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.

Effect of gap junction number and permeability on intercellular coupling in rat myometrium

1991 ◽  
Vol 261 (6) ◽  
pp. C1001-C1009 ◽  
Author(s):  
M. G. Blennerhassett ◽  
R. E. Garfield
Keyword(s):  
Gap Junctions ◽  
Gap Junction ◽  
Lucifer Yellow ◽  
Input Resistance ◽  
Cell Coupling ◽  
Intercellular Coupling ◽  
Myometrial Cells ◽  
Intercellular Spread

Gap junctions (GJ) increase between myometrial cells immediately before labor. To provide evidence of their role in cell-to-cell coupling, we evaluated input resistance (Ro) and intercellular spread of Lucifer yellow (LY) in intact preparations of rat longitudinal myometrium of preterm, term, and antiprogesterone-treated preterm delivering animals. LY injected into cells from either term or preterm delivering rats (many GJ) spread rapidly to neighboring cells by 60 s, but in preparations from nondelivering controls required 4-6 min to become detectable in adjacent cells. Ro of cells in preterm nondelivering preparations was 24.1 +/- 0.8 (SE) M omega, but dropped to 12.0 +/- 0.4 M omega (P less than 0.05) at delivery, similar to preterm delivering tissues at 13.8 +/- 0.6 M omega. The putative GJ uncoupling agent octanol reversibly increased Ro of term- and preterm-delivering tissues fourfold (P less than 0.01) within 60 s, and Ro of preterm-nondelivering tissue was further increased so that Ro values were similar among the three classes. These increased Ro values are interpreted as decreased coupling. Both K+ depolarization and oxytocin (10(-8) to 10(-6) M) increased Ro of delivering tissues (P less than 0.05), suggesting that high levels of contractile agonists may lead to reduced cell-to-cell coupling. Therefore, myometrial coupling can be modulated over seconds via GJ permeability as well as over hours by GJ number.

scholarly journals Descending vasa recta endothelial cells and pericytes form mural syncytia

2014 ◽  
Vol 306 (7) ◽  
pp. F751-F763 ◽  
Author(s):  
Zhong Zhang ◽  
Hai Lin ◽  
Chunhua Cao ◽  
Kristie Payne ◽  
Thomas L. Pallone
Keyword(s):  
Endothelial Cells ◽  
Membrane Potential ◽  
Patch Clamp ◽  
Gap Junction ◽  
Lucifer Yellow ◽  
Sampling Rate ◽  
Input Resistance ◽  
Contralateral Side ◽  
Vasa Recta ◽  
Endothelial Response

Using patch clamp, we induced depolarization of descending vasa recta (DVR) pericytes or endothelia and tested whether it was conducted to distant cells. Membrane potential was measured with the fluorescent voltage dye di-8-ANEPPS or with a second patch-clamp electrode. Depolarization of an endothelial cell induced responses in other endothelia within a millisecond and was slowed by gap junction blockade with heptanol. Endothelial response to pericyte depolarization was poor, implying high-resistance myo-endothelial coupling. In contrast, dual patch clamp of neighboring pericytes revealed syncytial coupling. At high sampling rate, the spread of depolarization between pericytes and endothelia occurred in 9 ± 2 or 12 ± 2 μs, respectively. Heptanol (2 mM) increased the overall input resistance of the pericyte layer to current flow and prevented transmission of depolarization between neighboring cells. The fluorescent tracer Lucifer yellow (LY), when introduced through ruptured patches, spread between neighboring endothelia in 1 to 7 s, depending on location of the flanking cell. LY diffused to endothelial cells on the ipsilateral but not contralateral side of the DVR wall and minimally between pericytes. We conclude that both DVR pericytes and endothelia are part of individual syncytia. The rate of conduction of membrane potential exceeds that for diffusion of hydrophilic molecules by orders of magnitude. Gap junction coupling of adjacent endothelial cells may be spatially oriented to favor longitudinal transmission along the DVR axis.

