Purchase this article with an account.
C. M. Hutnik, S. Khimdas, H. Liu, C. Pocrnich, D. Laird, C. Shao; Inhibitory Effect of Cx43 on Oxidative Stress Induced VEGF in Human Retinal Pigment Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2010;51(13):1433.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
To determine the effect of connexin43 (Cx43) expression and gap junctional intercellular communication (GJIC) on the expression and secretion of vascular endothelial growth factor (VEGF) from the retinal pigment epithelium under normal cell culture and oxidative stress conditions.
Stable cell lines of ARPE-19 were produced in which Cx43 was either over-expressed, down-regulated by targeted shRNA, or functionally inhibited by co-expression of a disease-linked dominant-negative mutant (G21R). Pharmacologic blockade of GJIC was accomplished with flufenamic acid. Oxidant challenge was performed with tert-butyl hydroperoxide (tBH). VEGF gene expression and secretion were assessed by real-time PCR and ELISA, respectively. Serum was collected from wild-type mice, mice expressing a dominant-negative mutant of Cx43 and Cx43 null mice.
Over-expression of Cx43 in ARPE-19 cells reduced both gene expression and secretion of VEGF. Down-regulation of Cx43 increased gene expression and secretion of VEGF. Increased secretion of VEGF was also observed in ARPE-19 cells expressing a dominant negative mutant of Cx43 or when GJIC was blocked. Over-expression of Cx43 reduced tBH-induced secretion of VEGF from ARPE-19 cells. Finally, Cx43 mutant mice and Cx43 knock-out mice displayed relatively higher levels of serum VEGF than wild-type mice.
Expression of Cx43 in ARPE-19 cells reduces VEGF secretion under normal cell culture conditions and protects against tBH-induced VEGF secretion.
This PDF is available to Subscribers Only