes (LAMP). Early during embryonic development, the pattern of cell coupling undergoes dramatic remodeling and imaging revealed that the germ cell precursors, P2, P3 and P4, were isolated from the somatic cell communication compartment. As dextran-CANPE-HCC is chemically and metabolically stable, labeled worms showed very bright signal upon photoactivation after hatching, which allowed us to examine cell coupling in living worms noninvasively.
Publisher
Nature Publishing Group US,Nature Publishing Group
