Structure and developmental expression of the mouse RGR opsin gene.

PURPOSE The aim of this study is to isolate and characterize cDNA clones and the genes that encode mouse RPE retinal G protein-coupled receptor (RGR) and to analyze expression of the RGR gene in the developing mouse retina. The conserved amino acid sequences of RGR from various mammals can be compared to the amino acid sequence motif of G protein-coupled receptors. METHODS Mouse RGR cDNA and gene clones were isolated from a retina cDNA library and 129SV genomic DNA library, respectively. The expression of RGR in the developing C57BL/6J mouse retina was analyzed by immunohistochemical staining with a polyclonal antipeptide antibody. RESULTS The deduced amino acid sequence of mouse RGR is 78% and 81% identical to that of bovine and human RGR, respectively. The mouse RGR gene is split into seven exons and extends about 11 kb. Two predominant mRNA transcripts, 1.9 and 1.7 kb in length, and a third, relatively faint, 5.5-kb transcript were detected in mouse eye by hybridization to a RGR cDNA probe. Frozen sections of C57BL/6J mouse retina at various stages of development were incubated with a mouse RGR antipeptide antibody. RGR immunoreactivity was first seen at postnatal day 2 (P2) in centrally located RPE cells. From day P6 to P12, there was an increase in the number and intensity of immunoreactive RPE cells in the central and mid-peripheral regions of the retina, while the most peripheral RPE cells were still negative. By day P16, the length of the RPE monolayer was immunoreactive, and staining of the central RPE cells was markedly more intense than at younger ages. CONCLUSIONS Mouse and human RGR are highly conserved. A gradient of RGR expression in RPE extends from the central to the peripheral retina during development. In reference to the appearance of melanin-positive differentiated RPE cells, the induction of RGR expression is a relatively late event in the maturation of the retina.