Bright-field immunohistochemical stainings were performed in age-matched WT-F1-hybrids, PrPmtA and DM mouse (for Pink1 2 / 2, see Supplementary Material, Fig.

Immunohistochemistry in midbrain at ages 15вЂ“17 months. In high-magnification panels (panel row below), the granular and thread-like composition of the signals and the corkscrew-appearance of some immunopositive neurites become apparent (3J, black arrows), as well as the higher sensitivity of the pSer129-SNCA antibody for this pattern. In DM, strong immunoreactivity throughout the gray substance in neuronal cytoplasm with extension along neurites can be observed. In PrPmtA, the pSer129-SNCA antibody detected few neuronal nuclei owing to the overexpression of SNCA (3D, open arrow). In WT spinal cord at old age, no protein aggregates were detected. Protein aggregation was detected by antibodies against phospho-Serine129-alpha-synuclein (pSer129-SNCA), p62 or ubiquitin. S3 upper panels) spinal cord (focused on one anterior horn with ventral surface below and midline central canal to the right) in comparison. Immunohistochemistry in spinal cord at ages 15вЂ“17 months. In comparison, DM animals ( n 1вЃ„4 51) showed some mortality early on, which increased in a steeply progressive manner after age 450 days (significant P -value, 0.0001, median survival reduced from 723 to 546 days). ( C ) WT-F1-hybrids with a 50 : 50 ratio of FVB/N and 129/SvEv background ( n 1вЃ„4 34) showed few deaths during young age owing to aggressive behavior, and a much increased lifespan as often seen after outbreeding.

The SM animals with Pink1 2 / 2 ( n 1вЃ„4 104) exhibited an extended lifespan (significant P -value 1вЃ„4 0.0159, median survival lengthened from 545 to 592 days). ( B ) WT 129/SvEv mice ( n 1вЃ„4 75) showed a moderate number of animal deaths at young age and slowly progressive mortality, as expected from an inbred line.

( A ) While male FVB/N mice (starting n 1вЃ„4 61) showed aggressive behavior at young age and inbreeding multimorbidity at advanced age resulting in constant high mortality across all ages independent from WT or mutant genotype, the homozygous PrPmtA line with overexpression of human A53T-SNCA ( n 1вЃ„4 77) resulted in some early mortality (significant P -value 1вЃ„4 0.0002 in MantelвЂ“ Cox test, median survival reduced from 372 to 226 days). Thus, our study confirmed that SNCA-triggered neurotoxicity is exacerbated by the absence of PINK1, and identified a novel molecular signature that is detectable early in the course of this double pathology. The observed downregulations for Dapk1, Dcaf17, Rab42, and the novel SNCA-marker Lect1 as well as the upregulations for Dctn5, Mrpl9, Tmem181a, Xaf1, H2afx reflect changes in ubiquitination, mitochondrial / synaptic / microtubular / cell adhesion dynamics, and DNA damage. Global brain transcriptome profiles and qPCR validation experiments identified many consistent transcriptional dysregulations already at age 6 weeks, which were absent from SM. Brain proteome quantifications of ubiquitination sites documented altered degradation of SNCA and the DNA-damage marker H2AX at age 18 months. In contrast to SM, a quarter of DM animals manifested progressive paralysis at ages >1 year and exhibited protein aggregates immunopositive for pSer129-SNCA, p62, and ubiquitin in spinal cord and basal brain. DM animals showed potentiated phenotypes in comparison to both single mutants (SM), with reduced survival and strongly reduced spontaneous movements from age 3 months onwards. We studied their lifespan and behavior, histological and molecular anomalies at late and early ages. Here, we crossed previously characterized mice with A53T-SNCA overexpression and with Pink1 deletion to generate double mutants (DM). Interactions between these triggers and their convergence onto shared pathways are crucial, but currently conflicting evidence exists. The common age-related neurodegeneration of Parkinson's disease (PD) can result from dominant causes like increased dosage of vesicle-associated alpha-synuclein (SNCA), or recessive causes like deficiency of mitophagy factor PINK1.

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