Wortmannin inhibits contraction without altering electrical activity in canine gastric smooth muscle

1996 ◽  
Vol 270 (5) ◽  
pp. C1405-C1412 ◽  
Author(s):  
E. P. Burke ◽  
W. T. Gerthoffer ◽  
K. M. Sanders ◽  
N. G. Publicover
Keyword(s):  
Membrane Potential ◽  
Light Chain ◽  
Myosin Light Chain ◽  
Contractile Activity ◽  
Smooth Muscles ◽  
Slow Waves ◽  
Resting Membrane Potential ◽  
Partial Recovery ◽  
Gastric Smooth Muscle ◽  
High Concentrations

Wortmannin, an inhibitor of myosin light-chain kinase (10-30 microM), completely and irreversibly abolished (in 75% of tissues from canine gastric antrum) phase contractions caused by slow waves with no significant effects on resting membrane potential or the frequency, amplitude, or duration of spontaneous slow waves. Responses to agents that normally cause hyperpolarization (cromakalim, sodium nitroprusside, and forskolin) were unaffected by wortmannin treatment. It was also possible to study the excitatory effects of agents and conditions that normally result in loss of intracellular impalements: 1) elevated extracellular K+ concentrations altered membrane potential close to values predicted by the Nernst equation, and 2) high concentrations of acetylcholine produced depolarization and rapid oscillations in membrane potential coincident with contractile activity. Cholinergic increases in myosin light-chain phosphorylation and contractions were partially blocked by wortmannin. In canine antrum, wortmannin inhibition of contraction was irreversible, although in other tissue types, partial recovery of contractions was observed when wortmannin was removed. Wortmannin can be a useful agent to investigate the electrophysiology of some smooth muscles when movement might lead to recording artifacts or loss of signal.

Characterization of gap junction channels in A7r5 vascular smooth muscle cells

1991 ◽  
Vol 260 (5) ◽  
pp. C975-C981 ◽  
Author(s):  
L. K. Moore ◽  
E. C. Beyer ◽  
J. M. Burt
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Gap Junctions ◽  
Gap Junction ◽  
Second Messengers ◽  
Protein Composition ◽  
Cell Coupling ◽  
Cyclic Monophosphate ◽  
A7r5 Cells ◽  
Immunohistochemical Studies

Recent evidence suggest that coordination of blood flow in the microcirculation involves cell-to-cell coupling via gap junctions. In this study, using A7r5 cells as a model of vascular smooth muscle, we have characterized the gap junctions in terms of the unitary conductances of the observed channels, the responses to second messengers, and subunit protein composition. The cells were typically well coupled several hours after plating, with junctional conductances on the order 20-40 nS. Channels with mean conductances of 36 and 89 pS were observed in low-conductance cell pairs and in cell pairs whose macroscopic conductance was reduced by exposure to halothane. Connexin43 was the only known gap junction sequence detected by Northern blots (low and high stringency), immunoblots, or immunohistochemical studies. Junctional conductance was reduced 15% by 8-bromoadenosine 3',5'-cyclic monophosphate; 8-bromoguanosine 3',5'-cyclic monophosphate had no effect. The results suggest that connexin43 can form stable channels of at least two distinct conductances and gap junctions with differing responses to second messengers.

Structure-activity relations of the cardiac gap junction channel

1990 ◽  
Vol 258 (2) ◽  
pp. C195-C205 ◽  
Author(s):  
D. C. Spray ◽  
J. M. Burt
Keyword(s):  
Gap Junction ◽  
Cyclic Nucleotides ◽  
Rat Heart ◽  
Unsaturated Fatty Acids ◽  
Lucifer Yellow ◽  
Cyclooxygenase Inhibitors ◽  
Cyclic Monophosphate ◽  
Gap Junction Channel ◽  
Junction Protein ◽  
Sites Of Action

Cardiac gap junction channels play the important roles of synchronizing pacemaker cells and allowing impulse propagation along the conduction system and throughout the ventricular myocardium. These channels, which support current flow in both longitudinal and tranverse directions, are permeable to anions and cations with radii less than approximately 0.5 nm and in rat heart have unitary conductances on the order of 50 pS. This unitary conductance is consistent with channel geometry described by a right cylindrical pore with diameter large enough for the brilliantly fluorescent dye molecule lucifer yellow to pass between cells. These channels, like others in biological systems, are opened and closed by various treatments, a process termed gating. Cytoplasmic acidification reduces junctional conductance (gj), an effect that is apparently potentiated by elevated myoplasmic Ca ions. Reduced gj also occurs in response to a variety of lipophilic molecules, including halothane, heptanol, and unsaturated fatty acids; the mechanism of action may involve disruption of the protein-lipid microenvironment of the gap junction channel. Arachidonic acid uncouples, and this effect is partially, but incompletely, blocked by an inhibitor of the lipoxygenase metabolic pathways. Cyclooxygenase inhibitors have no protective effects. Certain cyclic nucleotides can rapidly increase gj [adenosine 3',5'-cyclic monophosphate (cAMP)] or slightly decrease it [guanosine 3',5'-cyclic monophosphate (cGMP)], and agents that use these cyclic nucleotides as second messengers (isoproterenol and perhaps carbachol, respectively) produce consistent effects. Agents expected to cause protein kinase C activation (tumor-promoting phorbol esters and diacylglycerol) increase gj rapidly. The gap junction protein from rat heart has been cloned and sequenced. From the primary sequence for the protein, plausible sites of action within the putative cytoplasmic domains are proposed for each of these treatments. In response to gating stimuli that close the channel (halothane, CO2, heptanol), unitary channel conductance is unchanged, suggesting that these agents act by reducing open time probability. Together, these properties constitute the beginnings of our endeavor to define pharmacological agents that are potentially useful in therapeutic manipulation of synchronous discharge, conduction velocity, and isochronous wavefront propagation in cardiac tissue.

