Untargeted Metabolomics Reveals the Function of GPRC6A in Amino Acid and Lipid Metabolism in Mice
Abstract
:1. Introduction
2. Materials and Methods
2.1. Animals and Diets
2.2. Hematoxylin–Eosin (HE) Staining
2.3. Blood Collection and Serum Separation
2.4. Biochemical Parameter Determination
2.5. Metabolite Extraction
2.6. LC-MS/MS
2.7. Data Preprocessing and Annotation
2.8. KEGG Pathway Analysis
3. Results
3.1. Effect of Dietary Protein on GPRC6A+/+ and GPRC6A−/− Mice
3.2. Loss of GPRC6A Altered Serum Biochemical Parameters
3.3. Loss of GPRC6A Altered Free Amino Acid Levels in Mice
3.4. Metabolomics Screening for Differentially Expressed Metabolites between GPRC6A +/+ and GPRC6A−/− Mice
3.5. Pathway Enrichment Analysis Based on the KEGG (Kyoto Encyclopedia of Genes and Genomes) Library
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Ingredient (% by Weight) | 10% Protein (LP) | 20% Protein (NP) |
---|---|---|
Corn Sugar | 46.06 | 39.75 |
Casein Lactic | 10.00 | 20.00 |
Granular Sugar | 11.74 | 10.00 |
Dextrin | 15.30 | 13.20 |
Solka Floc-40 | 5.00 | 5.00 |
AIN-93 Mineral Mix | 3.50 | 3.50 |
AIN-93 Vitamin Mix | 1.00 | 1.00 |
l-Cystine | 0.15 | 0.30 |
Choline Bitartrate | 0.25 | 0.25 |
Soy Oil | 7.00 | 7.00 |
Sum | 100 | 100 |
AAs (μM) | WT–NP | KO–NP | WT–LP | KO–LP | p-Value | |
---|---|---|---|---|---|---|
Genotype | Protein | |||||
His | 60.5 ± 1.8 | 59.3 ± 3 | 65 ± 2.6 | 62.2 ± 2.7 | 0.432 | 0.152 |
Arg | 114.6 ± 2.6 | 140.2 ± 3.9 ** | 146.9 ± 7.8 | 145.9 ± 7.5 | 0.059 | 0.005 |
Asn | 51.1 ± 2 | 62 ± 5.2 | 55.1 ± 2.6 | 60.5 ± 1.4 | 0.013 | 0.693 |
Gln | 480.8 ± 10.9 | 447.3 ± 19 | 556.8 ± 7.8 | 463 ± 8.7 *** | <0.001 | 0.001 |
Ser | 102.3 ± 4.4 | 129.3 ± 5.1 | 136.9 ± 4.6 | 129.7 ± 3.1 | 0.166 | 0.019 |
Gly | 249.2 ± 11.1 | 267.3 ± 8.5 | 281.8 ± 13 | 263.1 ± 8.1 | 0.979 | 0.192 |
Asp | 12.2 ± 1 | 14 ± 0.9 | 11.7 ± 0.8 | 11.5 ± 0.7 | 0.366 | 0.087 |
Glu | 50.2 ± 1.8 | 66.6 ± 3.5 ** | 52.8 ± 2.7 | 56.1 ± 1.5 | <0.001 | 0.124 |
Thr | 149.4 ± 6.5 | 175.1 ± 9.3 * | 186.2 ± 7.6 | 194.6 ± 6.9 | 0.03 | 0.001 |
Ala | 285.5 ± 5.5 | 344 ± 30.6 | 377.3 ± 21.7 | 321.7 ± 8.2 * | 0.942 | 0.102 |
Pro | 76.1 ± 4 | 96.7 ± 10.7 | 88.4 ± 3.2 | 93.7 ± 3.5 | 0.025 | 0.407 |
Lys | 266.5 ± 12.5 | 301.3 ± 15.6 | 348.4 ± 16.1 | 325 ± 8.4 | 0.673 | <0.001 |
Cys | 2.1 ± 0.3 | 1.7 ± 0.1 | 1.7 ± 0.1 | 1.9 ± 0.2 | 0.79 | 0.61 |
Tyr | 51.1 ± 2 | 79 ± 4.5 *** | 50.5 ± 2.2 | 64.2 ± 2 *** | <0.001 | 0.009 |
Met | 50.6 ± 1.9 | 63 ± 3.4 ** | 58 ± 1.5 | 57.3 ± 1.2 | 0.006 | 0.673 |
Val | 212.1 ± 6.8 | 198.2 ± 9.6 | 188.2 ± 9.5 | 209.5 ± 7.4 | 0.665 | 0.462 |
Ile | 96.3 ± 2.9 | 86.3 ± 5 | 83.9 ± 3.6 | 94.6 ± 2.6 | 0.927 | 0.593 |
Leu | 170.6 ± 6.5 | 148.9 ± 7.5 | 150.1 ± 7.7 | 159.7 ± 8.6 | 0.435 | 0.531 |
Phe | 78.2 ± 1.7 | 79 ± 3.3 | 72.7 ± 1.8 | 78.9 ± 2.9 | 0.173 | 0.263 |
Trp | 70 ± 3 | 90.2 ± 3.5 *** | 90.5 ± 4.6 | 89.3 ± 5.3 | 0.033 | 0.027 |
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He, Y.; Su, J.; Gao, H.; Li, J.; Feng, Z.; Yin, Y. Untargeted Metabolomics Reveals the Function of GPRC6A in Amino Acid and Lipid Metabolism in Mice. Metabolites 2022, 12, 776. https://doi.org/10.3390/metabo12090776
He Y, Su J, Gao H, Li J, Feng Z, Yin Y. Untargeted Metabolomics Reveals the Function of GPRC6A in Amino Acid and Lipid Metabolism in Mice. Metabolites. 2022; 12(9):776. https://doi.org/10.3390/metabo12090776
Chicago/Turabian StyleHe, Yumin, Jingyun Su, Hongrui Gao, Jianzhong Li, Zemeng Feng, and Yulong Yin. 2022. "Untargeted Metabolomics Reveals the Function of GPRC6A in Amino Acid and Lipid Metabolism in Mice" Metabolites 12, no. 9: 776. https://doi.org/10.3390/metabo12090776