Figure 5:

Bovine testicular hyaluronidase and rPH20 block OPC maturation in a model of perinatal white matter injury. (A) Montage of low magnification images of a brain slice including the corpus callosum (outlined) stained with DAPI (blue), MBP (green), and olig2 (red), demonstrating the area where slices were analyzed for changes in OPC maturation. (B) A representative slice treated with vehicle and stained for MBP (white). Inset in B: high magnification image from the same slice, showing the morphology of a maturing oligodendrocyte. (C) A representative slice treated with 20 U/ml BTH and stained for MBP (white). Scale bars = 500 μm. Dotted lines outline the corpus callosum. (D) Quantification of MBP labeling in slices treated with vehicle or BTH. (E) Immunostaining of O4 (green) and O1 (red) from a representative slice treated with vehicle. (F) Immunostaining for O4 and O1 from a representative slice treated with 20 U/ml BTH. Scale bars = 20 μm. (G-I) Quantification of immunostaining in slice cultures for O4 (G), O1 (H) and CC1 (I). (J) Quantification of immunostaining from slices treated with vehicle or 20 U/ml of rPH20. For the MBP experiments, sham = 6 animals; hyaluronidase = 5 animals (both from 3 independent culture days). We analyzed ~3 slices from each animal (some removed because of poor viability), with a total of 10 slices in the PBS control and 12 slices in the hyaluronidase group. For the O4/O1 experiments, sham = 4 animals; hyaluronidase = 5 animals (both from 3 independent culture days). We used a total of 15 slices in the control group, and 15 slices in the hyaluronidase group. Comparisons were analyzed using a Student’s t-test. *p<0.001.