Dye-coupling between term pregnant human myometrial cells before labor: carboxyfluorescein versus lucifer yellow

1995 ◽  
Vol 19 (7) ◽  
pp. 609-618 ◽  
Author(s):  
H Çiray
Keyword(s):  
Lucifer Yellow ◽  
Dye Coupling ◽  
Myometrial Cells

scholarly journals Neuropeptides Exert Direct Effects on Rat Thymic Epithelial Cells in Culture

1998 ◽  
Vol 6 (1-2) ◽  
pp. 95-104 ◽  
Author(s):  
Gail M. Head ◽  
R. Mentlein ◽  
Birte Von Patay ◽  
J. E.G. Downing ◽  
Marion D. Kendall
Keyword(s):  
Epithelial Cells ◽  
Lucifer Yellow ◽  
Tritiated Thymidine ◽  
Single Cells ◽  
Thymic Epithelial Cells ◽  
Dye Coupling ◽  
Direct Effects ◽  
Adrenergic Agonist ◽  
Thymic Epithelium ◽  
Functional Aspects

To determine if major thymic neuropeptides and neurotransmitters can directly influence the functional activity of cultured rat thymic epithelium, neuropeptides and neurotransmitters were applied, and intercellular communication, proliferation, and thymulin secretion assessed. After injections of a mixture of lucifer yellow dextran (too large to pass gap junctions) and cascade blue (which does) into single cells, some neuropeptides decrease dye coupling: 0.1 mM GABA (P< 0.0001), 100 nM NPY (P< 0.0001), 100 nM VIP (P< 0.001), 100 nM CGRP (P< 0.001), 100 nM SP (P< 0.01), and 0.1 mM histamine (P< 0.01), whereas 0.1 mM 5-HT, mM acetylcholine, and 1μM isoproterenol (β-adrenergic agonist) had no effect. Proliferation (incorporation of tritiated thymidine) was increased by CGRP (P= 0.004) and histamine (P< 0.02), but decreased by isoproterenol (P= 0.002), 5-HT (P= 0.003), and acetylcholine (P< 0.05). The percentage of multinucleate cells was decreased after isoproterenol (2.5%), and increased after 5-HT (21.3%), GABA (15%), and histamine (15.1%). Compared to controls, thymulin in the supernatant was decreased after challenge with acetylcholine (52%), isoproterenol (71%), 5-HT (73%), and histamine (84%). This study demonstrates direct effects of neuropeptides and neurotransmitters on functional aspects of cultured thymic epithelial cells.

Relationship between Na+ current expression and cell-cell coupling in astrocytes cultured from rat hippocampus

1991 ◽  
Vol 65 (4) ◽  
pp. 989-1002 ◽  
Author(s):  
H. Sontheimer ◽  
S. G. Waxman ◽  
B. R. Ransom
Keyword(s):  
Lucifer Yellow ◽  
Na Channel ◽  
Dye Coupling ◽  
Cell Coupling ◽  
Na Current ◽  
Na Currents ◽  
Coupled Cells ◽  
Hippocampal Astrocytes ◽  
Cell Cell

1. Cell-cell coupling between hippocampal astrocytes in culture was studied by following the intracellular spread of the low molecular weight fluorescent dye Lucifer yellow (LY). Dye coupling appeared as early as 24 h after plating, at which time approximately 20% of all astrocytes that physically contacted neighboring cells showed dye coupling. 2. The percentage of coupled cells increased with time in culture and peaked after 10 days in vitro (DIV) when approximately 50% of astrocytes showed coupling. Further time in culture, up to 20 DIV, did not increase the percentage of coupled cells. Thus, coupled and noncoupled astrocytes coexist in hippocampal cultures in approximately equal numbers. 3. Na+ currents were expressed in a subpopulation of hippocampal astrocytes and changed characteristics during in vitro development. A "neuronal type" of Na+ current, so called because of an h alpha curve that had a midpoint near -60 mV, was observed within the first 5 days post-plating. A "glial type" of Na+ current, characterized by a -25 mV shift in its h alpha curve, was only expressed after 6 days in culture. 4. Na+ current expression was restricted to hippocampal astrocytes that did not exhibit dye coupling; astrocytes that exhibited dye coupling (n = 39) did not show measurable Na+ currents. 5. The failure to see Na+ currents in coupled astrocytes cannot be explained by insufficient space-clamp since astrocytes acutely uncoupled with octanol (10 microM) did not reveal Na+ current expression. Control experiments showed that low concentrations of octanol (i.e., 10-100 microM) did not block Na+ currents; blockage of Na+ currents by octanol was only observed at high concentrations (e.g., 50-fold the concentration used for uncoupling). These observations support the idea that Na(+)-channel expression was restricted to noncoupled astrocytes. 6. The time courses for the development of cell coupling and Na+ current expression appeared to be inversely correlated and suggested a gradual increase in cell coupling in concert with a loss in Na+ current expression with time in culture.

