Sequences and expression patterns of alkaline phosphatase isozymes in preattachment bovine embryos and the adult bovine

We report the cloning and partial sequences of two novel bovine tissue-specific alkaline phosphatase (AP) isozymes (TSAP2 and TSAP3) from in vitro-produced bovine blastocysts. Using a reverse-transcribed polymerase chain reaction (RT-PCR)-based assay for mRNA expression and in vitro-produced preattachment bovine embryos, TSAP2 mRNA was detected first at the four-cell stage prior to the major burst of embryonic transcription in cattle and TSAP3 at the eight-cell stage with the major burst in transcription. Furthermore, the transcription of TSAP2 and TSAP3 displays a curious "on-off" pattern during early cleavages between 40 and 120 hr after insemination. Activity of bovine AP, measured by an azo-dye coupling technique, indicates that at least one AP isozyme is functional in oocytes and embryos throughout bovine preattachment development. However, maternal and embryonic-derived AP activity may have different cell-surface distributions. This novel expression pattern of the bovine AP isozymes could provide a useful tool for identifying and clarifying the events controlling transcription and gene expression during early embryo development.