Rescue of retinal morphology and function in a humanized mouse at the mouse retinol-binding protein locus.

Retinol-binding protein RBP4 is the specific carrier for retinol in the blood. We previously produced a Rbp4-deficient (Rbp4-/- ) mouse that showed electroretinogram (ERG) abnormalities, accompanied by histological and electron-microscopic changes such as fewer synapses in the inner plexiform layer in the central retina. To address whether human RBP4 gene expression can rescue the phenotypes observed in Rbp4-/- mice, we produced a humanized (Rbp4hRBP4orf/ hRBP4orf ) mouse with a human RBP4 open reading frame in the mouse Rbp4 locus using a Cre-mutant lox recombination system. In Rbp4hRBP4orf/hRBP4orf mice, the tissue-specific expression pattern of hRBP4orf was roughly the same as that of mouse Rbp4. ERG and morphological abnormalities observed in Rbp4-/- mice were rescued in Rbp4hRBP4orf/hRBP4orf mice as early as 7 weeks of age. The temporal expression pattern of hRBP4orf in the liver of Rbp4hRBP4orf/hRBP4orf mice was similar to that of mouse Rbp4 in Rbp4+/+ mice. In contrast, hRBP4orf expression levels in eyes were significantly lower at 6 and 12 weeks of age compared with mouse Rbp4 but were restored to the control levels at 24 weeks. The serum hRBP4 levels in Rbp4hRBP4orf/hRBP4orf mice were approximately 30% of those in Rbp4+/+ at all ages examined. In accordance with this finding, the plasma retinol levels remained low in Rbp4hRBP4orf/hRBP4orf mice. Retinol accumulation in the liver occurred in control and Rbp4hRBP4orf/hRBP4orf mice but was higher in Rbp4hRBP4orf/hRBP4orf mice at 30 weeks of age. Mouse transthyretin expression was not altered in Rbp4-/- or Rbp4hRBP4orf/hRBP4orf mice. Taken together, 30% of the serum RBP4 level was sufficient to correct the abnormal phenotypes observed in Rbp4-/- mice.