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Andrew T C Tsin, Robert Moritz, Sandeep Vellanki, Richard LeBaron; Integrin Expression in Cultured Retinal Endothelial Cells and Retinal Pericytes. Invest. Ophthalmol. Vis. Sci. 2016;57(12):3233.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: BIGH3, an ECM-proapoptotic protein has been shown to induce apoptosis in Rhesus Monkey Retinal Endothelial Cells (RhREC) and Primary Human Retinal Pericytes (HRP). The C-terminus of BIGH3 has two peptide sequences EPDIM and RGD. These sequences interact with integrin receptors laminin (α3β1) and fibronectin (α5β1), respectively, and can induce a BIGH3-mediated apoptotic response. The objective of current study is to investigate the extent that RhREC and HRP express α3β1 and α5β1 in cell culture conditions.
Methods: cDNA generated from total RNA was isolated from HRP and RhREC. Quantitative PCR was performed on the cDNA to measure α3 and α5 subunit expression. 18s rRNA was used as internal control. Human Renal Proximal Tubule Cells (RPTEC) serves as control to determine fold change.
Results: In the cell conditions tested, we did not detect integrin subunit α3 expression in RhREC. Compared to RPTEC the fold change of α3 in HRP was 0.1. Integrin subunit α5 mRNA was expressed by both RhREC and HRP. The fold change was 0.004 in REC and 0.95 in HRP when values were compared to RPTEC.
Conclusions: These data indicate that RhREC expressed little, if any α3 subunit mRNA, but did express α5β1. In contrast, HRP expressed both α3β1 and α5β1 integrins in the tested conditions. Differential integrin expression in RhREC and HRP indicates that these cells show distinctive responses to BIGH3-mediated apoptosis. The roles of α3β1 and α5β1 integrins in in BIGH3 mediated apoptosis of retinal cells will be further investigated.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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