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Fig. 4 | Journal of Neuroinflammation

Fig. 4

From: Escalating morphine dosing in HIV-1 Tat transgenic mice with sustained Tat exposure reveals an allostatic shift in neuroinflammatory regulation accompanied by increased neuroprotective non-endocannabinoid lipid signaling molecules and amino acids

Fig. 4

Morphine treatment increased the number of microglia (Iba-1+) and morphine and Tat exposure increased activated microglia (3-NT+ Iba-1+) in the striatum and spinal cord of Tat transgenic mice. a Quantification of Iba-1+ cell counts in 5 different CNS regions, including the frontal cortex, hippocampus, striatum, cerebellum, and spinal cord were examined for Iba-1+ cells. Morphine upregulated Iba-1+ cell counts in the striatum and spinal cord, specifically for the morphine-treated, Tat(+) mice. Tat expression showed no significant effect. CNS sections from the striatum and spinal cord stained for Iba-1+ cells (green) taken at × 20 with a higher magnification images taken at × 63. Scale bars = 50 μm. b Immunofluorescent images were taken in the spinal cord at × 63 and show double-labeling for the microglial marker Iba-1 (green) and 3-nitrotyrosine (3-NT; red), a reactive nitrosyl product detected in activated microglia, with containing Hoechst-stained nuclei. The 3-NT+ Iba1+ double-labeled microglia from a saline-treated Tat(−) mouse is ramified and shows relative low level of 3-NT expression. In contrast the 3-NT+ Iba1+ double-labeled microglia from the morphine-treated Tat(+) mouse shows decreased processes and the level of 3-NT is noticeably higher (see Table 2 for quantification via Sholl analysis). Scale bars = 20 μm. Percent 3-NT+ Iba-1+ cell counts based on total number of Iba-1 cells was quantified in the striatum and spinal cord. Morphine and Tat expression increased 3-NT+ Iba-1+ cell counts (%) in the striatum and spinal cord, specifically for morphine-treated Tat(+) mice. c Example of a single microglial cell with concentric Sholl radii (purple circles) superimposed on the image. Scale bar = 20 μm. All data are expressed as mean ± SEM. Statistical significance was assessed by ANOVA followed by Tukey’s post hoc test; *p < 0.05 main effect of drug; #p < 0.05 main effect of genotype; §p < 0.05 vs. morphine-exposed Tat(+) mice. Morph short-term (8-day) morphine injections. n = 3–4 mice per group/4 sections each

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