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Results: 7

1.
Fig. 7.

Fig. 7. From: Enhancement of alveolar epithelial sodium channel activity with decreased cystic fibrosis transmembrane conductance regulator expression in mouse lung.

Cftr mRNA is expressed in ATII cells. Shown are the products from RT-PCR analysis by using whole wild-type mouse lung or highly purified ATII cells. Lanes 1-4 represent reactions using mouse Cftr primers. Lanes 1 and 2 are samples from wild-type+/+ and Cftr−/− mouse lungs, respectively. Lane 3 is a sample from highly purified ATII cells isolated from Cftr+/+ mouse lungs. Lane 4 represents a nontemplate PCR control (ntc). Lanes 5-8 represent reactions using mouse Gapdh primers from the same samples described in lanes 1-4. The Cftr PCR product is 158 bp and the Gapdh PCR product is 168 bp.

Ahmed Lazrak, et al. Am J Physiol Lung Cell Mol Physiol. 2011 October;301(4):L557-L567.
2.
Fig. 3.

Fig. 3. From: Enhancement of alveolar epithelial sodium channel activity with decreased cystic fibrosis transmembrane conductance regulator expression in mouse lung.

ENaC activity in ATII cells from ΔF508 and Cftr−/− mice. Lung slices were prepared from the lungs of as detailed in materials and methods; ATII cells were patched in the cell-attached mode in situ. A and C: recordings showing ENaC activity in ATII cells from ΔF508 and Cftr−/− mice, respectively, for ∼5 min. The membrane patch under the pipette was held at −100 mV and the lung slice was bathed in a high-K+ solution. B and D: corresponding all-events distribution histograms. The histogram in B (ΔF508) shows the presence of 1 channel with an amplitude of 0.48 pA, Po=0.6, and g=4.8 pS. The corresponding values for the histogram in D (Cftr−/−) were amplitude 0.54 pA; Po=0.86, and g=5.4 pS. Similar recordings were obtained from 5 different mice from each group, with 3–5 recordings per mouse.

Ahmed Lazrak, et al. Am J Physiol Lung Cell Mol Physiol. 2011 October;301(4):L557-L567.
3.
Fig. 1.

Fig. 1. From: Enhancement of alveolar epithelial sodium channel activity with decreased cystic fibrosis transmembrane conductance regulator expression in mouse lung.

Alveolar type I (ATI) and type II (ATII) cells in wild-type mouse lungs express 2 Na+ conductances. A and C: typical records from cell attached patches in ATII and ATI cells, respectively, showing the presence of 2 distinct amplitudes (the first at 0.44 pA and the second at 1.8 pA). The corresponding conductances were 4. 4 and 18 pS. Recordings were obtained at a membrane potential of −100 mV. B and D: corresponding event distribution histograms of the 2 amplitudes. The open probabilities (Po) of the 2 conductances are shown above their corresponding peaks. These are typical records of data obtained from at least 20 patches from 5 different wild-type mice.

Ahmed Lazrak, et al. Am J Physiol Lung Cell Mol Physiol. 2011 October;301(4):L557-L567.
4.
Fig. 4.

Fig. 4. From: Enhancement of alveolar epithelial sodium channel activity with decreased cystic fibrosis transmembrane conductance regulator expression in mouse lung.

18-pS channel activity in ATII cells from Cftr+/+ and Cftr−/− mice. Lung slices were prepared from the lungs of as detailed in materials and methods; ATII cells were patched in the cell-attached mode in situ. A and B: 18-pS channel cation activity in ATII cells from Cftr+/+ and Cftr−/− mice, respectively, for ∼5 min. The membrane patches under the pipette were held at −100 mV and the lung slices were bathed in a high-K+ solution. C and D: corresponding all-events distribution histograms from Cftr+/+ and Cftr−/− mice, respectively. The histogram in C shows the presence of 1 channel with an amplitude of 1.8 pA, Po=0.27, and g=17.7 pS. The corresponding values for the histogram in D (Cftr−/−) are amplitude 1.88 pA; Po=0.58, and g=18.8 pS. Similar recordings were obtained from 5 different mice from each group, with 3–5 recordings per mouse. E: mean values of Po (± SE) for Cftr+/+ and Cftr−/− mice; n=4. Values are significantly different from each other (P < 0.05).

