USING ZEBRAFISH AS A MODEL SYSTEM FOR DYT1 DYSTONIA

摘要: Dystonia is characterized by sustained involuntary muscle contractions producing repetitive twisting movements and abnormal postures. DYT1 dystonia, an early-onset primary caused a trinucleotide deletion in the TOR1A gene, resulting loss of single glutamic acid TorsinA protein. It unknown how this mutation causes dysfunction CNS motor circuits dystonia. The aims work were: (i) characterize zebrafish homolog human order to elucidate functions Torsins vivo; (ii) generate transgenic models dystonia suitable for mechanistic drug discovery studies. An ancestral tor1 gene found genomes several fish species was duplicated at root tetrapod lineage. In zebrafish, expressed as two isoforms with unique 5' exons. amino sequences both Torsin1 are 59% identical 78% homologous TorsinA. A novel antibody generated against Torsin1, immunoreactivity detected broadly neurons. Introduction ATP-hydrolysis abrogating mutations Walker B domain relocalization protein from endoplasmic reticulum nuclear envelope vitro, similar findings Transient knockdown expression during embryonic early larval development did not produce detectable cellular or behavioral phenotype, suggesting that essential occur later development, compensatory provided other Torsin family proteins. Co-expression dystonia-associated mutant relocalize envelope, strongly interact. view interaction, proposed dominant-negative mechanism whereby clinical disease, we stable which dystonia-related TorsinA[ΔE] neurons CNS. These animals exhibited transient, juvenile-onset hypokinetic beginning around one month lasting approximately week. Future studies using these will aim physiological molecular basis phenotype its relation