Mouse cytochrome b561: cDNA cloning and expression in rat brain, mouse embryos, and human glioma cell lines.

A cDNA encoding cytochrome b561 has been isolated from a mouse brain cDNA library, which predicts a protein of 250 amino acids with a deduced Mr of 27,770. Northern blot analysis of different mouse and rat tissues revealed one major mRNA of 3300 bp, which is abundantly distributed in a number of neuroendocrine tissues. In addition, cytochrome mRNA levels in rat brain sections showed the highest distribution of cytochrome b561 in the hypothalamus, hippocampus, thalamus, and striatum, with a moderate level in the cerebral cortex, and the lowest levels in the olfactory bulb and cerebellum. Because non-neuronal cells in the central nervous system contained peptidyl alpha-amidating monooxygenase (PAM), to which cytochrome b561 donates its electrons, we used RT-PCR to document the coexpression of cytochrome b561 with PAM and dopamine beta-hydroxylase (DBH) in glioblastoma cells. Cytochrome b561 expression was detectable in the 11-day-old mouse embryo, and the level of its mRNA increased tenfold by 15 and 17 days of gestation.