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Endothelial cells use dynamic actin to facilitate lymphocyte transendothelial migration and maintain the monolayer barrier.

Mooren OL, Li J, Nawas J, Cooper JA - Mol. Biol. Cell (2014)

Bottom Line: The actin cytoskeleton of the endothelial cell (EC) is known to facilitate transmigration, but the cellular and molecular mechanisms are not well understood.We found that docking structure formation involves the localization and activation of Arp2/3 complex by WAVE2.Finally, we found that ECs in resting endothelial monolayers use lamellipodial protrusions dependent on WAVE2 to form and maintain contacts and junctions between cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology and Physiology, Washington University, St. Louis, MO 63110.

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WAVE2 is important for junction assembly after calcium depletion and addition. (A) Calcium was removed from the medium with EGTA to disassemble cadherin-based junctions and then replaced to induce reassembly. Images are representative of 5 fov from each of three samples. Scale bar, 50 μm. (B) WAVE2 knockdown causes a defect in junctional assembly after calcium addition, based on TER measurements over time. (C) Time course of junction assembly revealed by the TER data from B normalized for control and WAVE2-depleted monolayers. The time courses are similar, suggesting that junctions recover at a normal rate.
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Figure 7: WAVE2 is important for junction assembly after calcium depletion and addition. (A) Calcium was removed from the medium with EGTA to disassemble cadherin-based junctions and then replaced to induce reassembly. Images are representative of 5 fov from each of three samples. Scale bar, 50 μm. (B) WAVE2 knockdown causes a defect in junctional assembly after calcium addition, based on TER measurements over time. (C) Time course of junction assembly revealed by the TER data from B normalized for control and WAVE2-depleted monolayers. The time courses are similar, suggesting that junctions recover at a normal rate.

Mentions: To investigate further the role of WAVE2 in cell junction formation, we challenged EC monolayers to assemble junctions by Ca2+ depletion and addition (Figure 7, Supplemental Figure S4, and Table 1). VE-cadherin–based junctions are lost when Ca2+ is removed, and they assemble when Ca2+ is added back. We found that depletion of WAVE2 impaired the efficiency of ECs to form cell junctions in this assay (Figure 7, Supplemental Figure S4, and Table 1). At all time points during reassembly, the number of junctions in WAVE2-depleted monolayers was lower than in control monolayers (Figure 7, Supplemental Figure S4, and Table 1). When junctions re-formed in the WAVE2-depleted monolayers, the fluorescence intensity of the tight-junction protein ZO-1 was similar to that in control monolayers (Figure 7 and Table 1).


Endothelial cells use dynamic actin to facilitate lymphocyte transendothelial migration and maintain the monolayer barrier.

Mooren OL, Li J, Nawas J, Cooper JA - Mol. Biol. Cell (2014)

WAVE2 is important for junction assembly after calcium depletion and addition. (A) Calcium was removed from the medium with EGTA to disassemble cadherin-based junctions and then replaced to induce reassembly. Images are representative of 5 fov from each of three samples. Scale bar, 50 μm. (B) WAVE2 knockdown causes a defect in junctional assembly after calcium addition, based on TER measurements over time. (C) Time course of junction assembly revealed by the TER data from B normalized for control and WAVE2-depleted monolayers. The time courses are similar, suggesting that junctions recover at a normal rate.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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Figure 7: WAVE2 is important for junction assembly after calcium depletion and addition. (A) Calcium was removed from the medium with EGTA to disassemble cadherin-based junctions and then replaced to induce reassembly. Images are representative of 5 fov from each of three samples. Scale bar, 50 μm. (B) WAVE2 knockdown causes a defect in junctional assembly after calcium addition, based on TER measurements over time. (C) Time course of junction assembly revealed by the TER data from B normalized for control and WAVE2-depleted monolayers. The time courses are similar, suggesting that junctions recover at a normal rate.
Mentions: To investigate further the role of WAVE2 in cell junction formation, we challenged EC monolayers to assemble junctions by Ca2+ depletion and addition (Figure 7, Supplemental Figure S4, and Table 1). VE-cadherin–based junctions are lost when Ca2+ is removed, and they assemble when Ca2+ is added back. We found that depletion of WAVE2 impaired the efficiency of ECs to form cell junctions in this assay (Figure 7, Supplemental Figure S4, and Table 1). At all time points during reassembly, the number of junctions in WAVE2-depleted monolayers was lower than in control monolayers (Figure 7, Supplemental Figure S4, and Table 1). When junctions re-formed in the WAVE2-depleted monolayers, the fluorescence intensity of the tight-junction protein ZO-1 was similar to that in control monolayers (Figure 7 and Table 1).

Bottom Line: The actin cytoskeleton of the endothelial cell (EC) is known to facilitate transmigration, but the cellular and molecular mechanisms are not well understood.We found that docking structure formation involves the localization and activation of Arp2/3 complex by WAVE2.Finally, we found that ECs in resting endothelial monolayers use lamellipodial protrusions dependent on WAVE2 to form and maintain contacts and junctions between cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology and Physiology, Washington University, St. Louis, MO 63110.

Show MeSH
Related in: MedlinePlus