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Combined transcriptomics and proteomics analysis unveiled the impact of vitamin C in modulating specific mitochondrial protein abundance in the mouse liver

ABSTRACT: : In the present study, we used the Gulo-/- female and male mice model which depends entirely on ascorbate derived from the diet. We tested different concentrations of ascorbate ranging from 0 to 0.4% (w/v) ascorbate, added into drinking water, until the age of four months. Importantly, the levels of ascorbate found in the liver of Gulo-/- mice reflect the amounts of ascorbate provided in drinking water. We performed label-free Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) global quantitative proteomic profiling to identify and quantify proteins in whole liver extracts from our different ascorbate treated cohorts of Gulo-/- females and males. We compared the proteomic profiles to the transcriptomic profiles of the same treated animals. Although several proteins of the mitochondrial complex III significantly correlated with vitamin C concentrations, their corresponding transcripts did not correlate with vitamin C in both females and males. Such observations were confirmed by western blot and quantitative RT-PCR analyses, respectively. Our findings suggest that transcriptome profiling alone provides an incomplete picture of molecular changes associated with vitamin C deficiency in the liver of mice and examination of changes in protein abundances is essential to unveil variations that are not transcriptionally regulated.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Mus Musculus (ncbitaxon:10090)

SUBMITTER: Michel Lebel

PROVIDER: MSV000092040 | MassIVE | Fri May 26 08:28:00 BST 2023

SECONDARY ACCESSION(S): PXD042513

REPOSITORIES: MassIVE

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Combined transcriptomics and proteomics analysis unveiled the impact of vitamin C in modulating specific mitochondrial protein abundance in the mouse liver.

Project description:Vitamin C (ascorbate) is a crucial antioxidant and an essential cofactor of biosynthetic and regulatory enzymes. Unlike humans, mice can synthesize vitamin C thanks to the key enzyme gulonolactone oxidase (Gulo). In the present study, we used the Gulo-/- mouse model, which cannot synthesize their own ascorbate to determine the impact of this vitamin on both the transcriptomics and proteomics profiles in the liver. The study included Gulo-/- mouse groups treated with either sub-optimal or optimal vitamin C concentrations in drinking water. We found a distinctive difference in the mRNA and protein profiles as a function of sex between all the mouse groups. Despite this sexual dimorphism, Spearman correlation analyses were conducted to divulge which transcripts and proteins correlated with hepatic vitamin C concentrations in both females and males. Such analysis on transcriptomics data from females and males revealed changes in a wide array of biological processes involved in glucose, lipid, steroid, and glutathione metabolisms as well as in acute-phase response. Furthermore, although several proteins of the mitochondrial complex III significantly correlated with vitamin C concentrations, their corresponding transcripts did not correlate with vitamin C in both females and males. Thus, this side-by-side comparison between transcriptome and proteome supported the view that post-transcriptional processes play a major role in the regulation of cellular respiration in the liver upon vitamin C deficiency. Our findings suggest that transcriptome profiling alone provides an incomplete picture of molecular changes associated with vitamin C deficiency in the liver of mice and examination of changes in protein abundances is essential to unveil variations that are not transcriptionally regulated.

2023-06-01 | GSE233598 | GEO

Vitamin C modulates the levels of several proteins of the mitochondrial complex III and its activity in the mouse liver

Project description:In the present study, we used the Gulo-/- female and male mice model which depends entirely on ascorbate derived from the diet. Importantly, the levels of ascorbate found in the serum of Gulo-/- mice reflect the amounts of ascorbate provided in drinking water. We tested different concentrations of ascorbate ranging from 0 to 0.4% (w/v) ascorbate, added into drinking water, until the age of four months. We performed label-free Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) global quantitative proteomic profiling to identify and quantify proteins in microsomal enriched liver extracts (MEE) from our different ascorbate treated cohorts of Gulo-/- females and males. We found that the MS intensities of several proteins in the mitochondrial complex III of the electron transport chain correlated positively with liver ascorbate concentrations in both Gulo-/- females and males. Consequently, the mitochondrial complex III activity in Gulo-/- female and male mice treated with suboptimal hepatic concentrations of ascorbate was significantly lower than Gulo-/- mice treated with optimal ascorbate concentration. Finally, proteins involved in complement activation correlated negatively with liver ascorbate concentrations in both Gulo-/- males and females.

