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a Diagram showing that thymidine <t>phosphorylase</t> catalyzes the conversion of thymidine and deoxyuridine into their respective bases (thymine and uracil) and 2-deoxyribose 1-phosphate. b Western blot analysis of thymidine phosphorylase (TP) expression levels in brain and liver in 5 month-old G2019S LRRK2 knockin mice and controls ( n = 2 per group). c qRT-PCR analysis of TYMP levels in ventral midbrain of 5 month-old G2019S LRRK2 knockin mice and controls ( n = 4 per group). Data are presented as mean ± SEM. Unpaired two-tailed Student’s t -test was performed to evaluate the effects of G2019S LRRK2 mutation, no significant effect was found. d Volcano plot showing differential gene expression in the liver of 5 month-old G2019S LRRK2 knockin mice compared to wild-type controls. Each point represents one gene. X-axis shows log 2 (fold change) in gene expression in G2019S LRRK2 knockin mice compared to wild-type controls. Y-axis shows -log 10 (adjusted p -value) from limma’s empirical Bayes moderated analysis (eBayes) combined with Benjamini-Hochberg (BH) for multiple testing correction ( n = 4 per group). Gene expression with adjusted P <0.05 are considered significantly altered. Significantly upregulated genes are highlighted in red and significantly downregulated genes are highlighted in blue. e Volcano plot illustrating enriched biological processes in G2019S LRRK2 knockin mice compared to wild-type controls as identified by gene set enrichment analysis (GSEA). Each point represents one process. X-axis shows normalized enrichment score, Y-axis shows -log 10 (adjusted p -value), significance was defined as a Benjamini–Hochberg adjusted P <0.05 ( n = 4 per group). Significantly upregulated pathways are highlighted in red and significantly downregulated pathways are highlighted in blue. f Western blot analysis and quantification ( g-m ) of TP, total and phosphorylated LRRK2, Rab10, and Rab12 expression levels in the liver of G2019S LRRK2 knockin and wild-type mice under Con.diet and HFD conditions ( n = 3 per group). WT/WT, wild-type mice; KI/KI, homozygous G2019S LRRK2 knockin mice. Data are presented as mean ± SEM. A two-way ANOVA was performed to evaluate the effects of high-fat diet and G2019S LRRK2 mutant and their interaction followed by Fisher’s LSD comparisons. Significant main effects were found (* P <0.05, ** P <0.01).
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a Diagram showing that thymidine <t>phosphorylase</t> catalyzes the conversion of thymidine and deoxyuridine into their respective bases (thymine and uracil) and 2-deoxyribose 1-phosphate. b Western blot analysis of thymidine phosphorylase (TP) expression levels in brain and liver in 5 month-old G2019S LRRK2 knockin mice and controls ( n = 2 per group). c qRT-PCR analysis of TYMP levels in ventral midbrain of 5 month-old G2019S LRRK2 knockin mice and controls ( n = 4 per group). Data are presented as mean ± SEM. Unpaired two-tailed Student’s t -test was performed to evaluate the effects of G2019S LRRK2 mutation, no significant effect was found. d Volcano plot showing differential gene expression in the liver of 5 month-old G2019S LRRK2 knockin mice compared to wild-type controls. Each point represents one gene. X-axis shows log 2 (fold change) in gene expression in G2019S LRRK2 knockin mice compared to wild-type controls. Y-axis shows -log 10 (adjusted p -value) from limma’s empirical Bayes moderated analysis (eBayes) combined with Benjamini-Hochberg (BH) for multiple testing correction ( n = 4 per group). Gene expression with adjusted P <0.05 are considered significantly altered. Significantly upregulated genes are highlighted in red and significantly downregulated genes are highlighted in blue. e Volcano plot illustrating enriched biological processes in G2019S LRRK2 knockin mice compared to wild-type controls as identified by gene set enrichment analysis (GSEA). Each point represents one process. X-axis shows normalized enrichment score, Y-axis shows -log 10 (adjusted p -value), significance was defined as a Benjamini–Hochberg adjusted P <0.05 ( n = 4 per group). Significantly upregulated pathways are highlighted in red and significantly downregulated pathways are highlighted in blue. f Western blot analysis and quantification ( g-m ) of TP, total and phosphorylated LRRK2, Rab10, and Rab12 expression levels in the liver of G2019S LRRK2 knockin and wild-type mice under Con.diet and HFD conditions ( n = 3 per group). WT/WT, wild-type mice; KI/KI, homozygous G2019S LRRK2 knockin mice. Data are presented as mean ± SEM. A two-way ANOVA was performed to evaluate the effects of high-fat diet and G2019S LRRK2 mutant and their interaction followed by Fisher’s LSD comparisons. Significant main effects were found (* P <0.05, ** P <0.01).
