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Exploration of Key Genes Affecting Intramuscular Fat Deposition in Guizhou Yellow Chicken (Gallus gallus domesticus) Based on RNA-seq Technology |
WANG Xiao-Ya1,2,3,4, CHEN Da-Hai1,2,3,4, RAO Yong-Chao1,2,3, YANG De-Feng1,2,3, TANG Hong1,2,3, LIN Jia-Dong1,2,3,4, ZHANG Fu-Ping1,2,3,4,* |
1 Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Ministry of Education, Guizhou University, Guiyang 550025, China; 2 Guizhou Provincial Key Laboratory of Animal Genetics, Breeding and Reproduction, Guiyang 550025, China; 3 College of Animal Science, Guizhou University, Guiyang 550025, China; 4 Research Chicken Farm of Guizhou University Guiyang 550025, China |
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Abstract Intramuscular fat (IMF) content was one of the important indicators to measure meat quality. However, the deposition of intramuscular fat content was affected by heredity, environment and nutrition level, and the deposition of intramuscular fat content was different among different varieties and strains, even among different individuals of the same strain. The aim of this study was to explore the differences in IMF content among different individuals of Guizhou yellow chickens (Gallus gallus domesticus) and to analyze the molecular regulation mechanism of IMF deposition in muscle of Guizhou yellow chickens by RNA-seq technique. 30 Guizhou yellow chickens hens were selected and raised in the same conditions to 120 d of age, and the IMF content in breast muscle was determined one by one after slaughter. There were 2 groups according to the level of IMF content: The high-IMF group (H group, n=4) and the low-IMF group (L group, n=4). In this study, RNA-seq technique was used to screen differentially expressed genes affecting IMF deposition in breast muscle of Guizhou yellow chicken; It also analyzed the differentially expressed genes for GO functional annotation and KEGG pathway enrichment. The results showed that a total of 259 differentially expressed genes were screened out from breast muscle tissues of individuals with high and low IMF content, including 64 up-regulated genes and 195 down-regulated genes. 6 genes were randomly selected for qPCR validation, and the qPCR results were consistent with the sequencing results, proved that the sequencing results were reliable. KEGG signaling pathway analysis showed that 5 signaling pathways, including focal adhesion, extracellular matrix (ECM) -receptor interaction, regulation of actin cytoskeleton, cell adhesion molecules, and TGF (transforming growth factor)-β signaling pathway, these pathways were significantly enriched. Calproteinase 2 (CAPN2), collagen type Ⅰ alpha 1 chain (COL1A1), COL1A2, collagen type Ⅵ alpha 1 chain (COL6A1), COL6A2, COL6A3, lipin1 (LPIN1) and phospholipid transfer protein (PLTP) were found in adhesion spots, ECM-receptor interaction, TGF-beta signaling pathway and PPAR signaling pathway. These genes may be key genes affecting IMF deposition in Guizhou yellow chickens. The differential expression genes identified in this study can be used as key genes for IMF deposition and provide a reference to explore the molecular regulation mechanism of IMF deposition in poultry in future.
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Received: 01 July 2021
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Corresponding Authors:
*zfu-1010@126.com
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[1] 段安琴, 陆杏蓉, 马小娅, 等. 2020. LPIN1基因对水牛乳腺上皮细胞甘油三酯和脂肪酸合成的影响[J].中国畜牧兽医, 47(10): 3140-3148. (DUAN A Q, LU X R, MA X Y, et al.2020. The effect of LPIN1 gene on triglyceride and fatty acid synthesis of bovine mammary epithelial cells[J]. Chinese animal husbandry and veterinary medicine, 47(10): 3140-3148.) [2] 李优磊. 2018. PPAR信号通路在调控猪皮下脂肪与肌内脂肪差异沉积中的作用及机制研究[D]. 博士学位论文, 西北农林科技大学,导师: 杨公社, pp. 20-30. (Li Y L.2018. Study on the role and mechanism of PPAR signaling pathway in regulating differential deposition of subcutaneous fat and intramuscular fat in pigs[D]. Thesis for Ph.D., Northwest Agriculture&Forestry University, Supervisor: Yang G S, pp. 20-30.) [3] Andraski A B.2019. Effects of replacing dietary monounsaturated fat with carbohydrate on HDL (High-Density Lipoprotein) protein metabolism and proteome composition in humans[J]. Arterioscler Thromb Vasc Biological, 39(11): 2411-2430. [4] Brooks M A, Choi C W, Lunt D K, et al.2011. Subcutaneous and intramuscular adipose tissue stearoyl-coenzyme A desaturase gene expression and fatty acid composition in calf- and yearling-fed Angus steers[J]. Journal of Animal Science, 89(8): 2556-2570. [5] Camou J P, Marchello J A, Thompson V F, et al.2007. Effect of postmortem storage on activity of mu- and m-calpain in five bovine muscles[J]. Journal of Animal Science, 85(10):2670-81. [6] Cánovas A, Quintanilla R, Amills M, et al.2010. Muscle transcriptomic profiles in pigs with divergent phenotypes for fatness traits[J]. BMC Genomics, 11(1): 1-15. [7] Carlson K B, Prusa K J, Fedler C A, et al.2017. Postmortem protein degradation is a key contributor to fresh pork loin tenderness[J]. Journal of Animal Science, 95(4):1574-1586. [8] Chabault M, E Baéza, Gigaud V, et al.2012. Analysis of a slow-growing line reveals