Molecular cloning from neurulating Ambystoma mexicanum embryos of the cDNA encoding an orphan nuclear receptor (aDOR1) closely related to TR2-11.

We have isolated a cDNA encoding a novel orphan nuclear receptor, aDOR1, closely related to testicular receptor-2 (TR2) orphan receptor family members, from neurulating Ambystoma mexicanum embryos. The cDNA sequence predicts a protein primary sequence of 416 amino acids with a calculated molecular weight of 45.8 kDa. While the DNA-binding domains of aDOR1 and hTR2-11 share 96% identity, considerable divergence is observed at both extremities of the peptides. At the N-terminus, aDOR1 is 66% identical to hTR2-11 and longer by 37 amino acids. At the C-terminus, despite a greater similarity (69%), aDOR1 is much shorter than the hTR2 isoforms and seems to encode a distinct ligand-binding domain. Expression of aDOR1 was studied by the reverse transcription polymerase chain reaction assay (RT-PCR). High mRNA levels were detected during oogenesis, they remained high during the cleavage stage, and decreased at the mid-blastula transition (MBT). Transcripts increased again at the end of gastrulation, reached a peak level during neurulation, and leveled off after closure of the neural tube. In neurulas dissected along the anteroposterior axis, aDOR1 mRNA was enriched at both extremities of the embryo, while no particular distribution was favored along the dorsoventral axis. Retinoic acid (RA) treatments at the beginning of gastrulation did not affect overall mRNA levels in the neurula nor its distribution along both axes. In the adult, expression was predominant in the brain; lower levels (about 15%) were detectable in all germ layer derivatives, except muscle. These results suggest that aDOR1 may be required for the early determination events occurring during the cleavage stages of development, and may be involved in embryogenesis and in brain function.