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Clusters of specialized detector cells provide sensitive and high fidelity receptor signaling in the intact endothelium.

Wilson C, Saunter CD, Girkin JM, McCarron JG - FASEB J. (2016)

Bottom Line: D., Girkin, J.M., McCarron, J.Clusters of specialized detector cells provide sensitive and high fidelity receptor signaling in the intact endothelium.

View Article: PubMed Central - PubMed

Affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, United Kingdom; and.

No MeSH data available.


Related in: MedlinePlus

ACh-evoked Ca2+ signals in endothelial cells of an intact and pressurized rat carotid artery. In response to ACh, a Ca2+ increase initiated in one part of the endothelium and progressed from there. A–C) Images at 1-s intervals show active wave fronts of Ca2+ release as the wave progressed across the endothelium. The wave fronts were determined by sequential subtraction of image frames. D) A color-coded representation of the time of activation of the entire population of cells (blue early, red late). The time color-code is at left of image. E, F) Grayscale representation of all cells exhibiting Ca2+ activity (E) with individual manually placed circular ROIs encompassing single cells (red circles) and around the entire field (orange circle) (F). Scale bars, 100 μm. G) Cellular (black) and averaged (orange/red) Ca2+ signals from the ROIs. H) Selected Ca2+ signals from (G) mapped to color-code shown in D. I) Plotting ACh-evoked Ca2+ signals (left) from ∼200 cells illustrates the temporal heterogeneity of Ca2+ responses. Global mean data (red line) represents the data poorly. Ca2+ signals were differentiated and then aligned in time (right) with respect to the peak of the derivative Ca2+ signal, to synchronize the Ca2+ signals in each cell and illustrate total Ca2+ activity (thick red line).
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Figure 2: ACh-evoked Ca2+ signals in endothelial cells of an intact and pressurized rat carotid artery. In response to ACh, a Ca2+ increase initiated in one part of the endothelium and progressed from there. A–C) Images at 1-s intervals show active wave fronts of Ca2+ release as the wave progressed across the endothelium. The wave fronts were determined by sequential subtraction of image frames. D) A color-coded representation of the time of activation of the entire population of cells (blue early, red late). The time color-code is at left of image. E, F) Grayscale representation of all cells exhibiting Ca2+ activity (E) with individual manually placed circular ROIs encompassing single cells (red circles) and around the entire field (orange circle) (F). Scale bars, 100 μm. G) Cellular (black) and averaged (orange/red) Ca2+ signals from the ROIs. H) Selected Ca2+ signals from (G) mapped to color-code shown in D. I) Plotting ACh-evoked Ca2+ signals (left) from ∼200 cells illustrates the temporal heterogeneity of Ca2+ responses. Global mean data (red line) represents the data poorly. Ca2+ signals were differentiated and then aligned in time (right) with respect to the peak of the derivative Ca2+ signal, to synchronize the Ca2+ signals in each cell and illustrate total Ca2+ activity (thick red line).

Mentions: Activation of the endothelium by extraluminal ACh [100 μM applied to the chamber, 100 nM estimated at the vessel lumen (see Supplemental Fig. 2); 60 mmHg Supplemental Movie 1)] evoked rises in [Ca2+]c in the majority of cells in the field (Fig. 2A–D). Within cells, repeating Ca2+ oscillations (uniform rises throughout the cell) and propagating Ca2+ waves (which moved at a velocity of 43 ± 3 µm/s; 60 cells from 6 arteries; not shown) occurred in various cells (Supplemental Movie 1). The Ca2+ rises evoked by ACh originated from an IP3-sensitive Ca2+ store. The Ca2+ increase persisted in a Ca2+-free bathing solution but was blocked by the sarco-endoplasmic reticulum Ca2+-ATPase inhibitor cyclopiazonic acid and the IP3 receptor blocker 2-aminoethoxydiphenyl borate (25). Caffeine failed to evoke a Ca2+ increase and ryanodine did not alter the ACh-evoked Ca2+ rise, suggesting ryanodine receptors play a minor role in Ca2+ signaling (25).


