April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Increased expression of inflammatory markers in the RPE of CfhTg mice and in RD8+/+ mice.
Author Affiliations & Notes
  • Cynthia Xin-Zhao Wang
    Ophthalmology, UT Southwerstern Medical Center, Dallas, TX
  • Bogale Aredo
    Ophthalmology, UT Southwerstern Medical Center, Dallas, TX
  • Tao Li
    Ophthalmology, UT Southwerstern Medical Center, Dallas, TX
  • Rafael Ufret-Vincenty
    Ophthalmology, UT Southwerstern Medical Center, Dallas, TX
  • Footnotes
    Commercial Relationships Cynthia Wang, None; Bogale Aredo, None; Tao Li, None; Rafael Ufret-Vincenty, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 3280. doi:https://doi.org/
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      Cynthia Xin-Zhao Wang, Bogale Aredo, Tao Li, Rafael Ufret-Vincenty; Increased expression of inflammatory markers in the RPE of CfhTg mice and in RD8+/+ mice.. Invest. Ophthalmol. Vis. Sci. 2014;55(13):3280. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: Our aim is to determine how the expression of genes by the RPE is altered by two different factors: 1. the 402H variant of complement factor H, and 2. the RD8 mutation of the Crb1 gene.

Methods: Using the SRIRS (Simultaneous RPE cell Isolation and RNA Stabilization) technique, we isolated RPE cells from mouse eyecups of 2 year old CfhTg/mCfhKO and C57BL/6J mice, and isolated RNA from these RPE cells. We ran Illumina microarray chips on these samples, and performed a pathway analysis on the resulting genes. QPCR was used to test a subset of these genes and also an independent group of candidate genes for qPCR analysis. We also isolated RNA from the RPE (and also the retina separately) of 8m RD8+/+ vs. B6-wild type (WT) mice and performed qPCR for candidate genes.

Results: We were able to isolate enough RNA from the RPE layer (and overlying subretinal microglia) from individual eyecups to perform Illumina and qPCR testing. QPCR confirmed Illumina findings: an increased expression of CD68 and TREM2, and a decreased expression of SNCA in CfhTg mice vs. B6 controls. Some candidate genes were also found to be increased in CfhTg/mCfhKO mice including: IP-10, NLRP3 and C4. In RD8 +/+ mice, qPCR of candidate genes revealed an increased expression of Ccl-2, CFB, C3, NF-kB and CD200R in the RPE compared to WT mice. The retina of RD8 +/+ mice showed an increased expression of oxidative stress marker HO-1, and complement components C1q and C4 compared to WT.

Conclusions: The 402H variant of Cfh can lead to an increased expression of inflammatory markers in the RPE. The RD8 mutation, which causes a retinal degeneration, also leads to a distinct increase in inflammatory gene expression in the RPE, and an increase in oxidative stress and complement activation markers in the retina.

Keywords: 412 age-related macular degeneration • 539 genetics • 533 gene/expression  
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