Page - 23 - in Advances in Neuroimmunology

BrainSci. 2016,6, 15 Figure 2. Assessment of the total number of cellular infiltrates (A) and demyelination (B) in the longitudinal lumbar-cervical spinalcordwhitematter fromEAE-inducedanimalsat20dpi (forboth females andmalesAst-IL6KOandfloxedmice) and46dpi (females). Numberofmicepergroup as indicated. ‹ p< 0.05 betweenAst-IL6KOandfloxedmice. (C) Representative sections from femalemice showing infiltrates (arrows) in spinal cord stainedwithCresyl violet (top) andCD3 (middle) innon-immunized(control)andimmunized(20dpi)floxedmice. Thebottompanelshows thedemyelinationof the spinal cordas revealedbyLuxolFastBlue (LFB) staining followingcolor deconvolution.All imagesatmagnification100ˆ. 3.3. ReducedGliosis andVasogenesis inSpinalCordandBrainofEAE-InducedAst-IL-6KOFemales GliosiswasevaluatedbyGFAP(astrocytes)andlectin (microglia) staining in thespinalcordof theEAE-inducedmice (Figure3A,B,D).TheareaoccupiedbyGFAPimmunostainingwasmeasured inboth thegreyandwhitematter (Figure3A).Asignificantdecrease inGFAP immunostaining in greymatterwas observed inAst-IL-6KO femalemice compared tofloxed femalemice at 46dpi, accompaniedbya less reactivemorphologyofastrocytes (Figure3D, top, left inset). Nogenotypic difference inGFAPimmunostainingwasobservedfor femalesormales ingreymatterat20dpior in thewhitematter for femalesandmalesat the timesstudied. Regardingmicrogliosis inspinal cord (Figure3D,bottom),quantificationof theoccupiedarea didnotshowsignificantdifferencesbetweengenotypes (datanotshown).Asquantificationof lectin staining isnotable toseparatemicroglia frombloodvessels, thenumberofmicrogliawascounted. Nosignificantgenotypicdifference in thenumberofmicrogliawasobserved for either femalesor 23