FIGURE 5.

myo-Inositol supplementation during pregnancy rescues the lethality and craniofacial defects of Lys-95 homozygotes. A, schematic of the protocol for myo-inositol supplementation. B, Alcian blue and alizarin red staining of rib cages at E18.5. Typical pictures show wild-type (Wild) and homozygote (Homo) mice. Note that malformation of the jaw or rib cage is not visible in homozygotes. C, there was no gross difference between adult wild-type and homozygote mice (left panel). Abnormal lower jaw morphology was rescued by dietary supplementation with inositol during pregnancy (right panels). Top right, ventral view; bottom right, lateral view. D, there were no gross structural abnormalities in the brains of homozygotes stained with hematoxylin and eosin. Cx, cerebral cortex; Hpc, hippocampus; Cb, cerebellum. E, expression level of the Impa1 protein in homozygotes was comparable with that of wild-type controls. Lysates (40 μg of protein) prepared from K95/K95 and control mice were analyzed by Western blotting (WB) with anti-Impa1, anti-Impa2, or anti-Gapdh antibodies. FC, frontal cortex; CB, cerebellum; KN, kidney, and TT, testis. F, Kaplan-Meier survival analysis of mice rescued with myo-inositol. Homozygotes (Homo) showed normal morphology, but their life spans were shorter than those of wild-type (Wild) controls. Note that after weaning, mice drank water without myo-inositol.