The areas in the grey boxes are shown below in higher magnification. pixel section of larval ventral nerve cord as depicted in 5A. (C) Triton x-100 fractionation of samples from control and Vps13 mutant travel heads analyzed for the levels of Triton x-100 insoluble ubiquitylated proteins. The quantification shows the mean and SEM of at least five larval ventral nerve cord stainings per condition. For statistical analysis a two-tailed students T-test was used in combination with a Welchs correction if necessary. P 0.01 is **.(TIF) pone.0170106.s003.tif (1.7M) GUID:?1345315D-1CF5-48D1-9B1F-824E3683C9FC S4 Fig: Accumulation of ubiquitylated protein puncta in the adult eye lobe. Stainings of the eye lobes of 1 1 day aged adult control, mutant and excision line 3 stained for ubiquitylated proteins, Ref(2)p and DAPI. The higher magnification pictures are of the areas in the grey boxes. Arrows indicate colocalization of Ref(2)P and Ubiquitin positive foci. The scale bar in the overview picture indicates 50 m and the scale bar in the zoom in indicates 20 m.(TIF) pone.0170106.s004.tif (5.2M) GUID:?962626A5-3721-4E4C-BEA1-5305479A7D1A S1 Table: Details of the life span experiments presented. Depicted per physique are the travel lines used, the number of flies used for the experiment, the median life span, the Mantel-Cox test (M-C test) and Gehan-Breslow-Wilcoxon test (G-B-W test).(TIF) pone.0170106.s005.tif (243K) GUID:?90EF116D-394D-45DD-A0EF-CC7B277E3E8D S1 Movie: Movie of a climbing assay conducted with the BMS-777607 control and mutant flies. (MP4) Rabbit Polyclonal to FZD6 pone.0170106.s006.mp4 (11M) GUID:?80D1311B-68AF-4665-A7AB-744D2006984F Data Availability StatementAll relevant data are within the paper and its Supporting Information files. Abstract Chorea-Acanthocytosis is BMS-777607 usually a rare, neurodegenerative disorder characterized by progressive loss of locomotor and cognitive function. It is caused by loss of function mutations in the (mutant line. The gene encoded a protein of comparable size as human VPS13A. Our data suggest that Vps13 is usually a peripheral membrane protein located to endosomal membranes and enriched in the travel head. mutant flies showed a shortened life span and age associated neurodegeneration. mutant flies were sensitive to proteotoxic stress and accumulated ubiquitylated proteins. Levels of Ref(2)P, the orthologue of p62, were increased and protein aggregates accumulated in the central nervous system. Overexpression of the human Vps13A protein in the mutant flies partly rescued apparent phenotypes. This suggests a functional conservation of human VPS13A and Vps13. Our results demonstrate that Vps13 is essential to maintain protein homeostasis in the larval and adult brain. mutants are suitable to investigate the function of Vps13 in the brain, to identify genetic enhancers and suppressors and to screen for potential therapeutic targets for Chorea-Acanthocytosis. Introduction Chorea-Acanthocytosis (ChAc, MIM 200150) is usually a BMS-777607 rare neurodegenerative disorder characterized by chorea, orofacial dyskinesia and psychiatric symptoms including tics (reviewed in [1,2]). In addition to the neurological symptoms, spiky red blood cells (acanthocytes) are often observed. ChAc is usually a recessively inherited disease caused by mutations in the gene, hereafter called [3,4]. These mutations mostly lead to absence or reduced levels of the HsVPS13A (or also called chorein) protein [5]. Symptoms manifest on average at the age of 32 [1]. The pathophysiology of ChAc is largely unknown and it is not clear why HsVPS13A loss of function leads to the symptoms presenting in ChAc patients. is usually evolutionarily conserved and orthologues are present in various organisms such as and [6C8]. HsVPS13A belongs to the VPS13 family of proteins, which in humans consists of four members, VPS13A to D. All members have an N-terminal chorein domain name of unknown function. Besides HsVPS13A other members of this family are also associated with medical conditions. mutations cause Cohen syndrome, a developmental disorder characterized by mental retardation, microcephaly and facial dysmorphisms [9]. VPS13B has been reported to be a Rab6 effector that controls Golgi integrity [10,11]. mutations have recently been described to cause autosomal-recessive early-onset Parkinsons disease, probably by alteration of mitochondrial morphology and function [12]. The VPS13C protein has also been suggested to play a role in adipogenesis [13]. Additionally, a number of genetic studies have found an association of with glucose and insulin metabolism [14,15], and of.