Reversible carbon dioxide-induced inhibition of dye coupling in Necturus gallbladder

1983 ◽  
Vol 244 (5) ◽  
pp. C419-C421 ◽  
Author(s):  
J. A. Jarrell
Keyword(s):  
Carbon Dioxide ◽  
Lucifer Yellow ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Dye Coupling ◽  
Gallbladder Epithelium ◽  
Intracellular Injection ◽  
Necturus Gallbladder ◽  
High Concentrations

The cells of Necturus gallbladder epithelium are electrically coupled. This work used intracellular injection of the fluorescent dye Lucifer yellow to demonstrate that these cells are also dye coupled and that this coupling is rapidly and reversibly inhibited by high concentrations of carbon dioxide. Dye coupling is also inhibited by the calcium ionophore A23187.

Gap junctions in human umbilical cord endothelial cells contain multiple connexins

1997 ◽  
Vol 272 (1) ◽  
pp. C117-C130 ◽  
Author(s):  
H. Van Rijen ◽  
M. J. van Kempen ◽  
L. J. Analbers ◽  
M. B. Rook ◽  
A. C. van Ginneken ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Lucifer Yellow ◽  
Umbilical Vein ◽  
Electrical Conductance ◽  
Distribution Patterns ◽  
Human Umbilical Cord ◽  
Dye Coupling ◽  
Gap Junctional

We investigated the expression pattern of gap junctional proteins (connexins, Cx) in situ and in vitro and their functional characteristics in cultured human umbilical vein endothelial cells (HUVEC) and cultured human umbilical artery endothelial cells (HUAEC). In both arteries and veins, Cx37, Cx40, and Cx43 could be detected in situ and in vitro (passages 2-4). Distribution patterns of Cx40 and Cx43 were homogeneous in situ but more heterogeneous in vitro. Cx37 is heterogeneously expressed both in situ and in vitro. Among most cells, no Cx37 staining could be detected; when present, it was found as bright spots between some clusters of cells. Cx40 was more abundant in cultured arterial endothelium than in cultured venous endothelium. Dye-coupling experiments with Lucifer yellow CH revealed extensive dye spread in HUVEC (15.2 +/- 0.4, mean +/- SE, n = 110) but was significantly restricted in HUAEC (9.8 +/- 0.3, n = 110). Electrophysiological gap junctional characteristics were determined in cultured HUVEC and HUAEC pairs by use of the dual voltage-clamp technique. In contrast to the dye-coupling experiments, mean macroscopic electrical conductance was significantly larger for HUAEC pairs (31.4 +/- 6.0 nS, n = 12) than for HUVEC pairs (16.6 +/- 2.8, n = 18). In HUVEC, we measured multiple single gap junctional channel conductances in the range of 19-75 pS. Interestingly, additional conductances of 80-200 pS were measured in HUAEC, possibly partially reflecting activity of channels formed of Cx40, which are more abundant in the cultured arterial endothelial cells.

scholarly journals Gap junctional communication in the extraembryonic tissues of the gastrulating mouse embryo.

