Global gene network in STR of PQ, PY and MNPQ mouse models of PD.

<p>The genes from the most significant network of each individual pesticide mouse model were overlayed to generate global gene networks in the STR. Down-regulated genes are highlighted in green, up-regulated genes are highlighted in red. The focus molecules- G protein coupled receptors (GPCRs) members and <i>Pparδ</i> are down-regulated while <i>Atxn1</i> is up-regulated.</p

Overlapping signaling pathways between human PD transcriptome studies and signaling pathways in STR of PQ, PY and MNPQ mouse models of PD.

<p>All human IPA analysis were performed by <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036191#pone.0036191-Sutherland1" target="_blank">[18]</a>.</p><p><b>x</b> = Common Pathways.</p><p>SN: Substantia nigra, BA9: Brodman area 9, SFG: Superior frontal gyrus, L SN: Lateral substantia nigra, M SN: Medial substantia nigra, PU: Putamen, OC: Occipital cortex.</p

Common genes in the STR between PQ, PY and MNPQ mouse models of PD.

<p>A) Venn diagram of 500 up-regulated STR transcripts from PQ, PY and MNPQ models of PD. B) Venn diagram of 500 down-regulated STR transcripts from PQ, PY and MNPQ models of PD. C) Common up-regulated genes in the STR of PQ, PY and MNPQ models of PD. D) Common down-regulated genes in the STR of PQ, PY and MNPQ models of PD.</p

Venn diagram of top ten signaling pathways in PQ, PY and MNPQ models of PD.

<p>A) Signaling pathways in VMB. B) Signaling pathways in STR. C) Common overlapping pathways in VMB and STR are underlined. Considering the top ten canonical pathways there are six pathways common in VMB of PQ, PY and MNPQ mouse models. There are three pathways common in STR of PQ, PY and MNPQ mouse models. All the three common canonical pathways in STR overlap with pathways common in the VMB between the three pesticide models.</p

Validation of RNA-Seq gene expression fold changes by Taqman real time PCR.

<p>A) Gene expression fold changes in VMB. B) Gene expression fold changes in the STR.</p

Global gene network in VMB of PQ, PY and MNPQ mouse models of PD.

<p>The genes from the most significant network of each individual pesticide mouse model were overlayed to generate global gene networks in the VMB. Down-regulated genes are highlighted in green, up-regulated genes are highlighted in red. The focus molecules-G protein coupled receptors (GPCRs) members, Kelch-like ECH-associated protein 1 (<i>Keap1</i>) and peroxisome proliferator-activated receptor delta (<i>Pparδ</i>) are down-regulated in VMB.</p

Taqman assays used for quantitative RT-PCR.

<p>Taqman assays used for quantitative RT-PCR.</p

Pesticide induced loss of TH+ neurons and increased α-synuclein immunoreactivity.

<p>A) Immunostaining of SN of PQ, PY, MNPQ and wild type (WT) mice with tyrosine hydroxylase antibody. B) Percent loss of TH+ neurons and fold increase of α-synuclein immunoreactivity compared to wild type control mice.</p

Characterization of enlarged TH-positive axon terminals of <i>Atg7</i> cKO mice.

<p>(<b>A</b>) Enlarged axon terminals in the striatum of 2-month-old <i>Atg7</i> cKO mice were positive for the axon terminal proteins of midbrain DA neurons. Enlarged axon terminals (arrows in green) in <i>Atg7</i> cKO mice were stained with VMAT2 (arrows in red). Bars, 10 µm. (<b>B</b>, <b>C</b>) Enlarged axon terminals in 2-month-old <i>Atg7</i> cKO mice are Ub- and p62-negative. Enlarged axon terminals (arrows in green) in the striatum of <i>Atg7</i> cKO mice were not stained with the markers for protein inclusions such as Ub (red in ‘B’) and p62 (red in ‘C’), suggesting that they are distinct from the inclusions seen in the cell somas of <i>Atg7</i> cKO mice (<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003845#pgen-1003845-g001" target="_blank">Figure 1D</a>). Bars, 10 µm. (<b>D</b>) Ultrastructural analysis of dopaminergic axon terminals in the striatum of 3-month-old <i>Atg7</i> cWT or <i>Atg7</i> cKO mice by immunoelectron microscopy with an antibody to TH. Red circles indicate the gold particle-conjugated anti-TH antibody. Bars, 200 nm. (right) Quantification of the size distribution of axon terminals in striatal sections. Each dot (•) represents approximately 1.6% of the total axon terminal number. n = 611 terminals for cWT and 592 terminals for cKO sections.</p

Characterization of PI3K/mTOR pathways in TH-positive DA neurons of <i>Atg7</i> and/or <i>Pten</i> cKO mice.

<p>PI3K/mTOR pathway signaling, in terms of accumulation of phospho-AKT (S473), phospho-TOR (S2448), and phospho-S6 (S235/236) as indicated (in red) were unchanged in the TH-positive (green) midbrain DA neurons of <i>Atg7</i> cKO mice (C), whereas these markers were increased in TH-positive DA neurons of <i>Pten</i> cKO and <i>Atg7/Pten</i> double cKO mice (arrows in B and D). (<b>A</b>) Control cWT mice, (<b>B</b>) <i>Pten</i> cKO mice, (<b>C</b>) <i>Atg7</i> cKO mice, and (<b>D</b>) <i>Atg7/Pten</i> double cKO mice. Scale bars, 10 µm.</p