Using 31P NMR spectroscopy at 14.1 Tesla to investigate PARP-1 associated energy failure and metabolic rescue in cerebrocortical slices

J Bioenerg Biomembr. 2004 Aug;36(4):415-9. doi: 10.1023/B:JOBB.0000041777.07640.64.

Abstract

PARP-1 activation by H(2)O(2) in an acute preparation of superfused, respiring, neonatal cerebrocortical slices was assessed from PAR-polymer formation detected with immunohistochemistry and Western blotting. (31)P NMR spectroscopy at 14.1 Tesla of perchloric acid slice extracts was used to assess energy failure in a 1-h H(2)O(2) exposure as well as in a subsequent 4-h recovery period where the superfusate had no H(2)O(2) and specifically chosen metabolic substrates. Although more data are needed to fully characterize different bioenergetic responses, a high NMR spectral resolution (PCr full-width at half-max approximately.01 ppm) and narrow widths for most metabolites (<.2 ppm) permitted accurate quantifications of spectrally resolved resonances for ADP, ATP, NAD(+)/NADH, and other high energy phosphates. It appears possible to use brain slices to quantitatively study PARP-related, NAD-associated energy failure, and rescue with TCA metabolites.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Brain / drug effects
  • Brain / metabolism*
  • Cell Hypoxia / drug effects
  • Cell Hypoxia / physiology
  • Energy Metabolism / drug effects
  • Energy Metabolism / physiology
  • Hydrogen Peroxide / administration & dosage
  • Magnetic Resonance Imaging / methods*
  • Magnetics
  • NAD / metabolism*
  • Oxidative Stress / drug effects
  • Oxidative Stress / physiology
  • Phosphorus Isotopes
  • Poly (ADP-Ribose) Polymerase-1
  • Poly(ADP-ribose) Polymerases / metabolism*
  • Rats
  • Rats, Sprague-Dawley
  • Tissue Culture Techniques / methods

Substances

  • Phosphorus Isotopes
  • NAD
  • Hydrogen Peroxide
  • Parp1 protein, mouse
  • Poly (ADP-Ribose) Polymerase-1
  • Poly(ADP-ribose) Polymerases