Clusters of specialized detector cells provide sensitive and high fidelity receptor signaling in the intact endothelium.

Wilson C, Saunter CD, Girkin JM, McCarron JG - FASEB J. (2016)

ACh-evoked Ca2+ signals in endothelial cells of an intact and pressurized rat carotid artery. In response to ACh, a Ca2+ increase initiated in one part of the endothelium and progressed from there. A–C) Images at 1-s intervals show active wave fronts of Ca2+ release as the wave progressed across the endothelium. The wave fronts were determined by sequential subtraction of image frames. D) A color-coded representation of the time of activation of the entire population of cells (blue early, red late). The time color-code is at left of image. E, F) Grayscale representation of all cells exhibiting Ca2+ activity (E) with individual manually placed circular ROIs encompassing single cells (red circles) and around the entire field (orange circle) (F). Scale bars, 100 μm. G) Cellular (black) and averaged (orange/red) Ca2+ signals from the ROIs. H) Selected Ca2+ signals from (G) mapped to color-code shown in D. I) Plotting ACh-evoked Ca2+ signals (left) from ∼200 cells illustrates the temporal heterogeneity of Ca2+ responses. Global mean data (red line) represents the data poorly. Ca2+ signals were differentiated and then aligned in time (right) with respect to the peak of the derivative Ca2+ signal, to synchronize the Ca2+ signals in each cell and illustrate total Ca2+ activity (thick red line).
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Related In: Results  -  Collection

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Show All Figures
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Figure 2: ACh-evoked Ca2+ signals in endothelial cells of an intact and pressurized rat carotid artery. In response to ACh, a Ca2+ increase initiated in one part of the endothelium and progressed from there. A–C) Images at 1-s intervals show active wave fronts of Ca2+ release as the wave progressed across the endothelium. The wave fronts were determined by sequential subtraction of image frames. D) A color-coded representation of the time of activation of the entire population of cells (blue early, red late). The time color-code is at left of image. E, F) Grayscale representation of all cells exhibiting Ca2+ activity (E) with individual manually placed circular ROIs encompassing single cells (red circles) and around the entire field (orange circle) (F). Scale bars, 100 μm. G) Cellular (black) and averaged (orange/red) Ca2+ signals from the ROIs. H) Selected Ca2+ signals from (G) mapped to color-code shown in D. I) Plotting ACh-evoked Ca2+ signals (left) from ∼200 cells illustrates the temporal heterogeneity of Ca2+ responses. Global mean data (red line) represents the data poorly. Ca2+ signals were differentiated and then aligned in time (right) with respect to the peak of the derivative Ca2+ signal, to synchronize the Ca2+ signals in each cell and illustrate total Ca2+ activity (thick red line).
Mentions: Activation of the endothelium by extraluminal ACh [100 μM applied to the chamber, 100 nM estimated at the vessel lumen (see Supplemental Fig. 2); 60 mmHg Supplemental Movie 1)] evoked rises in [Ca2+]c in the majority of cells in the field (Fig. 2A–D). Within cells, repeating Ca2+ oscillations (uniform rises throughout the cell) and propagating Ca2+ waves (which moved at a velocity of 43 ± 3 µm/s; 60 cells from 6 arteries; not shown) occurred in various cells (Supplemental Movie 1). The Ca2+ rises evoked by ACh originated from an IP3-sensitive Ca2+ store. The Ca2+ increase persisted in a Ca2+-free bathing solution but was blocked by the sarco-endoplasmic reticulum Ca2+-ATPase inhibitor cyclopiazonic acid and the IP3 receptor blocker 2-aminoethoxydiphenyl borate (25). Caffeine failed to evoke a Ca2+ increase and ryanodine did not alter the ACh-evoked Ca2+ rise, suggesting ryanodine receptors play a minor role in Ca2+ signaling (25).

Bottom Line: D., Girkin, J.M., McCarron, J.Clusters of specialized detector cells provide sensitive and high fidelity receptor signaling in the intact endothelium.

View Article: PubMed Central - PubMed

Affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, United Kingdom; and.

No MeSH data available.


Related in: MedlinePlus