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Disruption of spectrin‐f‐actin binding is sufficient to induce inter‐endothelial gaps
Author(s) -
Prasain Nutan,
Wu Songwei,
Goodman Steven R.,
Stevens Troy
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a748-a
Subject(s) - spectrin , microbiology and biotechnology , cytoskeleton , actin , actin cytoskeleton , endothelial stem cell , biology , transfection , epb41 , actina , actin binding protein , chemistry , cell , cell culture , in vitro , biochemistry , genetics
Spectrin is an essential component of the membrane‐associated cytoskeleton that directly cross‐links f‐actin via its actin binding domain (ABD). While f‐actin re‐aligns from a cortical distribution into stress fibers necessary for inter‐endothelial gap formation, it is unclear whether disruption of f‐actin‐spectrin association in the membrane cytoskeleton is sufficient to induce inter‐endothelial gaps. We hypothesized that spectrin‐f‐actin binding is essential for maintaining endothelial cell shape and that its disruption is sufficient to induce inter‐endothelial gaps. We used the Chariot transfection system to deliver SG 43 antibody into confluent monolayers of both pulmonary artery endothelial cells (PAECs) and pulmonary microvascular endothelial cells (PMVECs). SG 43 targets a region adjacent to the ABD of β‐spectrin and selectively dislodges actin from its binding with spectrin. SG 43 induced inter‐endothelial gaps in PAECs, whereas in PMVECs it induced only transient or minor gaps. Thapsigargin (1μM) did not induce gaps in PMVECs, although SG 43 treatment revealed thapsigargin‐induced gap formation in the microvascular endothelial cells. Collectively, these data suggest that disruption of the spectrin‐f‐actin interaction is sufficient to induce gaps in PAECs, but not in PMVECs, suggesting that the PAECs are particularly reliant on the spectrin membrane skeleton for regulation of their shape. Supported by HL66299 and HL60024.