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 transcriptional regulation of a number of subunits of respiratory chain enzymes, including cytochrome c oxidase (CO). Our recent study demonstrated a parallel distribution of the alpha subunit proteins of NRF-2 (NRF-2 alpha) with CO in the monkey striate cortex, and that it can be regulated by neuronal activity. To determine whether this regulation is at the transcriptional level, 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 isolated from a human brain cDNA library. Sequence analysis revealed that it shared 99% identity with the published sequence from human HeLa cells. Riboprobes 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 normal 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 monocularly treated with tetrodotoxin for 1 day to 2 weeks, both NRF-2 alpha expression and CO activity were reduced in deprived ocular dominance columns of the visual cortex and in deprived layers of the lateral geniculate nucleus. These data indicate that, in the normal and visually deprived adult monkeys, NRF-2 alpha is regulated by neuronal activity at the transcriptional level.
Guo
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A
Department of Cell Biology, Neurobiology, and Anatomy, Medical College of Wisconsin, Milwaukee 53226, USA.
Nie
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F
Wong-Riley
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M
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification*; D004268:DNA-Binding Proteins / Q000235:genetics*; D003720:Densitometry; D003576:Electron Transport Complex IV / Q000378:metabolism; D051268:GA-Binding Protein Transcription Factor; D006801:Humans; D017403:In Situ Hybridization; D008252:Macaca fascicularis / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012016:Reference Values; D012683:Sensory Deprivation / Q000502:physiology*; D013779:Tetrodotoxin / Q000494:pharmacology; D014018:Tissue Distribution; D014157:Transcription Factors / Q000235:genetics*; D015349:Vision, Monocular / Q000502:physiology*; D014793:Visual Cortex / Q000378:metabolism
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D051268:GA-Binding Protein Transcription Factor; D012333:RNA, Messenger; D014157:Transcription Factors; D013779:Tetrodotoxin; D003576:Electron Transport Complex IV