Experimental control of excitable embryonic tissues: three stimuli induce rapid epithelial contraction.
Cell generated contractility is a major driver of morphogenesis during processes such as epithelial bending and epithelial-to-mesenchymal transitions. Previous studies of contraction in embryos have relied on developmentally programmed cell shape changes such as those that accompany ventral furrow formation in Drosophila, bottle cell formation in Xenopus, ingression in amniote embryos, and neurulation in vertebrate embryos. We have identified three methods to reproducibly and acutely induce contraction in embryonic epithelial sheets: laser activation, electrical stimulation, and nano-perfusion with chemicals released by wounding. Contractions induced by all three methods occur over a similar time-scale (1 to 2 min) and lead to reorganization of the F-actin cytoskeleton. By combining induced contractions with micro-aspiration we can simultaneously measure the stiffness of the tissue and the force and work done by contractions. Laser activation allows real-time visualization of F-actin remodeling during contraction. Perfusion with cell lysate suggests that these three stimuli activate physiologically relevant pathways that maintain epithelial tension or trigger epithelial morphogenesis. Our methods provide the means to control and study cellular contractility and will allow dissection of molecular mechanisms and biomechanics of cellular contractility.
Joshi
Sagar D
SD
Department of Bioengineering, University of Pittsburgh, 3501 Fifth Ave., Pittsburgh, PA 15260, USA.
von Dassow
Michelangelo
M
Davidson
Lance A
LA
D008841:Actin Cytoskeleton / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D000465:Algorithms; D000818:Animals; D001696:Biomechanical Phenomena; D002457:Cell Extracts / Q000494:pharmacology; D003599:Cytoskeleton / Q000378:metabolism; D004548:Elasticity; D004558:Electric Stimulation; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000502:physiology* / Q000528:radiation effects; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology* / Q000528:radiation effects; D004848:Epithelium / Q000187:drug effects / Q000196:embryology* / Q000528:radiation effects; D053685:Laser Therapy; D008853:Microscopy; D009024:Morphogenesis / Q000502:physiology; D010477:Perfusion; D014982:Xenopus laevis
D000199:Actins; D002457:Cell Extracts
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