2022-07-13 | MSV000089881 | MassIVE

Vitamin C differentially impacts the serum proteome profile in female and male mice

Project description:Suboptimal blood vitamin C (ascorbate) level increases the risk of several chronic diseases. However, the detection of hypovitaminosis C is not a simple task as ascorbate is unstable in blood samples. In this study, we examined the serum proteome of mice lacking the enzyme gulonolactone oxidase (Gulo) required for the biosynthesis of ascorbate. Gulo-/- mice were supplemented with different concentrations of ascorbate in drinking water and serum was collected to identify proteins correlating with serum ascorbate levels using an LC-MS/MS quantitative proteomic approach. Parallel reaction monitoring was performed to validate the correlations. We uncovered that the serum proteome profiles differ significantly between male and female mice. Also, unlike Gulo-/- males, a four-week treatment of ascorbate did not entirely re-establish the serum proteome profile of ascorbate-deficient Gulo-/- females to the optimal profile exhibited by Gulo-/- females that never experienced an ascorbate deficiency. Finally, serum proteins involved in retinoid metabolism, cholesterol, and lipid transport were similarly affected by ascorbate levels in males and females. In contrast, proteins regulating serum peptidases and protein of the acute phase response were different between males and females. These proteins are potential biomarkers correlating with blood ascorbate levels and require further study in standard clinical settings.

2021-06-28 | MSV000087709 | MassIVE

Vitamin C (ascorbate) modulates the levels of several proteins in the brain of mice

2023-06-07 | MSV000092125 | MassIVE

Vitamin C (ascorbate) modulates the levels of several proteins in the heart of mice

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Vitamin C (ascorbate) modulates the levels of several proteins in the spleen of mice

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Brain-synthetized IGF-1 sexual dimorphism and proteostasis

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2022-09-14 | PXD028458 | JPOST Repository

Serum proteomics reveals a sexually dimorphic effect of vitamin D status correction on proteins of the blood coagulation pathway among overweight adults

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Vitamin C alters the amount of specific endoplasmic reticulum associated proteins involved in lipid metabolism in the liver of mice synthesizing a nonfunctional WRN mutant protein

Project description:Werner syndrome (WS) is a premature aging disorder caused by mutations in a DNA helicase/exonuclease. Mice lacking the helicase domain of the Wrn protein orthologue exhibit transcriptomic and metabolic alterations, some of which are reversed by vitamin C. Recent studies on the liver of these animals indicated that the Wrn mutant protein is associated with the endoplasmic reticulum (ER) resulting in an ER stress response. In this study, we identified liver proteins that exhibit actual level differences in the ER fraction between wild type and Wrn mutant mice using quantitative proteomic profiling with label-free Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS). Multiple Reaction Monitoring (MRM) and immunoblotting were performed to validate findings in a secondary independent cohort of wild type and Wrn mutant mice in the presence or absence of vitamin C in drinking water. The list of identified proteins showing significant altered expression levels was compared to transcriptomic results that were obtained in previous studies to assess the extent of correlation between the mRNA levels and their corresponding encoded protein levels in the liver of our different mouse cohorts.

2018-03-02 | PXD007758 | Pride

Nutritional effects by beta-carotene in liver in males and females of control wildtype mice versus BCMO knockout mice

Project description:Molecular mechanisms triggered by high dietary beta-carotene (BC) intake in liver are largely unknown. We performed microarray gene expression analysis on liver tissue of BC supplemented beta-carotene 15,150-monooxygenase 1 knockout (Bcmo1-/-) mice, which are—like humans—able to accumulate BC. This was compared with litter mates being wild-type (Bcmo1+/+) mice, and we analysed both males and females, as we previously showed that in lung tissue we observed opposite gene regulation between males and females (Van Helden et al., CMLS 2011).

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