Ogression of Purkinje cell loss in regions with activated Bergmann glia (Fig. 5b-d). Practically all areas with activated Bergmann glia displayed prominent Purkinje cell loss at 20 weeks although transgene expression was inactivated in the period of 12 to 20 weeks of age (Fig. 5b-d). This shows that Purkinje neurons are unable to survive for an extended time period in an atmosphere of activated Bergmann glia.Lattke et al. Molecular Neurodegeneration (2017) 12:Web page eight ofFig. 4 IKK2-CA expression in Bergmann glia disrupts their morphology in correlation using the stage of cerebellar degeneration. a Co-staining for hIKK2 (transgene) and also the astrocyte marker Aldh1l1 at 20 weeks of age in a cerebellum devoid of detectable degeneration (“early deg”) and within a late stage of degeneration (“late deg”) Arrows: Bergmann glia cell bodies. b-d IKK2-CA-expressing (hIKK2 positive) Bergmann glia marked by arrows show nuclear localization of RelA indicating active NF-B signalling (b). Arrowheads: Purkinje cell bodies (hIKK2 negative). c Enlargement of the box in B. d Quantification of nuclear RelA immunofluorescence intensity in cells inside the Purkinje cell layer, like IKK2-CA (hIKK2) expressing Bergmann glia. n = 360 cells of 3 handle and n = 480 of four IKK2-CA animals. Statistical evaluation: 1-way ANOVA with Tukey’s post-test, p sirtuininhibitor 0.001. e Staining for GFAP at 20 weeks of age shows Bergmann glia processes within the molecular layer. Controls and IKK2-CA cerebella without detectable degeneration (“early deg”) show parallel Bergmann glia processes with weak GFAP. Severely degenerated cerebella (“late deg”) show intensely stained, thickened and unorganized processes. f Occasionally, IKK2-CA animals without the need of clear Purkinje cell loss (age 12 weeks) show patches of Bergmann glia with improved GFAP expression and first indicators of disorganisation. Merged images show DAPI co-staining (blue). Scale bars: 20 mConsistent with this model, in animals with continuous transgene expression some areas show Bergmann glia activation, but no or only moderate Purkinje cellloss (Fig.Vitronectin Protein MedChemExpress 5a and c-d), indicating that Bergmann glia in these areas have already been activated only lately, and for that reason the resulting delayed Purkinje cell loss continues to be inLattke et al.Adrenomedullin/ADM Protein medchemexpress Molecular Neurodegeneration (2017) 12:Web page 9 ofFig.PMID:24220671 five (See legend on next page.)Lattke et al. Molecular Neurodegeneration (2017) 12:Web page ten of(See figure on prior web page.) Fig. 5 Nearby IKK2-CA-induced Bergmann glia activation drives subsequent Purkinje cell loss. a, b Variable loss of Purkinje cells (calbindin-positive) in locations with astrogliosis-like Bergmann glia activation (improved GFAP staining) in IKK2-CA animals at 20 weeks. Places devoid of detectable degeneration (“early deg”) showing parallel Bergmann glia processes and degenerated places (“late deg”) are depicted (a). After IKK2-CA repression by doxycycline from 12 weeks of age extreme Purkinje cell loss at 20 weeks is evident in areas with Bergmann glia activation, but not in fields with normal Bergmann glia (b). Inlays: higher magnification of GFAP channel within the molecular layer. DAPI co-staining (blue). Scale bars 50 m (inlays 20 m). c, d Quantitative evaluation in the spatial correlation of Bergmann glia activation (sirtuininhibitor8 location GFAP+ in the molecular layer) and Purkinje cell degeneration (sirtuininhibitor15 Purkinje cells/mm) in individual microscopic fields. Degeneration is virtually exclusively located in fields with activated Bergmann glia (c). Be.
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