Fluorescent Labeling of Connexin with As Complex and X-Y Coordinate Registration of Target Single Cells Based on a Triangle Standard Chip for the Image Analysis of Gap Junctional Communication

2020 ◽  
Author(s):  
Mikako Saito
Keyword(s):  
Image Analysis ◽  
Single Cells ◽  
Fluorescent Labeling ◽  
Gap Junctional Communication ◽  
Junctional Communication ◽  
Standard Chip ◽  
Gap Junctional

scholarly journals Asymmetric patterns of gap junctional communication in developing chicken skin

Development ◽  
1993 ◽  
Vol 119 (1) ◽  
pp. 85-96 ◽  
Author(s):  
F. Serras ◽  
S. Fraser ◽  
C.M. Chuong
Keyword(s):  
Lucifer Yellow ◽  
Single Cells ◽  
Dye Coupling ◽  
Dye Transfer ◽  
Gap Junctional Communication ◽  
Junctional Communication ◽  
Chicken Skin ◽  
Preferential Pathways ◽  
Gap Junctional ◽  
Feather Development

To study the pattern of gap junctional communication in chicken skin and feather development, we injected Lucifer Yellow into single cells and monitored the transfer of the fluorescent dye through gap junctions. Dye coupling is present between cells of the epithelium as well as between cells of the mesoderm. However, dye transfer did not occur equally in all directions and showed several consistent patterns and asymmetries, including: (1) no dye coupling between mesoderm and epithelium, (2) partial restriction of dye coupling at the feather bud/interbud boundary during early feather bud development, (3) preferential distribution of Lucifer Yellow along the anteroposterior axis of the feather placode and (4) absence of dye coupling in some epithelial cells. These results suggest the presence of preferential pathways of communication that may play a role in the patterning of chicken skin.

scholarly journals Inhibition of Endothelial Wound Repair by Dominant Negative Connexin Inhibitors

2001 ◽  
Vol 12 (4) ◽  
pp. 831-845 ◽  
Author(s):  
Brenda R. Kwak ◽  
Michael S. Pepper ◽  
Daniel B. Gros ◽  
Paolo Meda
Keyword(s):  
Endothelial Cells ◽  
Wound Repair ◽  
Single Cells ◽  
Dominant Negative ◽  
Cell Coupling ◽  
Mechanical Wounding ◽  
Functional Changes ◽  
Gap Junctional Communication ◽  
Junctional Communication ◽  
Gap Junctional

Wounding of endothelial cells is associated with altered direct intercellular communication. To determine whether gap junctional communication participates to the wound repair process, we have compared connexin (Cx) expression, cell-to-cell coupling and kinetics of wound repair in monolayer cultures of PymT-transformed mouse endothelial cells (clone bEnd.3) and in bEnd.3 cells expressing different dominant negative Cx inhibitors. In parental bEnd.3 cells, mechanical wounding increased expression of Cx43 and decreased expression of Cx37 at the site of injury, whereas Cx40 expression was unaffected. These wound-induced changes in Cx expression were associated with functional changes in cell-to-cell coupling, as assessed with different fluorescent tracers. Stable transfection with cDNAs encoding for the chimeric connexin 3243H7 or the fusion protein Cx43-βGal resulted in perturbed gap junctional communication between bEnd.3 cells under both basal and wounded conditions. The time required for complete repair of a defined wound within a confluent monolayer was increased by ∼50% in cells expressing the dominant negative Cx inhibitors, whereas other cell properties, such as proliferation rate, migration of single cells, cyst formation and extracellular proteolytic activity, were unaltered. These findings demonstrate that proper Cx expression is required for coordinated migration during repair of an endothelial wound.

Reduced junctional permeability at interrhombomeric boundaries

Development ◽  
1992 ◽  
Vol 116 (4) ◽  
pp. 1069-1076 ◽  
Author(s):  
S. Martinez ◽  
E. Geijo ◽  
M.V. Sanchez-Vives ◽  
L. Puelles ◽  
R. Gallego
Keyword(s):  
Lucifer Yellow ◽  
Single Cells ◽  
Electrical Coupling ◽  
Dye Transfer ◽  
Gap Junctional Communication ◽  
Junctional Communication ◽  
Neuroepithelial Cells ◽  
Junctional Permeability ◽  
Pattern Specification ◽  
Gap Junctional

Intercellular communication is considered to have a role during pattern specification processes in early embryonic development. This report analyzes the changing gap junctional communication properties of chick neuroepithelial cells depending on their position relative to the segmental partitions of the rhombencephalon. Intercellular electrical coupling and dye transfer were studied with microelectrode techniques. Neuroepithelial cells were electrically coupled irrespective of their location relative to interneuromeric boundaries. Iontophoretic injection of biocytin or Lucifer Yellow into single cells inside the rhombomeres was followed by transjunctional diffusion to the surrounding cells. In contrast, dye transfer was strictly limited when the diffusion zone contacted the cells forming the interneuromeric limits. Label injected into the boundary cells did not spread to other cells at all. Avian interrhombomeric boundaries are thus sites of reduced junctional permeability during early morphogenesis.

scholarly journals Disruption of Gap Junctional Communication by the Platelet-derived Growth Factor Is Mediated via Multiple Signaling Pathways