1989 ◽  
Vol 109 (6) ◽  
pp. 3015-3026 ◽  
Author(s):  
G H Kalimi ◽  
C W Lo
Keyword(s):  
Mouse Embryo ◽  
Lucifer Yellow ◽  
Dye Coupling ◽  
Gap Junctional Communication ◽  
Stage Of Development ◽  
Junctional Communication ◽  
Gap Junctional ◽  
Ectoplacental Cone ◽  
Ionic Coupling ◽  
Extraembryonic Tissues

We characterized gap junctional communication in the extraembryonic tissues of the 7.5-d gastrulating mouse embryo. At this stage of development, the extraembryonic tissues form a large part of the conceptus, and link the embryo proper to the maternal tissue. Using Lucifer yellow injections, cells in most extraembryonic tissues were observed to be very well dye coupled, the only exception being the peripheral regions of the ectoplacental cone. Of particular interest was the fact that no dye coupling was detected between the three major extraembryonic tissues. Thus, the extraembryonic ectoderm (EEC), the extraembryonic endoderm (EEN), and the ectoplacental cone (EPC) corresponded to separate communication compartments, with the EPC being further subdivided into three compartments. Interestingly, the EEN was observed to exhibit a very low level of dye coupling with the adjacent visceral embryonic endoderm (EN), and consistent with the latter dye coupling results was the finding that the EEN was ionically coupled to the EN, but not with any other extraembryonic tissues. However, in the EPC, ionic coupling studies show that the central region was well coupled ionically to the EEC, but only weakly coupled to the peripheral EPC. These findings, in conjunction with our previous study (1988. J. Cell Biol. 107:241-255), demonstrate that the 7.5-d mouse conceptus is subdivided into at least nine major Lucifer yellow-delineated communication compartments, with ionic coupling across some of these compartments effectively unifying the embryo into two large domains corresponding to the embryo proper and the major extraembryonic tissues.

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 Dye coupling in the muscles controlling squid chromatophore expansion.

1995 ◽  
Vol 198 (12) ◽  
pp. 2631-2634 ◽  
Author(s):  
C M Reed
Keyword(s):  
Sea Water ◽  
Lucifer Yellow ◽  
Muscle Fibres ◽  
Single Muscle ◽  
Dye Coupling ◽  
Living Animal ◽  
Dye Transfer ◽  
High Concentrations ◽  
Radial Muscle

Dye coupling between the cone-shaped radial muscle fibres, which control the expansion and closing of a squid chromatophore organ, was investigated in the squid Loligo vulgaris. Particular attention was paid to the role of the myomuscular junctions located between the muscle fibres. Lucifer Yellow was injected ionophoretically into single muscle fibres under normal artificial sea water (ASW) and under various concentrations of calcium in ASW. Under ASW, 44% of muscle fibres examined were dye-coupled, 82% were coupled under calcium-free sea water and 67% were coupled under sea water containing high concentrations of calcium. Dye transfer was blocked by octanol. Muscle fibres were never seen to link adjacent chromatophore organs. Results are discussed in terms of the role of the myomuscular junctions in the regulation of chromatophore expansion in the living animal.

Block of intercellular communication: interaction of intracellular H+ and Ca2+

1987 ◽  
Vol 253 (4) ◽  
pp. C607-C612 ◽  
Author(s):  
J. M. Burt
Keyword(s):  
Intercellular Communication ◽  
Lucifer Yellow ◽  
Contractile Activity ◽  
Neonatal Rat ◽  
Dye Coupling ◽  
Myocardial Cells ◽  
Dye Transfer ◽  
Intracellular Acidosis ◽  
Communication Interaction ◽  
Junctional Permeability

The influence of elevated intracellular levels of H+ and Ca2+ on intercellular communication between cultured neonatal rat myocardial cells was examined by quantifying the percent of primary neighboring cells to which intracellularly injected Lucifer yellow had spread within 10 s of injection. Partial acidosis was induced by incubation in and then removal of NH4Cl. Intracellular Ca2+ was raised through the use of treatments that are standard in studies of heart muscle: reduction of the Na+ gradient, addition of caffeine, and combinations of these interventions. Under control conditions and during application of NH4Cl, cells exhibited spontaneous electrical and contractile activity and were well coupled (dye detectable in 100% of primary neighbors). Sustained intracellular acidosis without simultaneous elevation of intracellular Ca2+ (NH4Cl exposure followed by zero Na+, zero Ca2+) reduced the incidence of dye transfer to 90%. Elevation of intracellular Ca2+ (exposure to zero Na+, Ca2+-containing solution, with or without 10 mM caffeine) had no effect on coupling. These same interventions, when employed together, reduced the incidence of dye coupling to 18%. The results are consistent with a synergism of action of Ca2+ and H+ in the regulation of junctional permeability.

