Limits...
Distribution of cytoskeletal components in endothelial cells in the Guinea pig renal artery.

Katoh K, Noda Y - Int J Cell Biol (2012)

Bottom Line: Renal arterial endothelial cells were shown to be elongated along the direction of blood flow, while stress fibers ran perpendicular to the flow in the basal portion.On the other hand, stress fibers in the apical portion of cells ran along the direction of flow.These observations suggest that the morphological alignment of endothelial cells along the direction of blood flow and the organization of cytoskeletal components are independently regulated.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Jichi Medical School, 3311-1 Yakushiji, Shimotsuke, Tochigi 329-0498, Japan.

ABSTRACT
The cytoskeletal components of endothelial cells in the renal artery were examined by analysis of en face preparations under confocal laser scanning microscopy. Renal arterial endothelial cells were shown to be elongated along the direction of blood flow, while stress fibers ran perpendicular to the flow in the basal portion. Focal adhesions were observed along the stress fibers in dot-like configurations. On the other hand, stress fibers in the apical portion of cells ran along the direction of flow. The localizations of stress fibers and focal adhesions in endothelial cells in the renal artery differed from those of unperturbed aortic and venous endothelial cells. Tyrosine-phosphorylated proteins were mainly detected at the sites of cell-to-cell apposition, but not in focal adhesions. Pulsatile pressure and fluid shear stress applied over endothelial cells in the renal artery induce stress fiber organization and localization of focal adhesions. These observations suggest that the morphological alignment of endothelial cells along the direction of blood flow and the organization of cytoskeletal components are independently regulated.

No MeSH data available.


Related in: MedlinePlus

Localization of tyrosine-phosphorylated proteins and vinculin in the renal arterial endothelial cells. En face preparations were stained with antiphosphotyrosine antibody (PY-20) (a–c) or antivinculin antibody (d–f) and observed by confocal laser scanning microscopy. The focal plane was adjusted to the surface (a and d), middle (b and e), or basal (c and f) portion of the endothelial cells. Tyrosine-phosphorylated proteins were observed along the apical plasma membrane in uniform staining (a). Strong staining for tyrosine-phosphorylated proteins was detected at sites of cell-to-cell apposition (b), with faint staining in the cytoplasm (c). Antivinculin staining was detected at sites of cell-to-cell apposition (e) and focal adhesion in the basal portion of the cells (f: arrowheads). No staining for vinculin was detected at the apical surface of the cells (d). The arrow indicates the direction of blood flow. Bar: 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3303860&req=5

fig3: Localization of tyrosine-phosphorylated proteins and vinculin in the renal arterial endothelial cells. En face preparations were stained with antiphosphotyrosine antibody (PY-20) (a–c) or antivinculin antibody (d–f) and observed by confocal laser scanning microscopy. The focal plane was adjusted to the surface (a and d), middle (b and e), or basal (c and f) portion of the endothelial cells. Tyrosine-phosphorylated proteins were observed along the apical plasma membrane in uniform staining (a). Strong staining for tyrosine-phosphorylated proteins was detected at sites of cell-to-cell apposition (b), with faint staining in the cytoplasm (c). Antivinculin staining was detected at sites of cell-to-cell apposition (e) and focal adhesion in the basal portion of the cells (f: arrowheads). No staining for vinculin was detected at the apical surface of the cells (d). The arrow indicates the direction of blood flow. Bar: 50 μm.

Mentions: As certain proteins undergo tyrosine phosphorylation during intracellular signaling events, we next examined the tyrosine phosphorylation levels of endothelial cells in the renal artery (Figure 3). En face preparations stained with an antibody against phosphotyrosine-containing proteins or an antibody against vinculin were examined by confocal laser scanning microscopy. Positive staining with antiphosphotyrosine antibody was observed in a uniform pattern at the apical surface of the endothelial cells (Figure 3(a)). When the focal plane was adjusted to the middle level of the cells, staining was detected at sites of cell-to-cell apposition (Figure 3(b)). The focal plane was adjusted to the basal portion of the cell, and staining with antiphosphotyrosine antibody was mainly detected at cell-to-cell adhesion sites, but not along stress fibers or focal adhesions (Figure 3(c)). On the other hand, staining with antivinculin was detected at both sites of cell-to-cell apposition (Figure 3(e)) and focal adhesions at the basal side of the cell (Figure 3(f)). No significant staining was detected at the apical surface of the cell (Figure 3(d)). Staining with antivinculin and antiphosphotyrosine antibodies did not show complete colocalization at the basal portion of cells (Figures 3(c) and 3(f)). The renal artery was also double-stained for F-actin with rhodamine-labeled phalloidin and with antiphosphotyrosine antibody. Figure 4 shows a stereo view of a cell double-stained with rhodamine-labeled phalloidin to visualize actin filaments and with antiphosphotyrosine antibody, and the results indicated no colocalization of tyrosine-phosphorylated proteins and stress fibers. Positive staining for phosphotyrosine was detected on the plasma membrane over endothelial cells, especially along the cell-to-cell adhesion sites. However, the specific distribution of staining for phosphotyrosine was not detected along the stress fibers in the basal or apical portions of the cell.


