Accelerated Axonal Loss Following Acute CNS Demyelination in Mice Lacking Protein Tyrosine Phosphatase Receptor Type Z

• DOI: 10.1016/j.ajpath.2012.07.011
• Publication Date: 2012-08-31T06:27:11Z
• Authors: Jeffrey K. Huang; Carina C. Ferrari; Glaucia Monteiro ...; David Lafont; Chao Zhao; Paola Zaratin; Sandrine Pouly; Beatrice Greco; Robin J.M. Franklin
• Keywords: Remyelination

Protein tyrosine phosphatase receptor type Z (Ptprz) is widely expressed in the mammalian central nervous system and has been suggested to regulate oligodendrocyte survival differentiation. We investigated role of Ptprz remyelination after acute, toxin-induced demyelination null mice. found neither obvious impairment recruitment precursor cells, astrocytes, or reactive microglia/macrophage lesions nor a failure for cells differentiate remyelinate axons at lesions. However, we observed an unexpected increase number dystrophic by 3 days demyelination, followed prominent Wallerian degeneration 21 Ptprz-deficient Moreover, quantitative gait analysis revealed deficit locomotor behavior mutant mice, suggesting increased vulnerability axonal injury. propose that necessary maintain integrity demyelinating environment may be important target protection inflammatory diseases, such as multiple sclerosis periventricular leukomalacia. 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Gréco Zaratin like zeta modulation sensorimotor responses noxious stimuli: evidences knockout studies.Behav 2009; 201: 29-40Crossref (19) Briefly, carbonic anhydrase fibronectin III replaced LacZ reporter stop codons. WT littermates (Ptprz+/+) used controls. All surgical procedures performed under UK Home Office Project license. Focal was (Sigma-Aldrich Company Ltd, Dorset, UK) saline funiculus (Ptprz+/+). CatWalk previously.16Patrignani Lafont D.T. Muzio V. Hooft van Huijsduijnen P.F. Characterization H1 animal local systemic inflammation.J Inflamm (Lond). 2010; 16Crossref (7) The animals sacrificed 3, 5, 10, surgery analysis. Mice (n = 10 per time) perfused 4.0% paraformaldehyde. Tissues kept hours fixative, cryoprotected 30% sucrose, frozen isopentane. Sections (12 μm thick) cut cryostat −80°C until use. For immunofluorescence, sections dried washed PBS 0.3% Triton. After blocking normal donkey serum, incubated antibodies: rabbit anti-Olig2 (1:1000; Millipore, Billerica, MA) anti-NF200 Abcam, Cambridge, overnight 4°C. anti-APC/CC1 (Ab-7) (1:300; Calbiochem, San Diego, CA) anti-amyloid (APP) (clone 22C11) (1:400; Millipore) immunostaining, antigen retrieval (Dako Real Target Retrieval Solution; Dako, Glostrup, Denmark) (Vector Laboratories, Peterborough, according manufacturer instructions before antibody incubation. death analysis, TUNEL staining Situ Death Detection Kit, POD (Roche Diagnostics, Indianapolis, IN), instruction 2 washes 1× PBS, secondary antibody: Donkey (Dk) anti-mouse Cy2, Dk anti-rabbit Cy3, anti-rat Cy3 (all Jackson Immunoresearch Europe Suffolk, UK), Alexa 488 (Invitrogen, Paisley, UK). Axio Observer microscope (Carl Zeiss, Peabody, MA). Animals resin embedding group) 2.0% glutaraldehyde (pH 7.4). dissected, segments left same fixative postfixed osmium tetroxide (Oxkem Limited, Reading, embedded (TAAB Labs, Aldermaston, (1 Leica (Milton Keynes, RM2065 ultramicrotome stained 1% toluidine blue. Ultr-thin Hitachi (Maidenhead, H-600 electron microscope. Images photographed Observed Zeiss). taken 20× 40× magnification. Positive counted area. Six each group, counted. area calculated Axiovision software version 4.7.1 Results mean ± SEM. Statistical one-way variance post hoc Bonferroni's comparison test unpaired t-test. data Prism GraphPad program 5.0b (GraphPad Software Inc, CA). P < 0.05 considered statistically significant samples. To determine whether plays (OPC) differentiation, produced Ptprz−/− Ptprz+/+ (Figure 1A). Immunostaining (dpl). Nkx2.2 marker recruited OPCs, CC1 mature oligodendrocytes, Olig2 pan-oligodendrocyte marker.17Huang Jarjour A.A. Nait Oumesmar Kerninon Williams Krezel Kagechika H. 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