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Hypoxia augments the calcium-activated chloride current carried by anoctamin-1 in cardiac vascular endothelial cells of neonatal mice.

Wu MM, Lou J, Song BL, Gong YF, Li YC, Yu CJ, Wang QS, Ma TX, Ma K, Hartzell HC, Duan DD, Zhao D, Zhang ZR - Br. J. Pharmacol. (2014)

Bottom Line: The density of ICl ( C a) detected in CVECs was significantly inhibited by T16Ainh -A01, an Ano1 inhibitor, and a pore-targeting, specific anti-Ano1 antibody, and was markedly decreased in Ano1 gene knockdown CVECs.Ano1 formed CaCC in CVECs of neonatal mice.Ano1 may play a role in the pathophysiological processes during ischaemia in heart, and therefore, Ano1 might be a potential therapeutic target to prevent ischaemic damage.

View Article: PubMed Central - PubMed

Affiliation: Departments of Clinical Pharmacy and Cardiology, Institute of Clinical Pharmacy, The 2nd Affiliated Hospital, Key Laboratories of Education Ministry for Myocardial Ischemia Mechanism and Treatment, Harbin Medical University, Harbin, China.

No MeSH data available.


Related in: MedlinePlus

(A) qRT-PCR demonstrates the relative mRNA levels of Ano1 and Ano2 in CVECs (n = 6); Ano1, but not Ano2, was expressed in CVECs. (B) Western blots shown at the two left panels indicate CVECs express Ano1; with the secondary antibody alone, no band was detected (two panels at right) (repeated five times). (C) Representative immunofluorescence images of the sliced left ventricular wall in longitudinal section, labelled with antibodies raised against vWF (red) and against Ano1 (green). The white arrow heads indicate blood vessels. (D) Representative immunofluorescence images of isolated CVECs labelled with antibodies raised against CD31 (red) and against Ano1 (green). Scale bar represents 20 μm. (E) The representative fluorescence intensity histograms, constructed across a CVEC [the red solid line shown in (D)]. The majority of Ano1 (green) was distributed at and/or in the vicinity of the plasma membrane.
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fig03: (A) qRT-PCR demonstrates the relative mRNA levels of Ano1 and Ano2 in CVECs (n = 6); Ano1, but not Ano2, was expressed in CVECs. (B) Western blots shown at the two left panels indicate CVECs express Ano1; with the secondary antibody alone, no band was detected (two panels at right) (repeated five times). (C) Representative immunofluorescence images of the sliced left ventricular wall in longitudinal section, labelled with antibodies raised against vWF (red) and against Ano1 (green). The white arrow heads indicate blood vessels. (D) Representative immunofluorescence images of isolated CVECs labelled with antibodies raised against CD31 (red) and against Ano1 (green). Scale bar represents 20 μm. (E) The representative fluorescence intensity histograms, constructed across a CVEC [the red solid line shown in (D)]. The majority of Ano1 (green) was distributed at and/or in the vicinity of the plasma membrane.

Mentions: The biophysical features and pharmacological profile of the ICl(Ca) detected in CVECs are similar to those of CaCCs. We reasoned that ICl(Ca) might be mediated by Ano1. As seen in Fig. 3A, Ano1 mRNA, but not Ano2 mRNA, was detected in CVECs (Fig. 3A). Western blot analysis demonstrated that CVECs expressed Ano1 (Fig. 3B). We further stained the neonatal mouse heart with anti-Ano1 antibody and anti-vWF antibody, a marker for endothelial cells, and sliced the left ventricular walls in longitudinal sections. Ano1 (green) was nicely present at endothelium and co-localized with endothelial marker (red) (Fig. 3C; the white arrow heads indicate blood vessels). Confocal imaging analyses also revealed that a co-localization of Ano1 (green) and CD31 (red) occurred at the plasma membrane of CVECs (Fig. 3D) because the highest merged fluorescence intensity (peaks) appeared in the vicinity of the plasma membrane (Fig. 3E). These results led us to hypothesize that the ICl(Ca) is carried by Ano1 in CVECs.