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a Diagram showing that thymidine <t>phosphorylase</t> catalyzes the conversion of thymidine and deoxyuridine into their respective bases (thymine and uracil) and 2-deoxyribose 1-phosphate. b Western blot analysis of thymidine phosphorylase (TP) expression levels in brain and liver in 5 month-old G2019S LRRK2 knockin mice and controls ( n = 2 per group). c qRT-PCR analysis of TYMP levels in ventral midbrain of 5 month-old G2019S LRRK2 knockin mice and controls ( n = 4 per group). Data are presented as mean ± SEM. Unpaired two-tailed Student’s t -test was performed to evaluate the effects of G2019S LRRK2 mutation, no significant effect was found. d Volcano plot showing differential gene expression in the liver of 5 month-old G2019S LRRK2 knockin mice compared to wild-type controls. Each point represents one gene. X-axis shows log 2 (fold change) in gene expression in G2019S LRRK2 knockin mice compared to wild-type controls. Y-axis shows -log 10 (adjusted p -value) from limma’s empirical Bayes moderated analysis (eBayes) combined with Benjamini-Hochberg (BH) for multiple testing correction ( n = 4 per group). Gene expression with adjusted P <0.05 are considered significantly altered. Significantly upregulated genes are highlighted in red and significantly downregulated genes are highlighted in blue. e Volcano plot illustrating enriched biological processes in G2019S LRRK2 knockin mice compared to wild-type controls as identified by gene set enrichment analysis (GSEA). Each point represents one process. X-axis shows normalized enrichment score, Y-axis shows -log 10 (adjusted p -value), significance was defined as a Benjamini–Hochberg adjusted P <0.05 ( n = 4 per group). Significantly upregulated pathways are highlighted in red and significantly downregulated pathways are highlighted in blue. f Western blot analysis and quantification ( g-m ) of TP, total and phosphorylated LRRK2, Rab10, and Rab12 expression levels in the liver of G2019S LRRK2 knockin and wild-type mice under Con.diet and HFD conditions ( n = 3 per group). WT/WT, wild-type mice; KI/KI, homozygous G2019S LRRK2 knockin mice. Data are presented as mean ± SEM. A two-way ANOVA was performed to evaluate the effects of high-fat diet and G2019S LRRK2 mutant and their interaction followed by Fisher’s LSD comparisons. Significant main effects were found (* P <0.05, ** P <0.01).
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a Diagram showing that thymidine phosphorylase catalyzes the conversion of thymidine and deoxyuridine into their respective bases (thymine and uracil) and 2-deoxyribose 1-phosphate. b Western blot analysis of thymidine phosphorylase (TP) expression levels in brain and liver in 5 month-old G2019S LRRK2 knockin mice and controls ( n = 2 per group). c qRT-PCR analysis of TYMP levels in ventral midbrain of 5 month-old G2019S LRRK2 knockin mice and controls ( n = 4 per group). Data are presented as mean ± SEM. Unpaired two-tailed Student’s t -test was performed to evaluate the effects of G2019S LRRK2 mutation, no significant effect was found. d Volcano plot showing differential gene expression in the liver of 5 month-old G2019S LRRK2 knockin mice compared to wild-type controls. Each point represents one gene. X-axis shows log 2 (fold change) in gene expression in G2019S LRRK2 knockin mice compared to wild-type controls. Y-axis shows -log 10 (adjusted p -value) from limma’s empirical Bayes moderated analysis (eBayes) combined with Benjamini-Hochberg (BH) for multiple testing correction ( n = 4 per group). Gene expression with adjusted P <0.05 are considered significantly altered. Significantly upregulated genes are highlighted in red and significantly downregulated genes are highlighted in blue. e Volcano plot illustrating enriched biological processes in G2019S LRRK2 knockin mice compared to wild-type controls as identified by gene set enrichment analysis (GSEA). Each point represents one process. X-axis shows normalized enrichment score, Y-axis shows -log 10 (adjusted p -value), significance was defined as a Benjamini–Hochberg adjusted P <0.05 ( n = 4 per group). Significantly upregulated pathways are highlighted in red and significantly downregulated pathways are highlighted in blue. f Western blot analysis and quantification ( g-m ) of TP, total and phosphorylated LRRK2, Rab10, and Rab12 expression levels in the liver of G2019S LRRK2 knockin and wild-type mice under Con.diet and HFD conditions ( n = 3 per group). WT/WT, wild-type mice; KI/KI, homozygous G2019S LRRK2 knockin mice. Data are presented as mean ± SEM. A two-way ANOVA was performed to evaluate the effects of high-fat diet and G2019S LRRK2 mutant and their interaction followed by Fisher’s LSD comparisons. Significant main effects were found (* P <0.05, ** P <0.01).

