Effects of airway surface liquid height on the kinetics of extracellular nucleotides in airway epithelia

Amarante, Tauanne Dias; Silva, J. Kamphorst Leal da; Garcia, Guilherme J. M.

doi:10.1016/j.jtbi.2014.08.030

Cited by 3 publications

(5 citation statements)

References 18 publications

(28 reference statements)

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“…Airway surface liquid (ASL) was visualized by adding 10 kD-Texas Reddextran (ThermoFisher) to the apical surface of NHBE or HBEC3-KT cell cultures at ALI, as previously described (Amarante et al, 2014;Button et al, 2008;Lazarowski et al, 2004), and z-stack ASL images were collected 4 h later with a confocal microscope at 37°C, 5% CO 2 . The apical surface of cells was washed once with PBS 18 h prior to the addition of Texas Red.…”

Section: Measuring Airway Surface Liquid Heightmentioning

confidence: 99%

“…The apical surface of cells was washed once with PBS 18 h prior to the addition of Texas Red. ASL was visualized by adding 10 kD Texas Red-dextran (17.5 µl per 12-well insert and 6.7 µl per 24-well insert at 1.5 mg/ml in sterile PBS, ThermoFisher) to the apical surface of NHBE or HBEC3-KT cell cultures at ALI, as previously described (Amarante et al, 2014;Button et al, 2008;Lazarowski et al, 2004). Fluorinert (100 µl per 12-well insert and 30 µl per 24-well insert) was added to the top of the insert cultures to prevent evaporation.…”

Section: Measuring Airway Surface Liquid Heightmentioning

confidence: 99%

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Adenine nucleotide translocase regulates airway epithelial metabolism, surface hydration and ciliary function

Kliment

Nguyen

Kaltreider

et al. 2021

Journal of Cell Science

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Airway hydration and ciliary function are critical to airway homeostasis and dysregulated in chronic obstructive lung disease. COPD is impacted by cigarette smoking with no therapeutic options. We utilized a high copy cDNA library genetic selection approach in the amoeba Dictyostelium discoideum to identify genetic protectors from cigarette smoke (CS). Adenine nucleotide translocase (Salvioli et al.), a mitochondrial ADP/ATP transporter, was protective against CS in Dictyostelium and human bronchial epithelial cells. ANT2 gene expression is reduced in lung tissue from COPD patients and in a mouse smoking model. ANT1 and ANT2 overexpression resulted in enhanced oxidative respiration and ATP flux. In addition to ANT's presence in the mitochondria, ANT resides at the plasma membrane in airway epithelial cells and regulates airway homeostasis. ANT2 overexpression stimulates airway surface hydration by ATP and maintains ciliary beating after CS exposure, which are key functions of the airway. Our study highlights the potential of ANT upregulation and/or agonists in protecting from dysfunctional mitochondrial metabolism, airway hydration, and ciliary motility in COPD.

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Section: Measuring Airway Surface Liquid Heightmentioning

confidence: 99%

Section: Measuring Airway Surface Liquid Heightmentioning

confidence: 99%

Adenine nucleotide translocase regulates airway epithelial metabolism, surface hydration and ciliary function

Kliment

Nguyen

Kaltreider

et al. 2021

Journal of Cell Science

View full text Add to dashboard Cite

show abstract

“…ASL was visualized by adding 10 kD-Texas Red-dextran (ThermoFisher) to the apical surface of NHBE or HBEKT cultures at air liquid interface, as previously described [23,25,42] and z-stack ASL images collected 4 hr later on confocal microscope at 37°C, 5% CO2. The apical surface of cells was washed once with PBS 18 hr prior to the addition of Texas Red.…”

Section: Measuring Airway Surface Liquid (Asl) Heightmentioning

confidence: 99%

“…The apical surface of cells was washed once with PBS 18 hr prior to the addition of Texas Red. ASL was visualized by adding 10 kD Texas Red-dextran (17.5 µl per 12-well insert and 6.7 µl per 24-well insert at 1.5 mg/mL in sterile PBS, ThermoFisher) to the apical surface of NHBE or HBEKT cultures at air liquid interface, as previously described [23,25,42]. Fluorinert (100 µl per 12-well insert and 30 µl per 24-well insert) was added to the top of the insert cultures to prevent evaporation.…”

