Figure 3.
Morphologic analyses in the CNS of treated Glb1-/- mice. (A) Cresyl violet-stained tissue sections of the thalamus, cerebellum, and brain stem from a treated Glb1-/- mouse 3 months after transplantation (Glb1-/- BMT) and from age-matched Glb1+/+ and Glb1-/- mice that underwent mock transplantation (β-gal-/- BMTGFP) revealed restoration of tissue morphology in the treated mouse compared with the extensive vacuolation present in the mice that did not undergo transplantation (Glb1-/-) and the Glb1-/- mice that underwent mock transplantation. (B) Immunolabeling of brain stem, cerebellum, and thalamus revealed the presence of numerous β-gal+ cells in Glb1-/- mice 3 months after BMT. The clear punctate staining demonstrated internalization of the corrective enzyme. Immunolabeling with anti-GFP antibody was done in the same tissues to identify cells of hematopoietic origin. (C) Numerous GFP+ cells in various brain regions showed a ramified microglial morphology. Size bars correspond to 25 μm. Images were visualized using an Olympus BX50 microscope equipped with a 40×/0.65 Plan Apochromatic objective lens (Olympus, Melville, NY) and a Three-Shot 11.3 camera (Diagnostic Instruments, Sterling Heights, MI). Images were acquired with Spot Advanced 4.1.1 software (Diagnostic Instruments) and processed with Adobe Photoshop 8.0 software (Adobe Systems, San Jose, CA).