The role of Bcl-xL in mouse RPE cell survival.

Published online

Journal Article

PURPOSE. Retinal pigment epithelial (RPE) cell survival plays a critical role in normal physiology and in retinal diseases, such as age-related macular degeneration (AMD) and proliferative vitreoretinopathy (PVR). We have previously demonstrated that Bcl-x(L) is an important cell survival protein in human RPE (hRPE) cells. Herein, we determined the role of Bcl-x(L) as a survival protein in mouse RPE (mRPE) cells. METHODS. Survival factor gene expression and Bcl-x(L) protein distribution were determined using qRT-PCR and immunohistochemistry, respectively. Cultured mRPE cells were transfected with two modified 2'-O-methoxyethoxy antisense oligonucleotides (ASOs): Bcl-x(L)-mismatched control and Bcl-x(L)-specific. Bcl-x(L) protein levels were analyzed using Western blot. To determine the effects of survival factor regulation in mRPE cells, cultured cells were treated for 24 hours with mouse TNF-α, human IL-1β, and human TNF-α. RESULTS. Bcl-x(L) was the most highly expressed survival factor in both mouse eyecup and cultured mRPE cells, whereas Bax was the most highly expressed antisurvival factor. Bcl-x(L) was expressed in the RPE layer, and the distribution among the retinal layers was similar to that observed in human eyecups. IL-1β and TNF-α had minimal effect on Bcl-x(L) and Bax expression and strongly upregulated Traf-1. Transfection with Bcl-x(L)-specific ASO resulted in markedly diminished Bcl-x(L) gene expression, Bcl-x(L) protein levels, and cell number. CONCLUSIONS. Bcl-x(L) is the most highly expressed survival gene in mRPE cells and is essential for mRPE cell survival. Our data suggest that mouse tissue is an appropriate model for investigations of RPE survival factor genes.

Full Text

Duke Authors

Cited Authors

  • Medearis, S; Han, IC; Huang, JK; Yang, P; Jaffe, GJ

Published Date

  • August 17, 2011

Published In

Volume / Issue

  • 52 / 9

Start / End Page

  • 6545 - 6551

PubMed ID

  • 21724914

Pubmed Central ID

  • 21724914

Electronic International Standard Serial Number (EISSN)

  • 1552-5783

Digital Object Identifier (DOI)

  • 10.1167/iovs.10-6772

Language

  • eng

Conference Location

  • United States