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Run-on gene transcription in human neocortical nuclei

Inhibition by nanomolar aluminum and implications for neurodegenerative disease

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Abstract

The incorporation of [α-32P]-uridine triphosphate into DNA transcription products was examined in short post-mortem interval (PMI) human brain neocortical nuclei (n, 22; PMI, 0.5–24 h) using run-on gene transcription. Reverse Northern dot-blot hybridization of newly synthesized RNA against either total cDNA or Alu repetitive DNA indicated that human brain neocortical nuclei of up to 4-h PMI were efficient in incorporating radiolabel into new transcription products, after which there was a graded decline in de novo RNA biosynthetic capacity. To test the effects of 0–3000 nM concentrations of ambient aluminum on RNA polymerase I (RNAP I) and RNA polymerase II (RNAP II) transcription, dot blots containing 0.5, 1.0, 2.0, and 5.0 µg of DNA for (1) the human-specific Alu repetitive element (2) the neurofilament light (NFL) chain, and (3) glial fibrillary acidic protein (GFAP) were Northern hybridized against newly synthesized radiolabeled total RNA. These DNAs represent heterogeneous nuclear RNA (hnRNA), neuronal-, and glial-specific markers, respectively. We report here a dose-dependent repression in the biosynthetic capabilities of brain RNAP II in the range of 50–100 nM aluminum, deficits similar to those previously described using a rabbit neocortical nuclei transcription system and at concentrations that have been reported in Alzheimer’s disease (AD) euchromatin. Transcription from RNAP II and the neuron-specific NFL gene in the presence of aluminum was found to be particularly affected. These findings support the hypothesis that brain gene transcription in the presence of trace amounts of ambient aluminum impairs mammalian brain DNA to adequately read out genetic information.

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Authors and Affiliations

  1. Neuroscience Center and Department of Ophthalmology, Louisiana State University Medical Center, 2020 Gravier Street, Suite D, 70112, New Orleans, LA

    Walter J. Lukiw & Nicolas G. Bazan

  2. Brain Bank at the LSU Medical Center, Neuroscience Center of Excellence, Louisiana State University School of Medicine, 70112, New Orleans, LA

    Hector J. LeBlanc & Larry A. Carver

  3. Centre for Research in Neurodegenerative Disease, University of Toronto, Tanz Neuroscience Building, M5S 1A8, Toronto, Ontario, Canada

    Donald R. C. McLachlan

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  1. Walter J. Lukiw
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Lukiw, W.J., LeBlanc, H.J., Carver, L.A. et al. Run-on gene transcription in human neocortical nuclei. J Mol Neurosci 11, 67–78 (1998). https://doi.org/10.1385/JMN:11:1:67

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