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. 2008 Feb;172(2):454–469. doi: 10.2353/ajpath.2008.070876

Figure 5.

Figure 5

Induction of autophagy in the hippocampus of neonatal mutant mouse brains after H/I injury. A–D: Triple staining of LC3, TUNEL (TU), and DAPI in the CA1 pyramidal layer of ipsilateral (A, C) and contralateral (B, D) hippocampi deficient in caspase-3 (Casp3−/−) (A, B) and CAD (CAD−/−) (C, D), respectively, 8 hours after H/I injury. Intense granular staining for LC3 is indicated by arrowheads, and granular staining of LC3 in boxed areas are shown in insets (A, C). E–G: Electron micrographs of pyramidal neurons of the hippocampus in mice deficient in caspase-3 (E, F) and CAD (CAD−/−) (G) obtained 3 hours after H/I injury. E and F: Pyramidal neuron containing vacuolar structures in the perikaryal region and showing numerous patches of chromatin clumping in the nucleus (E). Nascent autophagosomes that contained part of the cytoplasm and wrapped by endoplasmic reticulum-like tubular structures were abundantly observed in the boxed area of E (arrowheads in F). G: Part of the perikaryal area of a pyramidal neuron of the hippocampus in a CAD-deficient mouse showing a nascent autophagosome wrapped by an endoplasmic reticulum-like tubule (arrowhead). Scale bars: 10 μm (A–D); 3 μm (E); 1 μm (F and G).