Pedigree ALS85

1. RNA binding proteins and neurodegeneration: TDP 43 Familial ALS M377V Chick embryo Developmental delay W W W M W M Pedigree ALS85 Transverse Section IHC Transgene expression W W W W W M M M M Apoptotic Cells Tunel Staining Wild type M337V FTD ALS/MND Sporadic ALS G294A Sporadic ALS Q331K Sreedharan et al. Science 2008

2. RNA binding proteins and neurodegeneration: FUS Anti-FUS GFP-FUS DAPI Merge Wildtype FUS FUS R514G Average age at onset 35 years Average survival 12 months Linkage to Chromosome 16 LOD score = 4 Ruddy et al. AJHG 2003 3 different missense mutations 8 unrelated familial ALS kindreds Arg 521 Cys Arg 515 Gly Arg 521 His Mutant FUS forms cytoplasmic inclusions In ALS patients Mutations cause FUS to relocate to the cytoplasm Because they disrupt their nuclear localising signal Vance et al . Science 2009

3. Toxic RNA expansion and neurodegeneration: ORF72 Le Ber et.al 2009 Gijselinck 2010 Momeni et.al 2006 Morita et.al 2006 Vance et.al 2006 Valdmanis et.al 2007 Pearson 2011 Boxer et.al 2010 ~ 4MB Overlap = ALS-FTD = ALS = FTD Multigenerational Dutch kindred ALS and FTD variable phenotype Linkage to Chr 9p LOD score = 3 Vance et al. Brain 2006 10 kindreds linked Region narrowed to 4MB BAC haploid clones DNA capture Brain RNA sequenced Whole genome Next generation seq. 97 SNP common haplotype C9ORF72 intronic hexanucleotide (GGGGCC)n expansion mutation 700 to 1,000 repeats De Jesus-Hernandez Neuron 2011 Renton et al Neuron 2011 Expanded Intronic repeat detected by PCR and Southern Blot