Abnormal retinal pigment epithelium melanogenesis as a major determinant for radiation-induced congenital eye defects.

• X-ray exposure at E7.5 decreases gene and protein expression of Pax6 and Lhx2 at E9. • Irradiation at E7.5 decreases melanogenic gene expression at E11. • Reduced embryonic eye pigmentation at E11 may precede radiation-induced eye defects. • Radiation exposure during neurulation decreases retinal mitosis at E18. Recent studies highlighted a link between ionizing radiation exposure during neurulation and birth defects such as microphthalmos and anophthalmos. Because the mechanisms underlying these defects remain largely unexplored, we irradiated pregnant C57BL/6J mice (1.0 Gy, X-rays) at embryonic day (E)7.5, followed by histological and gene/protein expression analyses at defined days. Irradiation impaired embryonic development at E9 and we observed a delayed pigmentation of the retinal pigment epithelium (RPE) at E11. In addition, a reduced RNA expression and protein abundance of critical eye-development genes (e.g. Pax6 and Lhx2) was observed. Furthermore, a decreased expression of Mitf, Tyr and Tyrp1 supported the radiation-induced perturbation in RPE pigmentation. Finally, via immunostainings for proliferation (Ki67) and mitosis (phosphorylated histone 3), a decreased mitotic index was observed in the E18 retina after exposure at E7.5. Overall, we propose a plausible etiological model for radiation-induced eye-size defects, with RPE melanogenesis as a major determining factor. [ABSTRACT FROM AUTHOR]