4A). Both IGF-1 and HGF ameliorate 6-OHDA-induced Parkinsonism (Clarkson et al. 2001; Koike et al. 2006; Ebert et al. 2008). mRNAs encoding the neuroprotective factors

IGF-1 and HGF increased, and mRNAs for the detrimental proinflammatory cytokines decreased in the presence of the both cytokines. Thus, this result indicates that PH-797804 manufacturer GM-CSF and IL-3 strengthened the neuroprotective nature of cultured microglia. Similar results were obtained in in vivo experiments (Fig. 4B). IGF-1 and HGF-mRNAs increased more significantly in the ventral midbrain of the cytokine group than in the ventral midbrain of the saline group. IL-1β and TNFα mRNAs markedly increased in the Inhibitors,research,lifescience,medical saline group, but the levels in the cytokine groups returned to the sham level. Immunohistochemical staining Inhibitors,research,lifescience,medical using anti-IL-1β and TNFα antibodies showed the positive immunostaining of the proinflammatory cytokines in DArgic neurons (blue arrowheads) and microglia (yellow ones) (Fig. 4C–F). In spite of such functional differences, microglial cells in the SNpc displayed amoeboid morphology in both the 6-OHDA-treated groups. Figure 4 Effects of cytokines on microglial cells in vitro and in vivo. (A) qRT-PCR revealed that cultured microglial cells

have increased mRNAs encoding IGF-1 (Aa) and HGF (Ab), while Inhibitors,research,lifescience,medical there is decreased mRNAs encoding IL-1β (Ac) and TNFα (Ad), … Contact between neurons and glia Detailed morphological Inhibitors,research,lifescience,medical observation using 3D-constructed images taken by CLSM revealed the intimate contacts between neurons and glial cells and the presence of Iba1+/NG2+ cells (Fig. 5). The brain section in Figure 5 was from a cytokine-injected rat that was immunostained with antibodies to Iba1, NG2, and TH. The merged image of Iba1 and NG2 immunoreactivities (Fig. 5D) shows the presence of Iba1+/NG2+ cells, which have been described as a neuroprotective cell type (Kitamura et al. 2010). Activated microglial cells have long been Inhibitors,research,lifescience,medical described to intimately

attach to damaged neurons and remove synaptic inputs. This phenomenon is called “synaptic stripping” and is supposed to be neuroprotective (Cullheim and Thams 2007; Trapp et al. 2007). The presence of synaptic stripping by immunofluorescence would be evident when the green fluorescence representative of Iba1-immunoreactivity is merged CYTH4 with the red fluorescence of TH-immunoreactivity, thus producing yellow color. Indeed, the merged yellow color is evident in the region where microglia and DArgic neurons intimately attach in Fig. 5E. In addition, NG2 glia also appeared to closely attach to DArgic neurons. This is seen when green immunofluorescence of NG2 is merged with the red immunofluorescence of DArgic neurons; the contact regions of NG2 glia and DArgic neurons appear as orange regions (Fig. 5F). The attachment of NG2 glia to DArgic neurons appeared more frequently than that of microglia.