Unique gene expression profiles of donor-matched human retinal and choroidal vascular endothelial cells.
OBJECTIVE
Consistent with clinical observations that posterior uveitis frequently involves the retinal vasculature and recent recognition of vascular heterogeneity, the hypothesis for this study was that retinal vascular endothelium was a cell population of unique molecular phenotype.


METHODS
Donor-matched cultures of primary retinal and choroidal endothelial cells from six human cadavers were incubated with either Toxoplasma gondii tachyzoites (10:1, parasites per cell) or Escherichia coli lipopolysaccharide (100 ng/mL); control cultures were simultaneously incubated with medium. Gene expression profiling of endothelial cells was performed using oligonucleotide arrays containing probes designed to detect 8746 human transcripts. After normalization, differential gene expression was assessed by the significance analysis of microarrays, with the false-discovery rate set at 5%. For selected genes, differences in the level of expression between retinal and choroidal cells were evaluated by real-time RT-PCR.


RESULTS
Graphic descriptive analysis demonstrated a strong correlation between gene expression of unstimulated retinal and choroidal endothelial cells, but also highlighted distinctly different patterns of expression that were greater than differences noted between donors or between unstimulated and stimulated cells. Overall, 779 (8.9%) of 8746 transcripts were differentially represented. Of note, the 330 transcripts that were present at higher levels in retinal cells included a larger percentage of transcripts encoding molecules involved in the immune response. Differential gene expression was confirmed for 12 transcripts by RT-PCR.


CONCLUSIONS
Retinal and choroidal vascular endothelial cells display distinctive gene expression profiles. The findings suggest the possibility of treating posterior uveitis by targeting specific interactions between the retinal endothelial cell and an infiltrating leukocyte.
Smith
Justine R
JR
Casey Eye Institute, Oregon Health and Science University, Portland, Oregon 97239, USA. smithjus@ohsu.edu
Choi
Dongseok
D
Chipps
Timothy J
TJ
Pan
Yuzhen
Y
Zamora
David O
DO
Davies
Michael H
MH
Babra
Bobby
B
Powers
Michael R
MR
Planck
Stephen R
SR
Rosenbaum
James T
JT
D000328:Adult; D000818:Animals; D002675:Child, Preschool; D002829:Choroid / Q000098:blood supply*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism* / Q000469:parasitology; D004926:Escherichia coli; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D012171:Retinal Vessels / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014019:Tissue Donors; D014122:Toxoplasma / Q000502:physiology
D008070:Lipopolysaccharides