Ultrastructural study of depolarization-induced translocation of NRF-2 transcription factor in cultured rat visual cortical neurons

Shou Jing Yang, Huan Ling Liang, Gang Ning, Margaret T.T. Wong-Riley

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Nuclear respiratory factor (NRF)-2 or GA-binding protein is a potential transcriptional, bigenomic coordinator of mitochondrial and nuclear-encoded subunits of cytochrome oxidase genes. It is composed of an a subunit that binds DNA and a β subunit that has the transactivating domain. Previously, we found that the level of NRF-2 paralleled that of cytochrome oxidase under normal and functionally altered states. The goal of our present study was to increase the resolution to the ultrastructural level and to quantify changes before and after depolarizing stimulation. We used a pre-embedding immunogold-silver method for the two subunits of NRF-2 in cultured rat visual cortical neurons. NRF-2α and β were normally located in both the nucleus and the cytoplasm. In the nucleus, both subunits were associated primarily with euchromatin rather than heterochromatin, consistent with active involvement in transcription. In the cytoplasm, they were associated mainly with free ribosomes and occasionally with the Golgi apparatus and the outer membrane of the nuclear envelope. Labelling was not found in the mitochondria, confirming the specificity of the antibodies. Neuronal depolarization by KCl for 5 h induced a six- to seven-fold increase in the nuclear-to-cytoplasmic ratio of both subunits (P < 0.001) without increases in total labelling densities. These results strongly indicate that both NRF-2α and NRF-2β respond to increased neuronal activity by translocating from the cytoplasm to the nucleus, where they engage in transcriptional activation of target genes. Our results also indicate that the cytoplasmic to nuclear movement of transcription factors is a dynamic process induced by neuronal activity.

Original languageEnglish (US)
Pages (from-to)1153-1162
Number of pages10
JournalEuropean Journal of Neuroscience
Volume19
Issue number5
DOIs
StatePublished - Mar 1 2004

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GA-Binding Protein Transcription Factor
Transcription Factors
Neurons
Cytoplasm
Electron Transport Complex IV
Euchromatin
Antibody Specificity
Heterochromatin
Nuclear Envelope
Golgi Apparatus
Ribosomes
Silver
Transcriptional Activation
Genes
Mitochondria
Membranes

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)

Cite this

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abstract = "Nuclear respiratory factor (NRF)-2 or GA-binding protein is a potential transcriptional, bigenomic coordinator of mitochondrial and nuclear-encoded subunits of cytochrome oxidase genes. It is composed of an a subunit that binds DNA and a β subunit that has the transactivating domain. Previously, we found that the level of NRF-2 paralleled that of cytochrome oxidase under normal and functionally altered states. The goal of our present study was to increase the resolution to the ultrastructural level and to quantify changes before and after depolarizing stimulation. We used a pre-embedding immunogold-silver method for the two subunits of NRF-2 in cultured rat visual cortical neurons. NRF-2α and β were normally located in both the nucleus and the cytoplasm. In the nucleus, both subunits were associated primarily with euchromatin rather than heterochromatin, consistent with active involvement in transcription. In the cytoplasm, they were associated mainly with free ribosomes and occasionally with the Golgi apparatus and the outer membrane of the nuclear envelope. Labelling was not found in the mitochondria, confirming the specificity of the antibodies. Neuronal depolarization by KCl for 5 h induced a six- to seven-fold increase in the nuclear-to-cytoplasmic ratio of both subunits (P < 0.001) without increases in total labelling densities. These results strongly indicate that both NRF-2α and NRF-2β respond to increased neuronal activity by translocating from the cytoplasm to the nucleus, where they engage in transcriptional activation of target genes. Our results also indicate that the cytoplasmic to nuclear movement of transcription factors is a dynamic process induced by neuronal activity.",
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Ultrastructural study of depolarization-induced translocation of NRF-2 transcription factor in cultured rat visual cortical neurons. / Yang, Shou Jing; Liang, Huan Ling; Ning, Gang; Wong-Riley, Margaret T.T.

In: European Journal of Neuroscience, Vol. 19, No. 5, 01.03.2004, p. 1153-1162.

Research output: Contribution to journalArticle

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AU - Yang, Shou Jing

AU - Liang, Huan Ling

AU - Ning, Gang

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