Analysis of amelogenin mRNA during bovine tooth development

The amelogenins are highly conserved enamel-matrix proteins that are essential for proper mineral formation. Transcriptionally active genes encoding the bovine amelogenin proteins reside on both the X and Y chromosomes. Comparison of relative levels of amelogenin mRNAs at various stages of development indicated that the X-chromosomal amelogenin message is at least six fold more abundant than the Y. Alternative splicing generates at least seven messages, five from the X primary transcript, and two from the Y. The two most abundant X-chromosomal amelogenin messages are approx. 850 and 450 nucleotides long, and nearly 10-fold more 850-nucleotide mRNA can be measured than 450 nucleotide, which has lost most of exon 6 by splicing. The predominant small message encodes leucine-rich amelogenin protein (LRAP), and amounts of LRAP message are relatively constant during development. However, the amelogenin message from which exon 3 has been spliced declines approximately 2.3-fold, when compared to total X chromosomal amelogenin transcripts, suggesting differential regulation of alternative splicing. In addition, a new exon was identified within genomic DNA, which was shown to be expressed by the use of reverse transcriptase-polymerase chain reaction, and the exons were renamed accordingly. This new exon-4 sequence is unusual in that it is not highly conserved between species.