Journal: bioRxiv

Article Title: System-wide profiling reveals metabolic alterations in a G2019S LRRK2 knockin mouse model of Parkinson’s disease: systemic depletion of pyrimidine nucleosides

doi: 10.64898/2025.12.31.697225

Figure Lengend Snippet: a Diagram showing that thymidine phosphorylase catalyzes the conversion of thymidine and deoxyuridine into their respective bases (thymine and uracil) and 2-deoxyribose 1-phosphate. b Western blot analysis of thymidine phosphorylase (TP) expression levels in brain and liver in 5 month-old G2019S LRRK2 knockin mice and controls ( n = 2 per group). c qRT-PCR analysis of TYMP levels in ventral midbrain of 5 month-old G2019S LRRK2 knockin mice and controls ( n = 4 per group). Data are presented as mean ± SEM. Unpaired two-tailed Student’s t -test was performed to evaluate the effects of G2019S LRRK2 mutation, no significant effect was found. d Volcano plot showing differential gene expression in the liver of 5 month-old G2019S LRRK2 knockin mice compared to wild-type controls. Each point represents one gene. X-axis shows log 2 (fold change) in gene expression in G2019S LRRK2 knockin mice compared to wild-type controls. Y-axis shows -log 10 (adjusted p -value) from limma’s empirical Bayes moderated analysis (eBayes) combined with Benjamini-Hochberg (BH) for multiple testing correction ( n = 4 per group). Gene expression with adjusted P <0.05 are considered significantly altered. Significantly upregulated genes are highlighted in red and significantly downregulated genes are highlighted in blue. e Volcano plot illustrating enriched biological processes in G2019S LRRK2 knockin mice compared to wild-type controls as identified by gene set enrichment analysis (GSEA). Each point represents one process. X-axis shows normalized enrichment score, Y-axis shows -log 10 (adjusted p -value), significance was defined as a Benjamini–Hochberg adjusted P <0.05 ( n = 4 per group). Significantly upregulated pathways are highlighted in red and significantly downregulated pathways are highlighted in blue. f Western blot analysis and quantification ( g-m ) of TP, total and phosphorylated LRRK2, Rab10, and Rab12 expression levels in the liver of G2019S LRRK2 knockin and wild-type mice under Con.diet and HFD conditions ( n = 3 per group). WT/WT, wild-type mice; KI/KI, homozygous G2019S LRRK2 knockin mice. Data are presented as mean ± SEM. A two-way ANOVA was performed to evaluate the effects of high-fat diet and G2019S LRRK2 mutant and their interaction followed by Fisher’s LSD comparisons. Significant main effects were found (* P <0.05, ** P <0.01).

Article Snippet: Primary antibodies used for Western blotting include: LRRK2 (75-253, NeuroMab), pS935-LRRK2 (ab133450, Abcam), pS1292-LRRK2 (ab203181, Abcam), Rab8 (D22D8, Cell Signaling Technology), pT72-Rab8a (ab230260, Abcam), Rab10 (8127S; Cell Signaling Technology), pThr73-Rab10 (ab230261, Abcam), Rab12 (18843-1-AP, Protein Tech), pSer106-Rab12 (ab256487, Abcam), Actin (MAB1501, Millipore), Thymidine Phosphorylase (AF7568, Novus Biologicals).

Techniques: Western Blot, Expressing, Knock-In, Quantitative RT-PCR, Two Tailed Test, Mutagenesis, Gene Expression