The zebrafish BMP4 gene: sequence analysis and expression pattern during embryonic development.

We have isolated zebrafish BMP4 gene from a zebrafish genomic DNA library. The size of the isolated BMP4 gene was approximately 14.9 kb. The isolated gene contained two exons which formed the complete coding region together with part of the 3'-noncoding region. The deduced BMP4 protein sequence contained 400 amino acids. Sequence comparison showed that it shared 73% amino acid sequence identity with that of human and mouse BMP4. An intron with a size of 8,963 bp was present between two coding exons. Danio retroposon A (DANA)-like retroposon was located in the intron. It contained four conserved boxes and was flanked by a pair of direct repeats of 9 nucleotide sequence (GTTTTAATA). During embryonic development of the zebrafish, a 3.8-kb BMP4 mRNA was detected from gastrula stage up to a month-old hatching larvae via Northern blot analysis. In addition, the use of reverse transcription polymerase chain reaction further demonstrated the presence of BMP4 mRNA in both the early developmental stages (i.e., cleavage and blastula) and in adult fish. Developmental expression of BMP4 protein was also analyzed. Trace amounts of an 18-kD protein were detected at pharyngula stage, while the production increased from hatching larvae to adult fish. In adult fish, the expression of BMP4 mRNA was observed in brain, heart, digestive tracts, testes, and jaw. The results suggest that the zebrafish BMP4 gene may play important roles during zebrafish development.