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*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:24046*  Cite 
Description

Starved Dictyostelium cells lacking both guanylyl cyclases (GCase A and GCase), but expressing GCase with an inactivated catalytic domain (sGCΔCat) and in medium supplemented with 60 µM LY294002 (a PI3K inhibitor) and 1µM cAMP migrate continuously toward the anode (right). A direct-current electric field of 10 Vpcm was applied and the cells were observed for 20 min at 5-sec intervals. (Scale bar, 50 µm.) In contrast, cathode-directed migration is strongly attenuated in wild-type cells in the same medium. Compare with wild-type control video (CIL# 24043) and other mutants lacking guanylyl cyclase activity (CIL#24044, 24045). Videos are supporting information in Proc Natl Acad Sci (2009). 106: 6667-6672.

Technical Details

Starved cells were suspended in buffer containing 10 mM Na/K PO4, 2 mM MgSO4, and 0.2 mM CaCl2, (pH 6.5) and 4 mM caffeine to inhibit adenylyl cyclase activity (which also reduced cell–cell interactions). A chamber was constructed to allow application of a direct-current electric field (Sato et al. 2007. BioSystems). The cells in the chamber were observed with an Olympus IX-71 inverted microscope with phase contrast optics. Cell behavior was recorded with a cooled CCD camera (MicroMax; Princeton Instruments) and software (MetaMorph; Molecular Devices).

Biological Sources
NCBI Organism Classification
Dictyostelium discoideum AX2
Cell Type
amoeboid cell
Cell Line
AX2-gc-null/sGCdeltaCat
Cellular Component
cytosol
intracellular
Attribution
Names
Masayuki J. Sato
Hidekazu Kuwayama
Wouter N. van Egmond
Airi L. K. Takayama
Hiroaki Takagi
Peter J. M. van Haastert
Toshio Yanagida
Masahiro Ueda
Published
Proc Natl Acad Sci (2009). 106: 6667-6672.
Pubmed
19346484
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL24046
Archival Resource Key (ARK)
ark:/b7295/w9cil24046
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
phase contrast microscopy
time lapse microscopy
Parameters Imaged
elastic scattering of photons
Source of Contrast
differences in amount of elastic light scattering
Processing History
unprocessed raw data
Data Qualifiers
raw, unprocessed data
Sample Preparation
Methods
living tissue
Relation To Intact Cell
whole mounted tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 452px ——
Y 240px ——
Time 5 seconds 242