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Effect of the Epac activator 8‐pCPT‐2′‐O‐Me‐cAMP (8‐pCPT) on the expression pattern of VE‐cadherin (VE‐cad) & connexins (Cxs) in Human Coronary Artery Endothelial Cells (HCAECs)
Author(s) -
Quinn Rachael Ellen,
Roberts Owain,
Kamishima Tomoko,
Dart Caroline,
Quayle John
Publication year - 2013
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.27.1_supplement.707.7
Subject(s) - adherens junction , cadherin , gap junction , biology , microbiology and biotechnology , activator (genetics) , medicine , cell junction , endocrinology , chemistry , cell , intracellular , biochemistry , gene
Homocellular junctions between endothelial cells include adherens junctions, formed by cadherins, & gap junctions, formed by Cxs. Dynamic regulation of these junctions is necessary to maintain the endothelial barrier. We examined the effect of Ca 2+ & 8‐pCPT, a selective activator of e xchange p rotein directly a ctivated by c AMP (Epac), on the expression pattern of VE‐cad & Cxs in HCAECs. VE‐cad, Cx37, 40 & 43 were identified in HCAECs by immunocytochemistry. VE‐cad was identified in parts of the cell membrane, without overlapping with Cx37, 40 or 43. Application of 8‐pCPT produced more continuous VE‐cad membrane staining that overlapped with Cx37. Disruption of VE‐cad interactions using VE‐cad antibody changed HCAEC morphology from flattened & continuous to small dispersed colonies. Removal of Ca 2+ from medium resulted in rounded cells isolated from each other. Following Ca 2+ restoration, a normal pattern of VE‐cad membrane staining returned, followed by return of membrane Cx staining. Addition of VE‐cad antibody blocked this recovery. Epac activation promotes localisation of VE‐cad to the membrane & adherens junction formation. Redistribution of VE‐cad has a subsequent effect on Cx localisation. Adherens junction formation in HCAECs therefore precedes & promotes Cx interactions at cell junctions. Funded by the University of Liverpool