Distribution of cytoskeletal components in endothelial cells in the Guinea pig renal artery.

Katoh K, Noda Y - Int J Cell Biol (2012)

Localization of tyrosine-phosphorylated proteins and vinculin in the renal arterial endothelial cells. En face preparations were stained with antiphosphotyrosine antibody (PY-20) (a–c) or antivinculin antibody (d–f) and observed by confocal laser scanning microscopy. The focal plane was adjusted to the surface (a and d), middle (b and e), or basal (c and f) portion of the endothelial cells. Tyrosine-phosphorylated proteins were observed along the apical plasma membrane in uniform staining (a). Strong staining for tyrosine-phosphorylated proteins was detected at sites of cell-to-cell apposition (b), with faint staining in the cytoplasm (c). Antivinculin staining was detected at sites of cell-to-cell apposition (e) and focal adhesion in the basal portion of the cells (f: arrowheads). No staining for vinculin was detected at the apical surface of the cells (d). The arrow indicates the direction of blood flow. Bar: 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3303860&req=5

fig3: Localization of tyrosine-phosphorylated proteins and vinculin in the renal arterial endothelial cells. En face preparations were stained with antiphosphotyrosine antibody (PY-20) (a–c) or antivinculin antibody (d–f) and observed by confocal laser scanning microscopy. The focal plane was adjusted to the surface (a and d), middle (b and e), or basal (c and f) portion of the endothelial cells. Tyrosine-phosphorylated proteins were observed along the apical plasma membrane in uniform staining (a). Strong staining for tyrosine-phosphorylated proteins was detected at sites of cell-to-cell apposition (b), with faint staining in the cytoplasm (c). Antivinculin staining was detected at sites of cell-to-cell apposition (e) and focal adhesion in the basal portion of the cells (f: arrowheads). No staining for vinculin was detected at the apical surface of the cells (d). The arrow indicates the direction of blood flow. Bar: 50 μm.
Mentions: As certain proteins undergo tyrosine phosphorylation during intracellular signaling events, we next examined the tyrosine phosphorylation levels of endothelial cells in the renal artery (Figure 3). En face preparations stained with an antibody against phosphotyrosine-containing proteins or an antibody against vinculin were examined by confocal laser scanning microscopy. Positive staining with antiphosphotyrosine antibody was observed in a uniform pattern at the apical surface of the endothelial cells (Figure 3(a)). When the focal plane was adjusted to the middle level of the cells, staining was detected at sites of cell-to-cell apposition (Figure 3(b)). The focal plane was adjusted to the basal portion of the cell, and staining with antiphosphotyrosine antibody was mainly detected at cell-to-cell adhesion sites, but not along stress fibers or focal adhesions (Figure 3(c)). On the other hand, staining with antivinculin was detected at both sites of cell-to-cell apposition (Figure 3(e)) and focal adhesions at the basal side of the cell (Figure 3(f)). No significant staining was detected at the apical surface of the cell (Figure 3(d)). Staining with antivinculin and antiphosphotyrosine antibodies did not show complete colocalization at the basal portion of cells (Figures 3(c) and 3(f)). The renal artery was also double-stained for F-actin with rhodamine-labeled phalloidin and with antiphosphotyrosine antibody. Figure 4 shows a stereo view of a cell double-stained with rhodamine-labeled phalloidin to visualize actin filaments and with antiphosphotyrosine antibody, and the results indicated no colocalization of tyrosine-phosphorylated proteins and stress fibers. Positive staining for phosphotyrosine was detected on the plasma membrane over endothelial cells, especially along the cell-to-cell adhesion sites. However, the specific distribution of staining for phosphotyrosine was not detected along the stress fibers in the basal or apical portions of the cell.

Bottom Line: Renal arterial endothelial cells were shown to be elongated along the direction of blood flow, while stress fibers ran perpendicular to the flow in the basal portion.On the other hand, stress fibers in the apical portion of cells ran along the direction of flow.These observations suggest that the morphological alignment of endothelial cells along the direction of blood flow and the organization of cytoskeletal components are independently regulated.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Jichi Medical School, 3311-1 Yakushiji, Shimotsuke, Tochigi 329-0498, Japan.

ABSTRACT
The cytoskeletal components of endothelial cells in the renal artery were examined by analysis of en face preparations under confocal laser scanning microscopy. Renal arterial endothelial cells were shown to be elongated along the direction of blood flow, while stress fibers ran perpendicular to the flow in the basal portion. Focal adhesions were observed along the stress fibers in dot-like configurations. On the other hand, stress fibers in the apical portion of cells ran along the direction of flow. The localizations of stress fibers and focal adhesions in endothelial cells in the renal artery differed from those of unperturbed aortic and venous endothelial cells. Tyrosine-phosphorylated proteins were mainly detected at the sites of cell-to-cell apposition, but not in focal adhesions. Pulsatile pressure and fluid shear stress applied over endothelial cells in the renal artery induce stress fiber organization and localization of focal adhesions. These observations suggest that the morphological alignment of endothelial cells along the direction of blood flow and the organization of cytoskeletal components are independently regulated.

No MeSH data available.


Related in: MedlinePlus