Schematic illustration of one trial of the monetary incentive delay (MID) task performed by subjects in the MRI scanner.

<p>Schematic illustration of one trial of the monetary incentive delay (MID) task performed by subjects in the MRI scanner.</p

BOLD signal parameter estimates from anatomical regions during gain anticipation in different (68-bp VNTR) prodynorphin promoter polymorphism genotypes (LL-, LH-, or HH-alleles).

<p>The HH (“high level pDYN expression”) group shows significantly higher activation (see *) in the mOFC compared to the LL (“low level pDYN expression”) and LH groups.</p

Whole brain activation of all 71 participants for reward and loss anticipation conditions.

<p>(<b>a</b>) The comparison between gain and neutral anticipation (GA>NA) conditions and between loss and neutral anticipation (GA>LA) conditions showed activation in bilateral striatum, amygdala, insula, midbrain, anterior cingulate cortex and cerebellum in both incentive conditions relative to the neutral no outcome control condition. (<b>b</b>) The contrast between gain anticipation and loss anticipation (GA>LA) revealed activation in ventral striatum, ventro-medial prefrontal cortex VMPFC, thalamus and posterior cortex (PC). The threshold is p = 0.05, corrected for multiple comparisons on the voxel level (see methods).</p

The PPI analysis during gain anticipation shows higher functional coupling of the anterior mOFC seed region of the HH group with the ventral striatum (a), subgenual anterior cingulate cortex and VMPFC (b) regions compared to the LL genotype group.

<p>The threshold is p = 0.05 cluster level corrected for multiple comparisons (see methods).</p

Ten Rare, Non-synonymous Variants Shared by Individuals IV:11 and IV:18 of Family PARK_0005.

<p>The rare variants common to the two affected individuals were genotyped in 975 cases and 1014 controls. Penetrance with regard to the PD phenotype was assessed in 6 family members belonging to the same generation as the affected individuals. EA = European American.</p

Qualitative multifactorial interaction network of <i>PLXNA4</i> and genetic factors with known and hypothetical relevance to PD.

<p>Edges obtained from CIDeR are highlighted in blue, PD-specific pathways from KEGG are given in green, red edges denote annotations from OMIM and edges extracted from literature, protein-protein interaction databases or high-confidence predictions are colored black. Undirected protein-protein interactions hold circular ends, directed molecular relations are marked by arcs, whereas general regulations have arrows with no filling, activations have filled arrows and inhibitions have blunted end. Dashed lines indicate indirect effects.</p

Assessment of cell viability and subcellular protein localization in fibroblasts.

<p>(A) The presence of <i>PLXNA4</i> p.Ser657Asn do not affect cell viability as assay by live-dead staining and FACS. (B) Immunohistochemistry shows similar subcellular localization of <i>PLXNA4</i> (anti-PLXNA4, Sigma, 1∶500) in fibroblasts with and without the p.Ser657Asn amino acid substitution (scale bar = 50 µm).</p

Pedigree and Linkage Analysis.

<p>(A) Pedigree of family used for exome sequencing. Open symbols indicate unaffected family members, affected individuals are denoted by closed symbols. An arrow denotes the individuals whose exomes were sequenced. Sex was obscured and birth order was altered to protect privacy. A diagonal line indicates a deceased individual. (B) 25 genomic regions on 12 chromosomes with logarithm of the odds (LOD) score≥0.5 were identified by linkage analysis. Green boxes represent genomic regions with LOD≥0.5, yellow stars represent the location of the four candidate genes remaining after frequency assessment (<i>GOLGA4</i>-chr3, <i>PLXNA4</i>-chr7, <i>OGN</i>-chr9, <i>CPNE1</i>-chr20). <i>PLXNA4</i> on chromosome 7 represents the only of the four genes overlapping a genomic region with LOD≥0.5.</p

Metformin reverses TRAP1 mutation-associated alterations in mitochondrial function in Parkinson's disease

The mitochondrial proteins TRAP1 and HTRA2 have previously been shown to be phosphorylated in the presence of the Parkinson’s disease kinase PINK1 but the downstream signalling is unknown. HTRA2 and PINK1 loss of function causes parkinsonism in humans and animals. Here, we identified TRAP1 as an interactor of HTRA2 using an unbiased mass spectrometry approach. In our human cell models, TRAP1 overexpression is protective, rescuing HTRA2 and PINK1-associated mitochondrial dysfunction and suggesting that TRAP1 acts downstream of HTRA2 and PINK1. HTRA2 regulates TRAP1 protein levels, but TRAP1 is not a direct target of HTRA2 protease activity. Following genetic screening of Parkinson’s disease patients and healthy controls, we also report the first TRAP1 mutation leading to complete loss of functional protein in a patient with late onset Parkinson’s disease. Analysis of fibroblasts derived from the patient reveal that oxygen consumption, ATP output and reactive oxygen species are increased compared to healthy individuals. This is coupled with an increased pool of free NADH, increased mitochondrial biogenesis, triggering of the mitochondrial unfolded protein response, loss of mitochondrial membrane potential and sensitivity to mitochondrial removal and apoptosis. These data highlight the role of TRAP1 in the regulation of energy metabolism and mitochondrial quality control. Interestingly, the diabetes drug metformin reverses mutation-associated alterations on energy metabolism, mitochondrial biogenesis and restores mitochondrial membrane potential. In summary, our data show that TRAP1 acts downstream of PINK1 and HTRA2 for mitochondrial fine tuning, whereas TRAP1 loss of function leads to reduced control of energy metabolism, ultimately impacting mitochondrial membrane potential. These findings offer new insight into mitochondrial pathologies in Parkinson’s disease and provide new prospects for targeted therapies

Study Design.

<p>Study Design.</p