wide genetic variability of carcass and meat quality-related traits[J]. BMC Genetics, 13(1): 90-90. [9] Chao X, Guo L, Wang Q, et al.2020. miR-429-3p/LPIN1 axis promotes chicken abdominal fat deposition via PPARγ pathway[J]. Frontiers in Cell and Developmental Biology, 8: 595637. [10] Cui H X, Zheng M Q, Liu R R, et al.2012. Liver dominant expression of fatty acid synthase (FAS) gene in two chicken breeds during intramuscular-fat development[J]. Molecular Biology Reports, 39(4): 3479-3484. [11] Du M, Yin J, Zhu M J.2010. Cellular signaling pathways regulating the initial stage of adipogenesis and marbling of skeletal muscle[J]. Meat Science, 86(1): 103-109. [12] Eaton J M, Mullins G R, Brindley D N, et al.2013. Phosphorylation of Lipin 1 and charge on the phosphatidic acid head group control its phosphatidic acid phosphatase activity and membrane association[J].The Journal of Biological Chemistry, 288(14): 9933-9945. [13] Hillege M, Caro R, Offringa C, et al.2020. TGF-β regulates collagen type I expression in myoblasts and myotubes via transient Ctgf and Fgf-2 expression[J]. Cells, 9(2):375. [14] Hocquette J F, Gondret F, Baeza E, et al.2010. Intramuscular fat content in meat-producing animals: Development, genetic and nutritional control, and identification of putative markers[J]. Animal, 4(2): 303-319. [15] Jiang H, Yazdanyar A, Lou B, et al.2015. Adipocyte phospholipid transfer protein and lipoprotein metabolism[J]. Arterioscler Thromb Vasc Biological, 35(2):316-322. [16] Khan M I, Jo C, Tariq M R.2015. Meat flavor precursors and factors influencing flavor precursors a systematic review[J]. Meat Science, 110: 278-284. [17] Li B, Weng Q, Dong C, et al.2018. A key gene, PLIN1, can affect porcine intramuscular fat content based on transcriptome analysis[J]. Genes, 9(4): 194. [18] Li D L, Chen J L, Wen J, et al.2013. Growth, carcase and meat traits and gene expression in chickens divergently selected for intramuscular fat content[J]. Poultry Science, 54(2): 183-189. [19] Li J, Xing S, Zhao G, et al.2020. Identification of diverse cell populations in skeletal muscles and biomarkers for intramuscular fat of chicken by single-cell RNA sequencing[J]. BMC Genomics, 21(1): 1-11. [20] Liu R, Zheng M, Wang J, et al.2019. Effects of genomic selection for intramuscular fat content in breast muscle in Chinese local chickens[J]. Animal Genetics, 50(1): 87-91. [21] Matitaputty P R, Wijaya C H, Bansi H, et al.2015. Influence of duck species and cross-breeding on sensory and quality characteristics of Alabio and Cihateup duck meat[J]. CyTA-Journal of Food, 13(4): 522-526. [22] Nzirorera, Carine, D' Souza, et al.2016. Lipid metabolism and signaling in cardiac lipotoxicity[J]. Biochimica Et Biophysica Acta Molecular & Cell Biology of Lipids, 1861(10): 1513-1524. [23] Ye M, Xu M, Chen C, et al., et al.2018. Expression analyses of candidate genes related to meat quality traits in squabs from two breeds of meat-type pigeon[J]. Journal of Animal Physiology and Animal Nutrition, 102(3): 727-735. [24] Mortimer S I, Werf V D,et al.Jacob D.L.2014. Genetic parameters for meat quality traits of australian lamb meat[J]. Meat Science, 96: 1016-1024. [25] Oh J, Kim C S, Min K, et al.2021. Type VI collagen and its cleavage product, endotrophin, cooperatively regulate the adipogenic and lipolytic capacity of adipocytes[J]. Metabolism, 114: 154430. [26] Pan X, Chen Z, Huang R, et al.2013. Transforming growth factor β1 induces the expression of collagen type I by DNA methylation in cardiac fibroblasts[J]. PLOS ONE, 8(4): e60335. [27] Peterson T R, Sengupta S S, Harris T E, et al.2011. mTOR complex 1 regulates lipin 1 localization to control the SREBP pathway[J]. Cell, 146: 408-20. [28] Sellers R S, Mahmood S R, Perumal G S, et al.2018. Phenotypic modulation of skeletal muscle fibers in lpin1-deficient lipodystrophic(fld) mice[J]. Veterinary Pathology, 56(2): 322-331. [29] Shu Z J, Hong K W, Tong X S, et al.2013. Transcriptome comparison between porcine subcutaneous and intramuscular stromal vascular cells during adipogenic differentiation[J]. PLOS ONE, 8(10): e77094. [30] Sponton CH, Hosono T, Taura J, et al.2020. The regulation of glucose and lipid homeostasis via PLTP as a mediator of BAT-liver communication[J]. EMBO Reports, 21(9): e49828. [31] Tous N, Lizardo R, Vila B, et al.2013. Effect of a high dose of cla in finishing pig diets on fat deposition and fatty acid composition in intramuscular fat and other fat depots[J]. Meat Science, 93(3): 517-524. [32] Tyra M, Ropka-Molik K, Terman A, et al.2013. Association between subcutaneous and intramuscular fat content in porcine ham and loin depending on age, breed and FABP3 and LEPR genes transcript abundance[J]. Molecular Biology Reports, 40(3): 2301-2308. [33] Zhang M, Li D, Zhai Y, et al.2020. The landscape of DNA methylation associated with the transcriptomic network of intramuscular adipocytes generates insight into intramuscular fat deposition in chicken[J]. Frontiers in Cell and Developmental Biology, 4(2): 8-206. [34] Zhang Y, Zan L, Wang H.2011. Screening candidate genes related to tenderness trait in Qinchuan cattle by genome array[J]. Molecular Biology Reports, 38(3): 2007-2014. |
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