Effect of intracellular photosensitized singlet oxygen production on the electrophysiological properties of cultured rat hippocampal neurons

Photochemical & Photobiological Sciences Pub Date: 2010-10-08 DOI: 10.1039/C0PP00131G

Abstract

Whole-cell patch-clamp recordings from single cultured mammalian neurons have been used to provide insight into early membrane-dependent events that result upon the intracellular photosensitized production of singlet molecular oxygen, O2(a1Δg). The singlet oxygen sensitizers used, pyropheophorbide a (PPa) and protoporphyrin IX (PpIX), locate mainly in cell membranes and mitochondria, respectively. Irradiation of these sensitizers altered both passive and dynamic electrophysiological properties of the neurons in a dose-dependent manner, though the response threshold was much lower with PPa than with PpIX. In particular, notable decreases were observed in the rising and falling rates of action potentials and, at higher light fluences, plateau potentials consistent with activation of Ca2+ channels also developed. The data suggest that singlet oxygen production specifically influences Na+, K+ and Ca2+ ionophores in the cell membrane. Upon terminating sensitizer irradiation, responses evoked by PPa stabilized immediately whereas those evoked by PpIX continued to develop. These data are consistent with a spatially-resolved sphere of intracellular singlet oxygen activity. While the response to PPa irradiation appears to be membrane specific, the response to PpIX irradiation appears to be systemic and possibly part of a cascade of apoptotic events. These results should contribute to a better understanding of membrane-dependent events pertinent to cell death mediated by singlet oxygen.

Graphical abstract: Effect of intracellular photosensitized singlet oxygen production on the electrophysiological properties of cultured rat hippocampal neurons
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