Fluorescence staining of F-actin in cultured astrocytes isolated from wild-type, MMP-2 null, and MMP-9 null mice using Texas Red-conjugated phalloidin.

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Fluorescence staining of F-actin in cultured astrocytes isolated from wild-type, MMP-2 null, and MMP-9 null mice using Texas Red-conjugated phalloidin. Fluorescence staining of F-actin in cultured astrocytes isolated from wild-type, MMP-2 null, and MMP-9 null mice using Texas Red-conjugated phalloidin. Shown here are astrocytes without cytoskeleton inhibitor (i), treated with cytochalasin D (ii), and treated with Rac1 inhibitor (iii). Before reaching a confluent monolayer (A–C), astrocytes without cytoskeleton inhibitor exhibit comparable morphology and F-actin distribution in the wild-type and MMP-2 null groups, whereas MMP-9 null astrocytes lose their stellar profile and show disintegrated F-actin distribution (C-i). After a scratch wound is made on the confluent monolayer (D–F), wild-type and MMP-2 null astrocytes, without cytoskeleton inhibitor, at the scratch border extend F-actin-containing membrane protrusions toward the denuded area (on the left). These membrane protrusions are not observed in MMP-9 null astrocytes (F-i). Regardless of having a scratch wound or not, treatments with cytochalasin D (A–F-ii) or Rac1 inhibitor (A–F-iii) cause abnormal morphology and F-actin distribution in astrocytes of all 3 genotypes. No scratch-induced membrane protrusion is found with these treatments. Scale bars: -i, 25 μm; -ii, -iii, 100 μm. Jung-Yu C. Hsu et al. J. Neurosci. 2008;28:13467-13477 ©2008 by Society for Neuroscience