The effect of complete carcinogens on intercellular transfer of lucifer yellow in fibroblast culture

1990 ◽  
Vol 6 (1) ◽  
pp. 47-61 ◽  
Author(s):  
Irene V. Budunova ◽  
Leonid A. Mittelman ◽  
Gennady A. Belitsky
Keyword(s):  
Lucifer Yellow ◽  
Fibroblast Culture ◽  
Intercellular Transfer ◽  
Complete Carcinogens

scholarly journals Transfected connexin45 alters gap junction permeability in cells expressing endogenous connexin43.

1995 ◽  
Vol 130 (4) ◽  
pp. 987-995 ◽  
Author(s):  
M Koval ◽  
S T Geist ◽  
E M Westphale ◽  
A E Kemendy ◽  
R Civitelli ◽  
...  
Keyword(s):  
Gap Junction ◽  
Lucifer Yellow ◽  
Direct Interaction ◽  
Full Length ◽  
Osteosarcoma Cell ◽  
Dye Transfer ◽  
Fluorescent Reporter ◽  
Intercellular Transfer ◽  
Carboxyl Terminal ◽  
Gap Junctional

Many cells express multiple connexins, the gap junction proteins that interconnect the cytosol of adjacent cells. Connexin43 (Cx43) channels allow intercellular transfer of Lucifer Yellow (LY, MW = 443 D), while connexin45 (Cx45) channels do not. We transfected full-length or truncated chicken Cx45 into a rat osteosarcoma cell line ROS-17/2.8, which expresses endogenous Cx43. Both forms of Cx45 were expressed at high levels and colocalized with Cx43 at plasma membrane junctions. Cells transfected with full-length Cx45 (ROS/Cx45) and cells transfected with Cx45 missing the 37 carboxyl-terminal amino acids (ROS/Cx45tr) showed 30-60% of the gap junctional conductance exhibited by ROS cells. Intercellular transfer of three negatively charged fluorescent reporter molecules was examined. In ROS cells, microinjected LY was transferred to an average of 11.2 cells/injected cell, while dye transfer between ROS/Cx45 cells was reduced to 3.9 transfer between ROS/Cx45 cells was reduced to 3.9 cells. In contrast, ROS/Cx45tr cells transferred LY to > 20 cells. Transfer of calcein (MW = 623 D) was also reduced by approximately 50% in ROS/Cx45 cells, but passage of hydroxycoumarin carboxylic acid (HCCA; MW = 206 D) was only reduced by 35% as compared to ROS cells. Thus, introduction of Cx45 altered intercellular coupling between cells expressing Cx43, most likely the result of direct interaction between Cx43 and Cx45. Transfection of Cx45tr and Cx45 had different effects in ROS cells, consistent with a role of the carboxyl-terminal domain of Cx45 in determining gap junction permeability or interactions between connexins. These data suggest that coexpression of multiple connexins may enable cells to achieve forms of intercellular communication that cannot be attained by expression of a single connexin.

Identification of tumor promoters by their inhibitory effect on intercellular transfer of lucifer yellow

1989 ◽  
Vol 5 (1) ◽  
pp. 77-89 ◽  
Author(s):  
Irene V. Budunova ◽  
Leonid A. Mittelman ◽  
Gennady A. Belitsky
Keyword(s):  
Lucifer Yellow ◽  
Inhibitory Effect ◽  
Tumor Promoters ◽  
Intercellular Transfer

Multimodal Atomic Force Imaging of Open Hemichannelinduced Modulation of Cell Volume and Viscoelastic Properties

1999 ◽  
Vol 5 (S2) ◽  
pp. 998-999
Author(s):  
Seung K. Rhee ◽  
Arjan P. Quist ◽  
Hai Lin ◽  
Nils Almqvist ◽  
Ratneshx Lai
Keyword(s):  
Plasma Membrane ◽  
Cell Volume ◽  
Lucifer Yellow ◽  
Single Cells ◽  
Dye Uptake ◽  
Aortic Endothelial Cells ◽  
Atomic Force ◽  
Cell Plasma ◽  
Uptake Assay ◽  
Gap Junctional

Hemichannels from two single cells can join upon contact between these cells to form gap junctions - an intercellular pathway for the direct exchange of ions and small metabolites. Using techniques of fluorescent dye-uptake assay, laser confocal fluorescence imaging and atomic force microscopy (AFM), we have examined the role of hemichannels, present in the non-junctional regions of single cell plasma membrane, in the modulation of cell volume.Antibodies against a gap junctional protein connexin43, were immunolocalized to nonjunctional plasma membrane regions of single BICR-MlRk k (breast tumor epithelial) cells, KOM-1 (bovine aortic endothelial) cells, and GM04260 (AD-free human) fibroblast cells. In the absence of extracellular calcium, cytoplasmic uptake of Lucifer yellow (LY) but not of dextran-conjugated LY was observed in single cells. Dye uptake was prevented by gap junctional inhibitors, ẞ-glycyrrhetinic acid (ẞGCA) and oleamide.

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