Coexpression of Connexin45 with Connexin43 Decreases Gap Junction Size

Grikscheit, Katharina; Thomas, Neil; Bruce, Alexandra F.; Rothery, Stephen; Chan, Joachim; Severs, Nicholas J.; Dupont, Emmanuel

doi:10.1080/15419060802013943

Cited by 25 publications

(21 citation statements)

References 30 publications

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“…In previous studies, we found that altering connexin co-expression ratios in our cell lines directly changed the composition of gap junction channels at cell interfaces, and their cell-to-cell electrical conductance and dye permeability properties [8,17]. A noteworthy observation was that altered expression of the transfected connexins appeared to affect expression of the endogenous Cx43.…”

Section: Introductionmentioning

confidence: 59%

“…RLE cells, engineered with the Ecdysone-Inducible Expression System (Invitrogen, CA, USA) to co-express Cx40, ind40 cells, or Cx45, ind45 cells, with endogenous Cx43, were cultured as described previously [8,17]. The mRNA expressed upon induction is a bi-cistronic mRNA, with an internal ribosome entry site between the connexin sequence and the antibiotic resistance.…”

Section: Routine Cell Culturementioning

confidence: 99%

“…The maintenance medium contained 2 µM Ponasterone A (Pon-A) and 250 µg/ml hygromycinB to eliminate cells that lost inducibility. See [17] for a generic map of our constructs. To induce Cx40 or Cx45 expression, Pon-A was added to medium to a maximum concentration of 2 µM and incubated for ≥12 h in the absence of selection.…”

Section: Routine Cell Culturementioning

confidence: 99%

“…tagged connexin proteins and an antibody calibration technique described previously [8,17]. Immunoreactions were normalized to total protein, which relates to cellular volume.…”

Section: Protein Extraction and Western Blottingmentioning

confidence: 99%

“…RLE cells were transfected to express variable levels of either Cx40 (ind40 cells) or Cx45 (ind45 cells) under the control of an exogenous inducible non-mammalian expression system (ecdysone system) against a background of endogenous Cx43 [8,17] .…”

Section: Introductionmentioning

confidence: 99%

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Functional consequences of co-expressing connexin40 or connexin45 with connexin43 on intercellular electrical coupling

Thomas

Gray

Fry

et al. 2017

Biochemical and Biophysical Research Communications

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The functional characteristics of the co-expression of connexin43, connexin40, and connexin45 proteins in human myocardium are thought to play an important role in governing normal propagation of the cardiac electrical impulse and in generating the myocardial substrate for some arrhythmias and conduction disturbances.A rat liver epithelial cell line, that endogenously expresses connexin43, was used to induce also expression of connexin40 or connexin45 after stable transfection using an inducible ecdysone system. Electrical coupling was estimated from measurement of the input resistance of transfected cells using an intracellular microelectrode to inject current and record changes to membrane potential.However, varied expression of the transfected connexin40 or connexin45 did not change electrical coupling, although connexin43/40 co-expression led to better coupling than connexin43/45 co-expression. Quantification of endogenous connexin43 expression, at both mRNA and protein levels, showed that it was altered in a manner dependent on the transfected connexin isotype.The data using rat liver epithelial cells indicate an increased electrical coupling upon expression of connexin40 and connexin43 but decreased coupling with connexin45 and connexin43 co-expression.

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Section: Introductionmentioning

confidence: 59%

Section: Routine Cell Culturementioning

confidence: 99%

Section: Routine Cell Culturementioning

confidence: 99%

“…tagged connexin proteins and an antibody calibration technique described previously [8,17]. Immunoreactions were normalized to total protein, which relates to cellular volume.…”

Section: Protein Extraction and Western Blottingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Functional consequences of co-expressing connexin40 or connexin45 with connexin43 on intercellular electrical coupling

Thomas

Gray

Fry

et al. 2017

Biochemical and Biophysical Research Communications

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The effects of polyunsaturated fatty acids and antioxidant vitamins on atrial oxidative stress, nitrotyrosine residues, and connexins following extracorporeal circulation in patients undergoing cardiac surgery

et al. 2017

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Cardiac surgery with extracorporeal circulation is characterized by different degrees of myocardial ischemia/reperfusion, which is often associated with postoperative atrial fibrillation (POAF). We have previously shown that a novel preventive therapy based on the reinforcement of the antioxidant system using omega-3 fatty acids plus antioxidant vitamin supplementation applied to patients undergoing cardiac surgery reduces POAF occurrence. We hypothesized that oxidative stress and nitrosative stress are involved in the development of an arrhythmogenic substrate by their effect on connexins (Cx40, Cx43 and Cx45) abundance and distribution pattern. Therefore, we have assessed the effect of redox status on atrial tissue in patients undergoing cardiac surgery. Placebo/POAF and supplemented/POAF patients showed 276 and 170% higher reactive oxygen species (ROS) levels and 223 and 96% higher nitrotyrosine residues levels, respectively, compared to sinus rhythm (SR). In POAF tissue, antioxidant supplementation prevented Cx40 and Cx43 lateralization on cardiomyocyte sarcolemma, keeping them at the intercalated disks. POAF samples showed Cx40 heterogeneous distribution pattern, presenting tissue areas lacking this protein (49 and 55% lower levels in placebo/POAF and supplemented/POAF groups, respectively, compared to SR). Of note, Cx45 overexpression occurred in POAF, being 211 and 167% higher in placebo/POAF and supplemented/POAF groups, respectively, compared to SR. It is concluded that treatment with omega-3 fatty acids and antioxidant vitamins reduces oxidative and nitrosative stress and prevents Cx40/Cx43 lateralization in atrial tissue likely contributing to POAF prevention. However, it failed to fully prevent POAF occurrence because these compounds have no effects on the normalization of Cx40 down-regulation and Cx45 up-regulation, which may promote POAF.

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Cell-to-Cell Communication and Impulse Propagation

Kléber¹

2018

Cardiac Electrophysiology: From Cell to Bedside

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Coexpression of Connexin45 with Connexin43 Decreases Gap Junction Size

Cited by 25 publications

References 30 publications

Functional consequences of co-expressing connexin40 or connexin45 with connexin43 on intercellular electrical coupling

Functional consequences of co-expressing connexin40 or connexin45 with connexin43 on intercellular electrical coupling

The effects of polyunsaturated fatty acids and antioxidant vitamins on atrial oxidative stress, nitrotyrosine residues, and connexins following extracorporeal circulation in patients undergoing cardiac surgery

Cell-to-Cell Communication and Impulse Propagation

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