Improvement in neuronal survival after ischemic preconditioning in hippocampal slice cultures

Guang Ping Xu, Kunjan R. Dave, Richard Vivero, Rainald Schmidt-Kastner, Thomas J. Sick, Miguel A. Pérez-Pinzón

Research output: Contribution to journalArticle

69 Scopus citations

Abstract

The main goals of the current study were to assess: (a) whether a sublethal ischemic insult could protect the CA1 subregion of the hippocampus in organotypic slices against a lethal ischemic insult; and (b) whether this protection is long lasting as determined with an accurate immunohistochemical neuronal marker, NeuN. Hippocampal slice cultures were grown for 12-14 days in vitro. Slices were exposed either to oxygen/glucose deprivation (OGD) for 45 min (ischemia), or OGD for 15 min (ischemic preconditioning), 48 h prior to 45 min OGD, or were untreated (sham). Cell death was estimated by propidium iodide fluorescence 1 day after OGD and by NeuN immunohistochemistry 7 days after OGD. Image analysis was employed to measure the relative optical density of the NeuN-signal in all groups. After ischemia, damaged neurons were shrunken or lost and NeuN immunoreactivity was reduced. Relative optical density of NeuN (ROD [NeuN]) was 0.193±0.015 in control (sham) (n=9). In slices that underwent ischemia, ROD [NeuN] declined to 0.108±0.018 (n=5) in CA1 (*P<0.05 ROD [NeuN] in preconditioned slice cultures was 0.190±0.037 (76% higher than the ischemia group). Similar results were found after measuring PI fluorescence. In the CA1 sub-region, PI fluorescence was about 13, 47 and 17% in the sham, ischemic and IPC groups, respectively. We suggest that the immunohistochemical approach validates the dye uptake method used in slice cultures and yields quantitative data specific for neurons. We also conclude that the organotypic hippocampal slice model is useful for studying delayed ischemic preconditioning that is maintained for hours or days after the preconditioning event.

Original languageEnglish (US)
Pages (from-to)153-158
Number of pages6
JournalBrain Research
Volume952
Issue number2
DOIs
StatePublished - Oct 18 2002

Keywords

  • Anoxia
  • Cell culture
  • Ischemia
  • Tolerance

ASJC Scopus subject areas

  • Neuroscience(all)

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