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. 2018 Feb 1;12:22. doi: 10.3389/fncel.2018.00022

FIGURE 7.

FIGURE 7

Protection of CA1 neurons from OGD-induced death by SPARC is mediated through Phx-433 sensitive GluA1-containing AMPARs. Organotypic hippocampal slices were exposed to oxygen and glucose deprivation (OGD) for 45 min followed by normoxic reoxgenation for 72 h. Percentage of apoptotic cells was assessed using propidium iodide (PI) labeling of apoptotic nuclei as shown in (A). (A,B) Organotypic hippocampal slices were either pre-treated with SPARC (0.5 μg/ml for 48 h prior and following OGD in the reoxygenation period: chSPARC) before OGD or only acutely post OGD during the reoxygenation period (0.5 μg/ml: acSPARC). SPARC had a significant protective effect on CA1 neurons when slices were pretreated prior to OGD. (C,D) The protective effect of SPARC was inhibited by the AMPAR blocker Phx-433 (5 μM) (A,D). Phx-433 alone had no effect on survival of CA1 neurons either in control or OGD conditions (C). [ANOVA with post hoc Holm–Sidak test (n = 5), ∗p < 0.05]. Scale = 50 μm.