Dorsal and Ventral Distribution of Excitable and Synaptic Properties of Neurons of the Bed Nucleus of the Stria Terminalis

2003 ◽  
Vol 90 (1) ◽  
pp. 405-414 ◽  
Author(s):  
Regula E. Egli ◽  
Danny G. Winder
Keyword(s):  
Receptor Antagonist ◽  
Drugs Of Abuse ◽  
Input Resistance ◽  
Membrane Properties ◽  
Resting Membrane Potential ◽  
Stria Terminalis ◽  
Bed Nucleus ◽  
Adult Mice ◽  
Dependent Potential

The bed nucleus of the stria terminalis (BNST) is a structure uniquely positioned to integrate stress information and regulate both stress and reward systems. Consistent with this arrangement, evidence suggests that the BNST, and in particular the noradrenergic input to this structure, is a key component of affective responses to drugs of abuse. We have utilized an in vitro slice preparation from adult mice to determine synaptic and membrane properties of these cells, focusing on the dorsal and ventral subdivisions of the anterolateral BNST (dBNST and vBNST) because of the differential noradrenergic input to these two regions. We find that while resting membrane potential and input resistance are comparable between these subdivisions, excitable properties, including a low-threshold spike (LTS) likely mediated by T-type calcium channels and an Ih-dependent potential, are differentially distributed. Inhibitory and excitatory postsynaptic potentials (IPSPs and EPSPs, respectively) are readily evoked in both dBNST and vBNST. The fast IPSP is predominantly GABAA-receptor mediated and is partially blocked by the AMPA/kainate-receptor antagonist CNQX. In the presence of the GABAA-receptor antagonist picrotoxin, cells in dBNST but not vBNST are more depolarized and have a higher input resistance, suggesting tonic GABAergic inhibition of these cells. The EPSPs elicited in BNST are monosynaptic, exhibit paired pulse facilitation, and contain both an AMPA- and an N-methyl-d-aspartate (NMDA) receptor-mediated component. These data support the hypothesis that neurons of the dorsal and ventral BNST differentially integrate synaptic input, which is likely of behavioral significance. The data also suggest mechanisms by which information may flow through stress and reward circuits.

Postnatal Changes in Membrane Properties of Mice Trigeminal Ganglion Neurons

2002 ◽  
Vol 87 (5) ◽  
pp. 2398-2407 ◽  
Author(s):  
Carmen Cabanes ◽  
Mikel López de Armentia ◽  
Félix Viana ◽  
Carlos Belmonte
Keyword(s):  
Postnatal Development ◽  
Trigeminal Ganglion ◽  
Input Resistance ◽  
Membrane Capacitance ◽  
Membrane Properties ◽  
Inward Rectification ◽  
Resting Membrane Potential ◽  
Depolarization Rate ◽  
Ganglion Neurons