1999 ◽  
Vol 274 (15) ◽  
pp. 10489-10496 ◽  
Author(s):  
Mohammad Z. Hossain ◽  
Ajit B. Jagdale ◽  
Peng Ao ◽  
Andrius Kazlauskas ◽  
Alton L. Boynton
Keyword(s):  
Growth Factor ◽  
Signaling Pathways ◽  
Platelet Derived Growth Factor ◽  
Gap Junctional Communication ◽  
Junctional Communication ◽  
Gap Junctional ◽  
Multiple Signaling

Role of protein kinase C in the regulation of gap junctional communication

1994 ◽  
Vol 14 (6) ◽  
pp. 259-270 ◽  
Author(s):  
Irina V. Budunova ◽  
Leonid A. Mittelman ◽  
Joanna Miloszewska
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Gap Junctional Communication ◽  
Kinase C ◽  
Junctional Communication ◽  
Gap Junctional

Gap junction-mediated transfer of left-right patterning signals in the early chick blastoderm is upstream of Shh asymmetry in the node

Development ◽  
1999 ◽  
Vol 126 (21) ◽  
pp. 4703-4714 ◽  
Author(s):  
M. Levin ◽  
M. Mercola
Keyword(s):  
Gene Expression ◽  
Gap Junction ◽  
Large Scale ◽  
Chick Blastoderm ◽  
Gap Junction Channels ◽  
Gap Junctional Communication ◽  
Junctional Communication ◽  
Left And Right ◽  
The Right ◽  
Gap Junctional

Invariant patterning of left-right asymmetry during embryogenesis depends upon a cascade of inductive and repressive interactions between asymmetrically expressed genes. Different cascades of asymmetric genes distinguish the left and right sides of the embryo and are maintained by a midline barrier. As such, the left and right sides of an embryo can be viewed as distinct and autonomous fields. Here we describe a series of experiments that indicate that the initiation of these programs requires communication between the two sides of the blastoderm. When deprived of either the left or the right lateral halves of the blastoderm, embryos are incapable of patterning normal left-right gene expression at Hensen's node. Not only are both flanks required, suggesting that there is no single signaling source for LR pattern, but the blastoderm must be intact. These results are consistent with our previously proposed model in which the orientation of LR asymmetry in the frog, Xenopus laevis, depends on large-scale partitioning of LR determinants through intercellular gap junction channels (M. Levin and M. Mercola (1998) Developmental Biology 203, 90–105). Here we evaluate whether gap junctional communication is required for the LR asymmetry in the chick, where it is possible to order early events relative to the well-characterized left and right hierarchies of gene expression. Treatment of cultured chick embryos with lindane, which diminishes gap junctional communication, frequently unbiased normal LR asymmetry of Shh and Nodal gene expression, causing the normally left-sided program to be recapitulated symmetrically on the right side of the embryo. A survey of early expression of connexin mRNAs revealed that Cx43 is present throughout the blastoderm at Hamburger-Hamilton stage 2–3, prior to known asymmetric gene expression. Application of antisense oligodeoxynucleotides or blocking antibody to cultured embryos also resulted in bilateral expression of Shh and Nodal transcripts. Importantly, the node and primitive streak at these stages lack Cx43 mRNA. This result, together with the requirement for an intact blastoderm, suggests that the path of communication through gap junction channels circumvents the node and streak. We propose that left-right information is transferred unidirectionally throughout the epiblast by gap junction channels in order to pattern left-sided Shh expression at Hensen's node.

Syncytial organization of cultured rat mesangial cells

1991 ◽  
Vol 260 (6) ◽  
pp. F848-F855 ◽  
Author(s):  
K. Iijima ◽  
L. C. Moore ◽  
M. S. Goligorsky
Keyword(s):  
Calcium Channels ◽  
Mesangial Cells ◽  
Calcium Waves ◽  
Oxygen Intermediates ◽  
Voltage Gated ◽  
Fura 2 ◽  
Gap Junctional Communication ◽  
Voltage Gated Calcium Channels ◽  
Junctional Communication ◽  
Gap Junctional

To investigate communication competence of cultured rat mesangial cells, Lucifer yellow transfer was studied using microinjection and scrape-loading techniques. Both methods yielded results indicating considerable gap junctional communication between cultured mesangial cells. Gap junctional communication between mesangial cells was upregulated by adenosine 3',5'-cyclic monophosphate (cAMP). Conversely, cell-to-cell communication was attenuated by exposure to the tumor promoter phorbol myristate acetate, the Ca ionophore ionomycin, reduced oxygen intermediates, and cell acidification. Expression of voltage gated calcium channels by mesangial cells was studied microspectrofluorimetrically using fura-2 fluorescence. KCl-induced depolarization, BAY-K 8644, and readdition of calcium to Ca-free depolarizing medium all produced a nifedipine-inhibitable increase in cytosolic calcium concentration. The existence of voltage-gated calcium channels in communication-competent cells suggests the possibility of propagation of depolarizing signals across the syncytium. This was studied by microapplication of KCl to the microenvironment of a single cell and monitoring fura-2 fluorescence in remote cells. This maneuver resulted in propagating calcium waves in communication-competent monolayers; calcium waves could not be evoked in monolayers exposed to an alkanol-type gap junction uncoupler, octanol. It is concluded that cultured rat mesangial cells form a syncytium capable of propagating calcium transients from a single depolarized cell to its coupled neighbors.

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