Precocious IRBP Gene Expression During Mouse Development

Purpose: To determine if the time course for the onset of gene and protein expression for interphotoreceptor binding protein (IRBP) precedes that of opsin in the developing mouse retina.

Methods: Relative mRNA levels of the IRBP and opsin genes were determined in prenatal and postnatal retinal RNA with RNase protection analysis (RPA). To determine if IRBP and opsin protein expressions are differentially regulated, dissociated retinal cells from postnatal (P) days 2 and 3 mice, that were injected with BrdU, were then double-labeled with antibodies against BrdU and either opsin or IRBP.

Results: With RPA, IRBP mRNA was detected on embryonic (E) day 11 at the time of cone formation, whereas opsin mRNA was not detected until P0. It took until P3 for opsin expression to reach significant levels, whereas rods already appear during embryonic development. IBRP transcription preceded that of opsin because it rapidly increased from E13 to an early postnatal day. By P20, the expression levels of IRBP and opsin achieved constancy. Double antibody labeling revealed positive staining for both IRBP and BrdU as soon as 2 hours after injection, but it took until 40 hours for double positive staining for opsin and BrdU.

Conclusion: Because only IRBP protein expression was observed before the last mitosis of the photoreceptor precursor cells, IRBP could be essential for retinal development.