Distribution and coverage of A- and B-type horizontal cells stained with Neurobiotin in the rabbit retina

1994 ◽  
Vol 11 (3) ◽  
pp. 549-560 ◽  
Author(s):  
Stephen L. Mills ◽  
Stephen C. Massey
Keyword(s):  
Lucifer Yellow ◽  
Horizontal Cell ◽  
Horizontal Cells ◽  
Rabbit Retina ◽  
Dye Coupling ◽  
Cell Type ◽  
Axon Terminals ◽  
Visual Streak ◽  
Dendritic Fields

AbstractBoth A- and B-type horizontal cells in the rabbit retina were labeled by brief in vitro incubations of the isolated retina in the blue fluorescent dye 4,6–diamino-2–phenylindole. Intracellular injection of Lucifer Yellow into the somata revealed the morphology of the individual cells. Dye-coupling with Lucifer Yellow was seen only between A-type horizontal cells. By contrast, injection of the tracer Neurobiotin showed dye-coupling between both A- and B-type horizontal cells. There also appeared to be coupling between the axon terminals of B-type horizontal cells.The extensive dye-coupling seen following injection of Neurobiotin into a single horizontal cell soma can be used to obtain population counts of each cell type. Staining of large numbers of each cell type across the retina showed that each type increased in number and declined in dendritic diameter as the visual streak was approached, such that relatively constant coverage across the retina was maintained. In the visual streak, A-type horizontal cells numbered 555 cells/mm2 and averaged 120 μm in diameter, compared to 1375 cells/mm2 and 100 μm for B-type horizontal cells. In the periphery, the A- and B-types numbered 250 cells/mm2 and 400 cells/mm2, respectively. The average diameters of the dendritic trees at these locations were 225 μm for the A-type and 175 μm for the B-type. Coverage across the retina averaged almost six for A-type horizontal cells and 8–10 for the B-type. A-type horizontal cells in the visual streak whose elliptical dendritic fields were shown by Bloomfield (1992) to correlate physiologically with orientation bias were shown to be dye-coupled to cells with symmetrical dendritic fields.

Intracellular cyclic AMP concentration modulates gap junction permeability in parturient rat myometrium

1992 ◽  
Vol 70 (3) ◽  
pp. 358-364 ◽  
Author(s):  
Nobuyoshi Sakai ◽  
Michael G. Blennerhassett ◽  
Robert E. Garfield
Keyword(s):  
Gap Junction ◽  
Lucifer Yellow ◽  
Contractile Activity ◽  
Input Resistance ◽  
Uterine Wall ◽  
Resting Membrane Potential ◽  
Cell Coupling ◽  
Cyclic Monophosphate ◽  
Myometrial Cells ◽  
High Concentrations

We investigated whether cell-to-cell coupling between myometrial cells of parturient rats is influenced by intracellular adenosine 3′,5′-cyclic monophosphate (cAMP) concentration. To evaluate the coupling, we measured input resistance (R0) and injected Lucifer Yellow (LY) using microelectrode techniques. The intercellular spread of the dye was then observed. Longitudinal muscle strips from rat myometrium were exposed to isoproterenol, forskolin, or dibutyryl cAMP (DB-cAMP) to elevate cAMP. Isoproterenol (10−11–10−6 M) and DB-cAMP (10−5–10−3 M) hyperpolarized the resting membrane potential (Em) and increased R0 in a dose-dependent fashion. Forskolin (10−6 M) also hyperpolarized Em and increased R0. When LY was injected into a single cell, LY spread rapidly and extensively to neighboring cells in parturient control tissues, while LY transfer was completely blocked by any of the three agents at high concentrations. The increased R0 and blocked transfer of LY owing to these agents indicate that the cell-to-cell coupling was decreased both electrically and metabolically. Myometrial cells of parturient rats show increased number and size of gap junctions (GJs). The rapid and reversible decrease in coupling is interpreted to reflect the reduced permeability of GJs between the muscle cells because of an elevation of cAMP. Control of GJ permeability by this second messenger may be important for the physiological regulation of intercellular coupling and the extent of synchronizing and coordinating electrical, metabolic, and contractile activity in the uterine wall during pregnancy and parturition.Key words: adenosine 3′,5′-cyclic monophosphate, cell-to-cell coupling, gap junction permeability, rat myometrium.

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