Hypoxia augments the calcium-activated chloride current carried by anoctamin-1 in cardiac vascular endothelial cells of neonatal mice.

Wu MM, Lou J, Song BL, Gong YF, Li YC, Yu CJ, Wang QS, Ma TX, Ma K, Hartzell HC, Duan DD, Zhao D, Zhang ZR - Br. J. Pharmacol. (2014)

(A) qRT-PCR demonstrates the relative mRNA levels of Ano1 and Ano2 in CVECs (n = 6); Ano1, but not Ano2, was expressed in CVECs. (B) Western blots shown at the two left panels indicate CVECs express Ano1; with the secondary antibody alone, no band was detected (two panels at right) (repeated five times). (C) Representative immunofluorescence images of the sliced left ventricular wall in longitudinal section, labelled with antibodies raised against vWF (red) and against Ano1 (green). The white arrow heads indicate blood vessels. (D) Representative immunofluorescence images of isolated CVECs labelled with antibodies raised against CD31 (red) and against Ano1 (green). Scale bar represents 20 μm. (E) The representative fluorescence intensity histograms, constructed across a CVEC [the red solid line shown in (D)]. The majority of Ano1 (green) was distributed at and/or in the vicinity of the plasma membrane.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4128065&req=5

fig03: (A) qRT-PCR demonstrates the relative mRNA levels of Ano1 and Ano2 in CVECs (n = 6); Ano1, but not Ano2, was expressed in CVECs. (B) Western blots shown at the two left panels indicate CVECs express Ano1; with the secondary antibody alone, no band was detected (two panels at right) (repeated five times). (C) Representative immunofluorescence images of the sliced left ventricular wall in longitudinal section, labelled with antibodies raised against vWF (red) and against Ano1 (green). The white arrow heads indicate blood vessels. (D) Representative immunofluorescence images of isolated CVECs labelled with antibodies raised against CD31 (red) and against Ano1 (green). Scale bar represents 20 μm. (E) The representative fluorescence intensity histograms, constructed across a CVEC [the red solid line shown in (D)]. The majority of Ano1 (green) was distributed at and/or in the vicinity of the plasma membrane.
Mentions: The biophysical features and pharmacological profile of the ICl(Ca) detected in CVECs are similar to those of CaCCs. We reasoned that ICl(Ca) might be mediated by Ano1. As seen in Fig. 3A, Ano1 mRNA, but not Ano2 mRNA, was detected in CVECs (Fig. 3A). Western blot analysis demonstrated that CVECs expressed Ano1 (Fig. 3B). We further stained the neonatal mouse heart with anti-Ano1 antibody and anti-vWF antibody, a marker for endothelial cells, and sliced the left ventricular walls in longitudinal sections. Ano1 (green) was nicely present at endothelium and co-localized with endothelial marker (red) (Fig. 3C; the white arrow heads indicate blood vessels). Confocal imaging analyses also revealed that a co-localization of Ano1 (green) and CD31 (red) occurred at the plasma membrane of CVECs (Fig. 3D) because the highest merged fluorescence intensity (peaks) appeared in the vicinity of the plasma membrane (Fig. 3E). These results led us to hypothesize that the ICl(Ca) is carried by Ano1 in CVECs.

Bottom Line: The density of ICl ( C a) detected in CVECs was significantly inhibited by T16Ainh -A01, an Ano1 inhibitor, and a pore-targeting, specific anti-Ano1 antibody, and was markedly decreased in Ano1 gene knockdown CVECs.Ano1 formed CaCC in CVECs of neonatal mice.Ano1 may play a role in the pathophysiological processes during ischaemia in heart, and therefore, Ano1 might be a potential therapeutic target to prevent ischaemic damage.

View Article: PubMed Central - PubMed

Affiliation: Departments of Clinical Pharmacy and Cardiology, Institute of Clinical Pharmacy, The 2nd Affiliated Hospital, Key Laboratories of Education Ministry for Myocardial Ischemia Mechanism and Treatment, Harbin Medical University, Harbin, China.

No MeSH data available.


Related in: MedlinePlus