Journal: Neurobiology of disease

Article Title: Genetic ablation of Gpnmb does not alter synuclein-related pathology.

doi: 10.1016/j.nbd.2021.105494

Figure Lengend Snippet: Fig. 3. Gpnmb knockout does not alter recovery from lysolecithin-induced demyelination. (A) Schematic of lysolecithin demyelination/remyelination model. dpi: days post injection (B) Gpnmb IHC of corpus callosum in wildtype and knockout animals. (C–F) IHC for activated microglia (Cd68), astrocytes (Gfap), degraded myelin (dMBP) and apoptotic cells (cleaved caspase 3). Each point represents the average of up to 3 sections per animal. Integrated intensity averaged over (corpus callosum) area quantified for C-E, number of positive cells per area for F. No significant differences were observed for any of these signals when comparing between knockout and wildtype animals at any of the timepoints (t-test P > 0.05). F′, F′′, zoom in on very rare cleaved caspase 3+ signal, which was not significantly enriched in knockout relative to wildtype and in all cases far below the roughly 100 cells/mm2 previously reported using this assay in cuprizone-treated MerTK knockout mice (Shen et al., 2021). Scale bar in panel C applies as well to D and E. Scale bar in F′ applies as well to F′′.

Article Snippet: Target Figure Vendor Catalog number Link Human GPNMB 2G LS Bio LS-C340379 https://www.lsbio.com/antibodies/gpnmb-antibody-osteoactivin-antibody-clone-oti8a10-ihc-wb-western-ls-c3 40379/350990 Mouse GPNMB 2C-D, 3B, 4B R&D Systems AF2330 https://www.rndsystems.com/products/mouse-osteoactivin-gpnmb-antibody_af2330 Iba1 2G Abcam ab178846 https://www.abcam.com/iba1-antibody-epr16588-ab178846.html Mouse Cd68 2D, 3D, 4B, 4C BioRad MCA1957 https://www.bio-rad-antibodies.com/monoclonal/mouse-cd68-antibody-fa-11-mca1957.html?f=purified Mouse GFAP 2D, 3C, 4B, 4C DAKO Z0334 https://www.agilent.com/en/product/immunohistoch emistry/antibodies-controls/primary-antibodies/glial-fibrillary-acidic-protein-(concentrate)-76683 Mouse dMBP 3E Millipore AB5864 https://www.emdmillipore.com/US/en/product/Anti-Myelin-Basic-Protein-Antibody,MM_NF-AB5864 mouse cleaved caspase 3 3F Cell Signaling 9661S https://www.cellsignal.com/products/primary-antibodies/cleaved-caspase-3-asp175-antibody/9661 p129-αSyn 4B, 4C Wako 010–26,481 https://labchem-wako.fujifilm.com/us/product/detail/W01W0101-2648.html NeuN 4C Millipore MAB377B https://www.emdmillipore.com/US/en/product/Anti-NeuN-Antibody-clone-A60-biotin-conjugated,MM_N F-MAB377B Mouse Th 5C Pel Freez p40101 https://www.pelfreez-bio.com/wp-content/uploads/2014/07/74075-PDS-P40101-Tyrosine-Hydroxylase-Antibod y-Rabbit-Rev-02.pdf Human GPNMB S2A R&D Systems AF2550 https://www.rndsystems.com/products/human-osteoactivin-gpnmb-antibody_af2550 Human TH S2B Millipore AB152 https://www.emdmillipore.com/US/en/product/Anti-Tyrosine-Hydroxylase-Antibody,MM_NF-AB152 Human GPNMB RNA S2B Advanced Cell Diagnostics 413,528 https://acdbio.com/search/site/%252A413528%252A/cms/probes GPNMB knockout Gpnmbtm1.1(KOMP)Vlcg Human SNCA transgenic A53T α-synuclein transgenic line M83 Control Parkinson’s Total number of subjects 10 9 Mean Age at Death (Standard Deviation) 80.1 (12.1) 73 (6.8) Sex 4 female, 5 male, 1 unknown 7 male, 2 unknown R. Brendza et al. Neurobiology of Disease 159 (2021) 105494

Techniques: Knock-Out, Injection