Ahmed Lazrak, et al. Am J Physiol Lung Cell Mol Physiol. 2011 October;301(4):L557-L567.
5.
Fig. 2.

Fig. 2. From: Enhancement of alveolar epithelial sodium channel activity with decreased cystic fibrosis transmembrane conductance regulator expression in mouse lung.

Amiloride-sensitive sodium channel (ENaC) activity in ATII cells from Cftr+/+ and Cftr−/+ mice. Lung slices were prepared from the lungs of wild-type and Cftr−/+ mice as detailed in materials and methods; ATII cells were patched in cell-attached mode in situ. A and C: recordings from Cftr+/+ and Cftr−/+ ATII cells, respectively, showing channel openings (downward deflections) for ∼5 min when the membrane patch was held at −100 mV and the cell was depolarized to 0 mV by using high K+ in the bathing solution. B and D: corresponding event distribution histograms. The histogram in B (wild-type) shows the activity of 1 channel with amplitude of 0.49 pA, Po of 0.26, and conductance (g) of 4.9 pS. The corresponding values for the histogram in D (Cftr−/+) were amplitude 0.435; Po=0.50; g=4.35 pS. Similar recordings were obtained from at least 9 patches from 5 of wild-type and 5 Cftr−/+ mice.

Ahmed Lazrak, et al. Am J Physiol Lung Cell Mol Physiol. 2011 October;301(4):L557-L567.
6.
Fig. 6.

Fig. 6. From: Enhancement of alveolar epithelial sodium channel activity with decreased cystic fibrosis transmembrane conductance regulator expression in mouse lung.

Lung α-ENaC levels. Equal amounts of lung proteins (150–200 μg) were loaded in 10% Tris·HCl Criterion precast gels, transferred to polyvinylidene difluoride membranes, and immunostained with α-ENaC antibodies (Thermo Scientific, Rockford, IL). Bands were detected by the LumiGLO Western blot kit. A: 1=wild-type; 2=Cftr−/+; 3=Cftr−/−; 4=ΔF508. Representative Western blot was repeated 4 times in lungs from different mice. B: membranes were striped and blotted with equal concentration of an antibody against α-ENaC in the presence of blocking peptide (BP). C: mean values (± 1 SE; n=4) of band density for the indicated molecular masses (in kDa), normalized to α-tubulin. wt, Wild type. D: mean values (1 SE; n=4) of total lung α-ENaC (sum of all bands) normalized to α-tubulin. *P < 0.05 compared with the wild-type value in the same group (2-tailed P test); #P < 0.05 compared with the wild-type value in the same group (1-tailed P test).

Ahmed Lazrak, et al. Am J Physiol Lung Cell Mol Physiol. 2011 October;301(4):L557-L567.
7.
Fig. 5.

Fig. 5. From: Enhancement of alveolar epithelial sodium channel activity with decreased cystic fibrosis transmembrane conductance regulator expression in mouse lung.

Activation of ENaC in ATII cells from Cftr+/+ ΔF508 but not Cftr−/− mice by forskolin. Lung slices were prepared from each group of mice as detailed in materials and methods; ATII cells were patched in the cell-attached mode in situ; pipettes contained the adenylate cyclase activator forskolin (5 μM). AC: recordings from Cftr+/+, ΔF508, and Cftr−/− mice showing channel openings (downward deflections) for ∼5 min when the cell membrane patch was held at −100 mV and the cell was depolarized to 0 mV by incubating it in a high-K+ solution. Notice gradual increase of the current amplitude in A (Cftr+/+) as forskolin diffuses from the pipette in the cell cytoplasm. In contrast, ENaC was more constitutively active in the lungs of Cftr−/− mice and was not activated by forskolin. Solid lines slow the closed state. DF: corresponding event distribution histograms of 9 open channels for Cftr+/+, 4 for ΔF508 and 1 for Cftr−/−.

Ahmed Lazrak, et al. Am J Physiol Lung Cell Mol Physiol. 2011 October;301(4):L557-L567.

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