Human nuclear respiratory factor 2 alpha subunit cDNA: Isolation, subcloning, sequencing, and in situ hybridization of transcripts in normal and monocularly deprived macaque visual system

Nuclear respiratory factor 2 (NRF-2) has been shown to contribute to the tr anscriptional regulation of a number of subunits of respiratory chain enzym es, including cytochrome c oxidase (CO). Our recent study demonstrated a pa rallel distribution of the or subunit proteins of NRF-2 (NRF-2 alpha) with CO in the monkey striate cortex, and that it can be regulated by neuronal a ctivity. To determine whether this regulation is at the transcriptional lev el, the present study examined the expression of NRF-2 alpha mRNA in normal and monocularly deprived adult monkeys. A partial NRF-2 alpha cDNA was iso lated from a human brain cDNA library. Sequence analysis revealed that it s hared 99% identity with the published sequence from human HeLa cells. Ribop robes of NRF-2 alpha was generated and labeled with digoxigenin-11-UTP for in situ hybridization. The expression pattern of NRF-2 alpha mRNA in the no rmal striate cortex paralleled that of CO activity. It was highly expressed in layers IVC and VI, which contained high levels of CO, and more densely expressed in puffs of layers II and III than in interpuffs. In monkeys mono cularly treated with tetrodotoxin for 1 day to 2 weeks, both NRF-2 alpha ex pression and CO activity were reduced in deprived ocular dominance columns of the visual cortex and in deprived layers of the lateral geniculate nucle us. These data indicate that, in the normal and visually deprived adult mon keys, NRF-2 alpha is regulated by neuronal activity at the transcriptional level. J. Comp. Neurol. 417: 221-232, 2000. (C) 2000 Wiley-Liss, Inc.