Effects of mycophenolate mofetil (MMF) treatment on glial apoptosis and proliferation.
(A) High-magnification confocal laser scanning microscopy images, double-labeled with Ki-67 (proliferating cells, red) in combination with either isolectin (I)B4 (microglial cells, green) or glial fibrillary acidic protein (astrocytes, green). Effects of MMF on (B) microglial and (C) astroglial proliferation indices at 36 hours or 48 hours post-lesion compared with lesioned organotypic hippocampal slice cultures (OHSCs) that received additional MMF treatment within 12 to 36 hours or 24 to 48 hours, respectively (*P < 0.05). (D) Confocal laser scanning microscopy images at high magnification, double-labeled with cleaved caspase-3 (apoptotic cells, red) with either IB4 (microglial cells, green) or GFAP (astrocytes, green). (E,F) Effects of MMF on microglial and astroglial apoptosis indices at 36 or 48 hours post-lesion compared with lesioned OHSCs that received additional MMF treatment within 12–36 hours or 24 to 48 hours, respectively (*P < 0.05). MMF treatment within the crucial neuroprotective 12–36-hour time window potently attenuated microglial and astrocytic proliferation, but did not affect rates of glial apoptosis. Application of MMF within 24 to 48 hours post-lesion significantly reduced microglial proliferation but did not affect either astrocytic proliferation or glial apoptosis. Scale bars = 25 μm