Western blot (WB) analysis of a-Synuclein protein levels.

<p>mThy1 and PDGF transgenic a-Synuclein mice and non transgenic littermates (n tg) are compared: (A, C) WB and (B, C) densitometric analysis (normalized against GAPDH) shows a significant increase in a-Synuclein expression in mThy1 mice both in the cortex (A, B, *<0.05, n = 3 replicates, data shown as means ± SEM) and in the olfactory bulb (C, D, *<0.005).</p

Effect of a-Synuclein expression level and temporal expression pattern on olfactory bulb neurogenesis.

<p>The early expression of relatively low levels of transgene (tg, purple bar) in PDGF animals leads to a significant reduction of newborn neurons by affecting cell survival but not cell differentiation. In contrast, a stronger expression of a-Synuclein at later stages of neuronal maturation in mThy1 animals causes lesser changes in adult olfactory neurogenesis mainly affecting dopaminergic differentiation in the GLOM.</p

Experimental design.

<p>Control and mThy1 a-Synuclein transgenic animals were subjected to a paradigm with 5 consecutive days of i.p. BrdU injections at the age of 4 months and histological analysis at the age of 5 months (panel A). In the SVZ no difference in the number of neuronal progenitor (PCNA-positive) cells was detectable. In the granular cell layer of the olfactory bulb the numbers of surviving BrdU-positive cells, the percentage of BrdU/NeuN double-positive cells as well as the numbers of newly generated neurons were not significantly different in both groups, indicating that mThy1 human wild-type a-Synuclein had no effect on adult granular cell layer neurogenesis (panel B). Numbers are given as means ± standard error means.</p

Comparison of olfactory bulb neurogenesis in mThy1 & PDGF transgenic mice.

<p>Stereologically quantified numbers of cell survival (BrdU-positive cells: panels A, D), percentage of neuronal / dopaminergic differentiation (BrdU/NeuN+ in panel B, respectively BrdU/TH+ cells in panel E) and new neurons (panels C, F) both in the granular cell layer (panels A, B, C) and the glomerular cell layer (D, E, F) of the olfactory bulb. Results were normalized to the numbers of corresponding control animals of each transgenic animal strain. In the granular cell layer, the number of new neurons was decreased in PDGF animals, while mThy1 animals display an unaltered neurogenesis. In the glomerular cell layer however, there was a reduced cell survival in PDGF, respectively a drop in dopaminergic differentiation in mThy1 animals causing a significant reduction in the number of new dopaminergic cells in comparison to respective control animals, which was not significantly different between both transgenic animal strains.</p

Temporal expression analysis in mThy1 transgenic animals.

<p>Coexpression analysis of human-wildtype a-Synuclein with markers of cell maturation from stem cell stage (SOX2), maturing neuroblast (DCX) and mature (Calretinin, NeuN) and dopaminergic neurons (TH). Detailed immunofluorescent analysis in the subventricular zone (Sox2, DCX), respectively olfactory bulb Granule cell layer (NeuN) and Glomerular cell layer (Calretinin, TH) reveals an expression at the mature neuron stage (Calretinin/NeuN-positive). Arrows indicate positive coexpression of human-wildtype a-Synuclein with NeuN, Calretinin and TH. Scale bar 20μm.</p

Stereological quantification in the olfactory bulb glomerular cell layer.

<p>the number of BrdU-positive cells did not differ between transgenic a-Synuclein mThy1 and control animals (panel A). The analysis of the percentage of BrdU/TH-double-positive cells revealed a non-significant (p = 0.07) trend (panel B). The numbers of newly generated dopaminergic neurons are significantly reduced in the a-Synuclein transgenic animals (p = 0.04, panel C). Immunofluorescent confocal images of the glomerular cell layer show colocalization of markers of neuronal (NeuN, panel D) and dopaminergic differentiation (TH, panel E) with BrdU, a marker of cell survival (panel F). Scale bar 20μm.</p

Additional file 13: of Sporadic Parkinson’s disease derived neuronal cells show disease-specific mRNA and small RNA signatures with abundant deregulation of piRNAs

Figure S6. Global statistics on RRBS and analysis of differential methylation. (TIF 351 kb

Additional file 7: of Sporadic Parkinson’s disease derived neuronal cells show disease-specific mRNA and small RNA signatures with abundant deregulation of piRNAs

Table S4. Differential expression analysis for mature miRNAs in fibroblasts, iPSCs/ESCs and neurons for the comparison PD vs. CTRL. (XLSX 718 kb

Additional file 9: of Sporadic Parkinson’s disease derived neuronal cells show disease-specific mRNA and small RNA signatures with abundant deregulation of piRNAs

Figure S4. Small RNA content analysis and library size distribution. (TIF 491 kb

Additional file 10: of Sporadic Parkinson’s disease derived neuronal cells show disease-specific mRNA and small RNA signatures with abundant deregulation of piRNAs

Table S6. Differential expression analysis for piRNAs/piRNA-like molecues and mature miRNAs for the comparison control fibroblasts vs. control iPSCs/ESCs and control iPSCs/ESCs vs. control neurons. (XLSX 7706 kb