The expression of the RAS and podocin proteins was also altered. Furthermore, not only the F1 generation but also the F2 generation was affected by the gestational vitamin D deficiency of the F0 generation mice. Vitamin D is normally stored in the liver and adipose tissue, and after six weeks on a restricted diet, the vitamin D levels were extremely reduced without alteration of calcium or phosphate levels in the rodents. Previous studies in PD325901 rodents have shown that the restriction of vitamin D for a period of six weeks prior to mating assures vitamin D deficiency during pregnancy. Vitamin D deficiency during the third trimester of pregnancy in humans has been associated with cardiovascular disease, diabetes and obesity in offspring, which could be linked to the potential role of vitamin D in the regulation of adiposity. Maternal nutrient restriction causes adiposity in offspring, and recent data suggest that maternal vitamin D deficiency affects adiposity, although the mechanisms are not yet fully understood. In the present study, adiposity was not the focus. However, there was a significant increase in BM in both generations of VitDoffspring at six months of age, but the BM was higher only in the F2-VitD- group at weaning. In three different moments of postnatal life, BM was not affected by the maternal diet restricted in vitamin D. At weaning and at six months of age, the VitD- offspring were heavier than the SC offspring in both the F1 and F2 generations. However, at three months of age, the VitD- groups in both the F1 and F2 generations showed a tendency toward increased BM, although the difference was not statistically significant. The small sample size of the groups could explain these results. In rats, it has been observed that maternal vitamin D deficiency leads to a significant increase in the number of glomeruli, which is a unique and important effect of vitamin D restriction during fetal development. To understand this result, it should be emphasized that nephrogenesis in rodents, unlike in humans, continues after delivery for the first 10 days of postnatal life. This period is roughly comparable to the third gestational trimester of humans. Experimental models have demonstrated that blocking the RAS during development causes changes in the glomerular vasculature that affects the normal maturity of the kidney, although the RAS cascade during kidney development is not fully understood. Research suggests that vitamin D is important for differentiation and maturation during cell proliferation and that it downregulates renin gene expression in the kidneys. However, in the present study, renin expression was significantly higher at six months of age in both the F1 and F2 generations, and AT1r expression was higher in F1-VitD- offspring than in F1-SC offspring. Kidney organogenesis is begun and maintained as a result of a series of regulatory molecules. WT1 is expressed in the metanephric blastema and probably plays a key role in podocyte differentiation. In the present study, no significant reduction in the expression of WT1 in the kidneys at birth was observed in either the F1 or F2 generation. Although the VitDoffspring in the present study did not show a reduction in WT1 expression, maternal vitamin D deficiency did alter the ratio of immature-to-mature glomeruli at birth by extending the period of glomerular maturation. Maternal vitamin D deficiency may also have contributed to an increased number of glomeruli, thus producing smaller glomeruli than in animals fed SC. Indeed, the smaller glomeruli could explain the high BP observed among F1-VitD- offspring and the strong tendency toward high BP observed in the F2 generation. Previous research has documented a negative correlation between the number of glomeruli and BP levels.