Intracellular recordings from neurons in the mouse trigeminal ganglion (TG) in vitro were used to characterize changes in membrane properties that take place from early postnatal stages (P0–P7) to adulthood (>P21). All neonatal TG neurons had uniformly slow conduction velocities, whereas adult neurons could be separated according to their conduction velocity into Aδ and C neurons. Based on the presence or absence of a marked inflection or hump in the repolarization phase of the action potential (AP), neonatal neurons were divided into S- (slow) and F-type (fast) neurons. Their passive and subthreshold properties (resting membrane potential, input resistance, membrane capacitance, and inward rectification) were nearly identical, but they showed marked differences in AP amplitude, AP overshoot, AP duration, rate of AP depolarization, rate of AP repolarization, and afterhyperpolarization (AHP) duration. Adult TG neurons also segregated into S- and F-type groups. Differences in their mean AP amplitude, AP overshoot, AP duration, rate of AP depolarization, rate of AP repolarization, and AHP duration were also prominent. In addition, axons of 90% of F-type neurons and 60% of S-type neurons became faster conducting in their central and peripheral branch, suggestive of axonal myelination. The proportion of S- and F-type neurons did not vary during postnatal development, suggesting that these phenotypes were established early in development. Membrane properties of both types of TG neurons evolved differently during postnatal development. The nature of many of these changes was linked to the process of myelination. Thus myelination was accompanied by a decrease in AP duration, input resistance ( R in), and increase in membrane capacitance (C). These properties remained constant in unmyelinated neurons (both F- and S-type). In adult TG, all F-type neurons with inward rectification were also fast-conducting Aδ, suggesting that those F-type neurons showing inward rectification at birth will evolve to F-type Aδ neurons with age. The percentage of F-type neurons showing inward rectification also increased with age. Both F- and S-type neurons displayed changes in the sensitivity of the AP to reductions in extracellular Ca2+ or substitution with Co2+ during the process of maturation.

scholarly journals Gap junction structures after experimental alteration of junctional channel conductance.

1985 ◽  
Vol 101 (5) ◽  
pp. 1741-1748 ◽  
Author(s):  
T M Miller ◽  
D A Goodenough
Keyword(s):  
Gap Junctions ◽  
Gap Junction ◽  
Electron Micrographs ◽  
Time Course ◽  
Structural Changes ◽  
Lucifer Yellow ◽  
Freeze Fracture ◽  
Channel Conductance ◽  
Experimental Alteration ◽  
Quick Freezing

Gap junctions are known to present a variety of different morphologies in electron micrographs and x-ray diffraction patterns. This variation in structure is not only seen between gap junctions in different tissues and organisms, but also within a given tissue. In an attempt to understand the physiological meaning of some aspects of this variability, gap junction structure was studied following experimental manipulation of junctional channel conductance. Both physiological and morphological experiments were performed on gap junctions joining stage 20-23 chick embryo lens epithelial cells. Channel conductance was experimentally altered by using five different experimental manipulations, and assayed for conductance changes by observing the intercellular diffusion of Lucifer Yellow CH. All structural measurements were made on electron micrographs of freeze-fracture replicas after quick-freezing of specimens from the living state; for comparison, aldehyde-fixed specimens were measured as well. Analysis of the data generated as a result of this study revealed no common statistically significant changes in the intrajunctional packing of connexons in the membrane plane as a result of experimental alteration of junctional channel conductance, although some of the experimental manipulations used to alter junctional conductance did produce significant structural changes. Aldehyde fixation caused a dramatic condensation of connexon packing, a result not observed with any of the five experimental uncoupling conditions over the 40-min time course of the experiments.

Effect of aminophylline on the contraction threshold of rat diaphragm fibers

1991 ◽  
Vol 71 (4) ◽  
pp. 1409-1414 ◽  
Author(s):  
A. S. Losavio ◽  
B. A. Kotsias
Keyword(s):  
Membrane Potential ◽  
Sarcoplasmic Reticulum ◽  
Resting Membrane Potential ◽  
Dependent Manner ◽  
Direct Visualization ◽  
Rat Diaphragm ◽  
Current Clamp ◽  
Cyclic Monophosphate ◽  
Dose Dependent

We studied the effect of aminophylline (0.1–1 mM) on the contraction threshold (CT) of rat diaphragm fibers (25 degrees C). The CT was measured by direct visualization (x200) of the fiber under current-clamp conditions. The main findings are the following: 1) Aminophylline lowers the CT, in a dose-dependent manner, toward more negative values of the resting membrane potential (Vm). 2) Dibutyryl adenosine 3′,5′-cyclic monophosphate (2 mM) shifts the CT, although this change is smaller than in the presence of xanthine. 3) Tetracaine (1 mM), a drug that diminishes Ca release from the sarcoplasmic reticulum, reduces the shift induced by 1 mM aminophylline; this is partially overcome by increasing aminophylline concentration to 5 mM. 4) Hyperpolarization of the fibers shifts the CT to more negative Vm. We suggest that the displacement in the CT to more negative Vm plays an important role in the potentiating effect of aminophylline. This could be the result of an enhancement of Ca release from the sarcoplasmic reticulum.

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