Changes in retinal pigment epithelial gene expression induced by rod outer segment uptake

Itay Chowers, Yoonhee Kim, Ronald H. Farkas, Tushara L. Gunatilaka, Abigail S Hackam, Peter A. Campochiaro, Silvia C. Finnemann, Donald J. Zack

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

PURPOSE. Rod outer segment (ROS) uptake, a crucial function of the retinal pigment epithelium (RPE), probably involve multiple proteins, yet only a small number have so far been identified. The goal of this study was to find additional genes involved in ROS uptake and degradation by identifying ROS-induced gene expression changes. METHODS. Human RPE-derived ARPE-19 cells were harvested 3 and 12 hours after addition of bovine ROS. Gene expression profiles were compared with control cultures by using a custom human retina cDNA microarray and were validated by quantitative real-time RT-PCR (QPCR). ROS binding and internalization were quantitated with a fluorescence assay. RESULTS. Alterations in the expression levels of multiple genes (especially ones involved in transcriptional regulation, signal transduction, or protein modification) were detected 3 hours after ROS challenge, whereas by 12 hours most had returned to baseline. QPCR results corroborated the microarray results for seven of the eight genes tested. Time-course QPCR experiments on an independent sample set demonstrated characteristic temporal expression changes for each gene. Protein levels of one of these, plasminogen activator inhibitor-1 (PAI-1), were tested and found to parallel the mRNA changes. In addition, exogenous PAI-1 inhibited ROS uptake by RPE cells in vitro, consistent with its putative function in integrin receptor regulation. CONCLUSIONS. ROS uptake is associated with regulation of multiple RPE genes in a gene-specific temporal pattern. These genes are candidates for involvement in ROS uptake and degradation, particularly PAI-1, for which the study provided evidence suggesting that it may participate in the negative feedback of ROS uptake.

Original languageEnglish
Pages (from-to)2098-2106
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume45
Issue number7
DOIs
StatePublished - Jul 1 2004
Externally publishedYes

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Rod Cell Outer Segment
Retinal Pigments
Gene Expression
Retinal Pigment Epithelium
Plasminogen Activator Inhibitor 1
Genes
Proteins
Oligonucleotide Array Sequence Analysis
Transcriptome
Integrins
Retina
Real-Time Polymerase Chain Reaction
Signal Transduction
Fluorescence

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Changes in retinal pigment epithelial gene expression induced by rod outer segment uptake. / Chowers, Itay; Kim, Yoonhee; Farkas, Ronald H.; Gunatilaka, Tushara L.; Hackam, Abigail S; Campochiaro, Peter A.; Finnemann, Silvia C.; Zack, Donald J.

In: Investigative Ophthalmology and Visual Science, Vol. 45, No. 7, 01.07.2004, p. 2098-2106.

Research output: Contribution to journalArticle

Chowers, I, Kim, Y, Farkas, RH, Gunatilaka, TL, Hackam, AS, Campochiaro, PA, Finnemann, SC & Zack, DJ 2004, 'Changes in retinal pigment epithelial gene expression induced by rod outer segment uptake', Investigative Ophthalmology and Visual Science, vol. 45, no. 7, pp. 2098-2106. https://doi.org/10.1167/iovs.03-0863
Chowers, Itay ; Kim, Yoonhee ; Farkas, Ronald H. ; Gunatilaka, Tushara L. ; Hackam, Abigail S ; Campochiaro, Peter A. ; Finnemann, Silvia C. ; Zack, Donald J. / Changes in retinal pigment epithelial gene expression induced by rod outer segment uptake. In: Investigative Ophthalmology and Visual Science. 2004 ; Vol. 45, No. 7. pp. 2098-2106.
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