Microarray analysis of choroid/RPE gene expression in marmoset eyes undergoing changes in ocular growth and refraction.
OBJECTIVE
Visually guided ocular growth is facilitated by scleral extracellular matrix remodeling at the posterior pole of the eye. Coincident with scleral remodeling, significant changes in choroidal morphology, blood flow, and protein synthesis have been shown to occur in eyes undergoing ocular growth changes. The current study is designed to identify gene expression changes that may occur in the choroid/retinal pigment epithelium (RPE) of marmoset eyes during their compensation for hyperopic defocus as compared to eyes compensating for myopic defocus.


METHODS
Total RNA was isolated from choroid/RPE from four common marmosets (Callithrix jacchus) undergoing binocular lens treatment using extended wear soft contact lenses of equal magnitude but opposite sign (+/-5 diopter [D]). After reverse transcription, cDNA was labeled and hybridized to a human oligonucleotide microarray and gene transcript expression profiles were determined. Real-time polymerase chain reaction (PCR) and western blot analysis were used to confirm genes and proteins of interest, respectively.


RESULTS
Microarray analyses in choroid/RPE indicated 204 genes were significantly changed in minus lens-treated as compared with plus lens-treated eyes (p<0.05, Student's t-test). Differential choroid/RPE expression of protein tyrosine phosphatase, receptor type, B (PTPRB), transforming growth factor beta-induced (TGFBI), and basic fibroblast growth factor 2 (FGF-2) were confirmed by real-time PCR. TGFBIp was confirmed at the protein level by western blot analysis in marmoset and human cornea, choroid/RPE, and sclera.


CONCLUSIONS
The present study demonstrated that significant gene expression changes occur in the marmoset choroid/RPE during visually guided ocular growth. The identification of novel candidate genes in choroid/RPE of marmoset eyes actively accelerating or decelerating their rates of ocular elongation may elucidate the choroidal response during the regulation of postnatal ocular growth and may lead to the identification of choroid/RPE signaling molecules that participate in scleral remodeling.
Shelton
Lilian
L
Department of Cell Biology, University of Oklahoma Health Science Center, 940 Stanton L. Young Blvd., Oklahoma City, OK 73104, USA.
Troilo
David
D
Lerner
Megan R
MR
Gusev
Yuriy
Y
Brackett
Daniel J
DJ
Rada
Jody Summers
JS
D000818:Animals; D002144:Callithrix / Q000235:genetics*; D002448:Cell Adhesion / Q000187:drug effects; D002478:Cells, Cultured; D002829:Choroid / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism; D004586:Electrophoresis; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007908:Lens, Crystalline / Q000187:drug effects / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010857:Pigment Epithelium of Eye / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism; D012313:RNA / Q000302:isolation & purification; D012029:Refraction, Ocular / Q000187:drug effects / Q000235:genetics*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012590:Sclera / Q000166:cytology; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D015348:Vision, Binocular / Q000187:drug effects; D014822:Vitreous Body / Q000187:drug effects / Q000378:metabolism
D005136:Eye Proteins; D016212:Transforming Growth Factor beta; D012313:RNA