Bradford, Barry

Mabbott, Neil

Wijaya, Christianus

English

Edinburgh Research Explorer

     Edinburgh Research Explorer                                      Discrimination of prion strain targeting in the CNS via reactiveastrocyte heterogeneity in CD44 expressionCitation for published version:Bradford, B, Wijaya, C & Mabbott, N 2020, 'Discrimination of prion strain targeting in the CNS via reactiveastrocyte heterogeneity in CD44 expression', FENS 2020 Virtual Forum, 11/07/20 - 15/07/20.Link:Link to publication record in Edinburgh Research ExplorerDocument Version:Publisher's PDF, also known as Version of recordGeneral rightsCopyright for the publications made accessible via the Edinburgh Research Explorer is retained by the author(s)and / or other copyright owners and it is a condition of accessing these publications that users recognise andabide by the legal requirements associated with these rights.Take down policyThe University of Edinburgh has made every reasonable effort to ensure that Edinburgh Research Explorercontent complies with UK legislation. If you believe that the public display of this file breaches copyright pleasecontact openaccess@ed.ac.uk providing details, and we will remove access to the work immediately andinvestigate your claim.Download date: 23. Jul. 2021Discrimination of prion strain targeting in the CNS via reactive astrocyte heterogeneity in CD44 expressionBarry M. Bradford, Christianus A. W. Wijaya & Neil  A. MabbottThe Roslin Institute and R(D)SVS the University of Edinburgh, UK IntroductionPrion diseases result in progressive accumulation of misfolded prion protein in the central nervous system, vacuolar spongiformchange, neurodegeneration and reactive glial responses and are ultimately fatal. Many different prion agent strains exist whichdiffer in disease duration, clinical signs and targeting of neuropathology. We identified that the transmembrane glycoprotein CD44is highly upregulated in subsets of reactive astrocytes in regions of the brain specifically targeted by prions. The pattern of CD44upregulation was unique for each combination of prion agent strain and host genotype and suggest an early and critical role forastrocytes in neurodegeneration.MethodsThe distribution of CD44 was identified by immunohistochemistry in formalin fixed paraffin embedded terminal mouse brainsamples from 15 distinct prion agent strains in both Prnp-a (C57Bl6/Dk) and Prnp-b (VM/Dk) genotype mice sourced from TheRoslin Institute archive. For comparison and strain confirmation, spongiform vacuolation pattern (lesion profile), prion proteindeposition (PrPd), reactive microglial (AIF1) and astrocyte (GFAP) IHC were also performed on serial sections.A Immunohistochemical analysis using N-terminal pan CD44 antibody clone IM7 reveals minimal staining in normal uninfected mouse brain but significant upregulation of CD44 following prion infection. DAB stainingwere false coloured using a 16 colour LUT following Haematoxylin-DAB deconvolution in Image J to represent staining intensity. B False-coloured panCD44 staining from a wide array of prion strains, including naturaland experimental scrapie, Bovine spongiform encephalopathy (BSE) and Chronic wasting disease (CWD) in different Prnp genotype hosts reveals a unique pattern for every combination. C CD44 upregulation isobserved at the earliest signs of neuropathological change in the primary target area (thalamus) and progresses throughout disease. D Immunohistochemical analysis using the splice variant 6 (CD44v6) antibodyclone 9A4 confirms the astrocytic source of CD44 and upregulated expression of specific CD44 splice variants following prion infection. E Co-immunolocalisation of CD44v6, GFAP and disease-associated prionprotein reveals CD44 expressing astrocytes act as reservoirs for prion replication in the central nervous system during prion infection.Conclusions• Astrocyte heterogeneity revealed by differential expression of CD44 and CD44 splice variants during prion infection• Early and critical role for astrocyte response in neuroinflammation and neurodegenerative diseases• CD44+ astrocytes reveal targeting of prion strains allowing complete identification of both prion strain and host genotype• CD44+ astrocytes reported in Alzheimer’s, Amyotrophic lateral sclerosis and Alexander’s disease patients and is a potential biomarker for defining differential targeting (sub-type diagnosis) in human neurodegenerative diseasesResultsFront. Cell. Neurosci. doi: 10.3389/fncel.2019.00411CD44v6 PrPdGFAP MergeCD44(pan)Uninfected C57Bl/Dk ME7 Scrapie in C57Bl/Dk  22F Scrapie in C57Bl/DkME7 Scrapie in VM/DkBSE prion in C57Bl/DkCWD prion in VM/DkCD44 false colourCD44 false colour 22A Scrapie in C57Bl/Dk22A Scrapie in VM/DkCD44v622F Scrapie in C57Bl/Dk22A Scrapie in C57Bl/DkA BCD ECD4480 days post ME7 Scrapie in C57Bl/Dk  118 days post ME7 Scrapie in C57Bl/Dk  Terminal (160 days)GFAP CD44 CD44GFAP

Discrimination of prion strain targeting in the CNS via reactive astrocyte heterogeneity in CD44 expression

https://www.research.ed.ac.uk/files/205023214/2020_FENS.pdf

Discrimination of prion strain targeting in the CNS via reactive astrocyte heterogeneity in CD44 expression

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