Abstract
Abstract: :
Purpose: Retinal pigment epithelium (RPE) is essential for retina integrity and vision. The CD36 scavenger receptor is involved in photoreceptor outer segment phagocytosis, angiogenesis regulation and immunity and may play an important role in RPE and retina homeostasis. We took advantage at the CD36 knockout mouse to identify and characterize downstream pathways activated or regulated by CD36, using microarray. Methods: Eyes from 1 month–old wild–type and CD36 knockout mice were removed immediately after sacrificing the animals. The neural retina was dissected and, after enzymatic treatments and washes, RPE patches of cells were collected. RNAs from control and CD36 knockout RPE were extracted and cDNA synthetized. Biotinylated cRNA were generated and used as probes for microarrays. Results: We used the Mouse 430A chips from Affymetrix that contains over 22,000 sequences. Preliminary data show that a small number of genes are induced or repressed by at least 2–fold in the RPE lacking CD36, while most genes show no significant change. Genes of interest will be further studied by semi–quantitative PCR, immunohistochemistry and biochemistry. Conclusions: Microarray profiling allowed us to identify differentially expressed genes in CD36 knockout mouse RPE. Since defects in phagocytosis, angiogenesis and immunity are common features that lead to retinal degeneration, several of these genes may be essential for the retina and could further be studied as candidates for retinal diseases.
Keywords: retinal pigment epithelium • gene microarray • gene/expression