Enamel defects have been reported in rickets and related to disturbed expression of amelogenin in ameloblasts. The present study is devoted to amelogenin, enamelin, ameloblastin, and dentin sialo-phosphoprotein (DSPP) expression in both the epithelium and mesenchyme of vitamin D-deficient rat incisors. Quantitative Northern blotting analysis (relatively to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA steady-state levels) was performed on microdissected cells of rachitic (-D) and control (+D) 56 day-old rats. Steady-state levels of amelogenin and enamelin mRNA were significantly reduced in the -D epithelium versus the +D epithelium ones. In contrast, ameloblastin expression was slightly increased in -D epithelium. In the same samples, DSPP mRNA levels remained unchanged in -D dental mesenchyme. Comparative electron microscopy studies between +D and -D animals showed a dramatic decrease of intraprismatic enamel (amelogenin and enamelin immunoreactive) consistent with our molecular results. In conclusion, tooth formation results from the coordinated expression of several matrix proteins that may be controlled by 1,25-dihydroxyvitamin D-3 [1,25(OH)(2)D-3].