Figure 4.

Confocal microscopic images and quantification of apoptotic bodies in the brain tissue. Morphine treatment in the presence of TAT and S. pneumoniae (S.p.) markedly increases CNS apoptosis. A, To determine apoptosis, animals (6 per group) were treated as described in Figure 1 and killed at 72 h. Brains were removed and snap frozen in liquid nitrogen. Cryostat sections (5 μm) were used to evaluate apoptosis using TUNEL staining (Intergen) according to the manufacturer's instruction. Anatomical location from where sections were taken is indicated (top). DAPI staining shows the nuclei of cells. Scale bars, 10 μm. B, The number of apoptotic cells were also determined using metamorph software and expressed as percentage of total cells. C, The graph represents quantification of apoptotic cells in TATtg mouse brain treated with morphine/placebo pellet and S. pneumoniae. The data are presented as mean ± SD of three independent experiments. *p < 0.01.