Section: Measuring Airway Surface Liquid (Asl) Heightmentioning

confidence: 99%

Adenine Nucleotide Translocase regulates the airway epithelium, mitochondrial metabolism and ciliary function

Kliment

Nguyen

Kaltreider

et al. 2020

Preprint

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The authors have declared no conflicts of interest exist. AbstractAirway hydration and ciliary function are critical to airway homeostasis and dysregulated in chronic obstructive lung disease (COPD). COPD is the 4 th leading cause of death in the US and is impacted by cigarette smoking with no therapeutic options. We utilized a genetic selection approach in the amoeba Dictyostelium discoideum as a comparative discovery tool in lung biology to identify genetic protectors from cigarette smoke (CS). Adenine nucleotide translocase (ANT), a mitochondrial ADP/ATP transporter, was protective against CS in Dictyostelium and human bronchial epithelial cells. ANT2 gene expression is reduced in lung tissue from COPD patients and in a mouse smoking model. ANT1and ANT2 overexpression resulted in enhanced oxidative respiration and ATP flux. In addition to ANT's presence in the mitochondria, ANT1 and ANT2 reside at the plasma membrane in airway epithelial cells and this localization plays a role in how ANTs regulate airway homeostasis. ANT2 overexpression stimulates airway surface liquid hydration by ATP and maintains ciliary beating after CS exposure, which are key functions of the airway. Our study highlights the potential of ANT modulation in protecting from dysfunctional mitochondrial metabolism, airway hydration, and ciliary motility in COPD.

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“…For comparison, the levels of adenosine in human plasma are typically measured at less than 200 nM, but can be in excess of 500 nM in people with sickle cell disease. (Ramakers et al 2008; Zhang et al 2011) Airway surface liquid is normally 150–200 nM adenosine (Zuo et al 2008) although recent evidence suggests that at the cell surface, where many of the regulatory enzymes and receptors are positioned, the concentration may be 5 to 10 times higher (Amarante et al 2014). …”

Section: Introductionmentioning

confidence: 99%

A metabolomic, geographic, and seasonal analysis of the contribution of pollen-derived adenosine to allergic sensitization

et al. 2016

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Background Studies on ragweed and birch pollen extracts suggested that the adenosine content is an important factor in allergic sensitization. However, exposure levels from other pollens and considerations of geographic and seasonal factors have not been evaluated. Objective This study compared the metabolite profile of pollen species important for allergic disease, specifically measured the adenosine content, and evaluated exposure to pollen-derived adenosine. Methods An NMR metabolomics approach was used to measure metabolite concentrations in twenty-six pollen extracts. Pollen count data was analyzed from five cities to model exposure. Results A principal component analysis of the various metabolites identified by NMR showed that pollen extracts could be differentiated primarily by sugar content: glucose, fructose, sucrose, and myo-inositol. In extracts of 10 mg of pollen/ml, the adenosine was highest for grasses (45 μM) followed by trees (23 μM) and weeds (19 μM). Pollen count data showed that tree pollen was typically 5–10 times the amount of other pollens. At the daily peaks of tree, grass, and weed season the pollen-derived adenosine exposure per day is likely to only be 1.1, 0.11, and 0.12 μg, respectively. Seasonal models of pollen exposure and respiration suggest that it would be a rare event limited to tree pollen season for concentrations of pollen-derived adenosine to approach physiological levels. Conclusions Sugar content and other metabolites may be useful in classifying pollens. Unless other factors create localized exposures that are very different from these models, pollen-derived adenosine is unlikely to be a major factor in allergic sensitization.

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Effects of airway surface liquid height on the kinetics of extracellular nucleotides in airway epithelia

Cited by 3 publications

References 18 publications

Adenine nucleotide translocase regulates airway epithelial metabolism, surface hydration and ciliary function

Adenine nucleotide translocase regulates airway epithelial metabolism, surface hydration and ciliary function

Adenine Nucleotide Translocase regulates the airway epithelium, mitochondrial metabolism and ciliary function

A metabolomic, geographic, and seasonal analysis of the contribution of pollen-derived adenosine to allergic sensitization

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