Abstract
The preadipocytes differentiation is a vital process of lipogenesis; exploring the molecular mechanisms of lipogenesis contributes to improve the meat quality and final commercial income. Lipogenesis has been widely reported in other livestock, but little is known about the gene expression profiles at different stages during preadipocytes differentiation in sheep. In this study, ovine preadipocytes were cultured in vitro and then induced to begin differentiation. Then, the gene expression profiles of preadipocytes collected on day 0 (D0), day 2 (D2), and day 8 (D8) of differentiation were analyzed by RNA-seq technology. According to the findings, 2254 differentially expressed genes (DEGs) were found in D2 vs D0; 1817 DEGs and 1902 DEGs were found in D8 vs D0 and D8 vs D2, respectively. The DEGs were found to be enriched in several biological processes, including focal adhesion, ECM-receptor interaction, PI3K-Akt signaling pathway, steroid biosynthesis, and MAPK signaling pathway, according to Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. The regulatory network of the DEGs related to ovine preadipocytes differentiation was systematically constructed, which showed that hub genes might modulate ovine preadipocytes differentiation. In summary, preadipocyte differentiation is regulated by several key genes, including ACACB, CXCL6, SREBF1, INSIG1, APOE, GJA1, CDH11, SYNE1, PCSK1, S100A4, FN1, PLIN2, CXCL6, FN1, PTX3, and FABP3. This study provides a deeper knowledge of the roles of genes in sheep lipogenesis by revealing global gene expression profiles during preadipocyte differentiation.
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References
Abdalla BA, Chen J, Nie Q, Zhang X (2018) Genomic insights into the multiple factors controlling abdominal fat deposition in a chicken model. Front Genet 9:262. https://doi.org/10.3389/fgene.2018.00262
Ai Hasan M, Martin PE, Shu X, Patterson S, Bartholomew C (2021) Type III collagen is required for adipogenesis and actin stress fibre formation in 3T3-L1 preadipocytes. Biomolecules 11:156. https://doi.org/10.3390/biom11020156
Ayala-Sumuano JT, Velez-Delvalle C, Beltrán-Langarica A, Marsch-Moreno M, Cerbón-Solorzano J, Kuri-Harcuch W (2011) Srebf1a is a key regulator of transcriptional control for adipogenesis. Sci Rep 1:178. https://doi.org/10.1038/srep00178
Bernasochi GB, Bell JR, Simpson ER, Delbridge L, Boon WC (2019) Impact of estrogens on the regulation of white, beige, and brown adipose tissue depots. Compr Physiol 9:457–475. https://doi.org/10.1002/cphy.c180009
Bonacina F, Moregola A, Porte R, Baragetti A, Bonavita E, Salatin A, Grigore L, Pellegatta F, Molgora M, Sironi M, Barbati E, Mantovani A, Bottazzi B, Catapano AL, Garlanda C, Norata GD (2019) Pentraxin 3 deficiency protects from the metabolic inflammation associated to diet-induced obesity. Cardiovasc Res 115:1861–1872. https://doi.org/10.1093/cvr/cvz068
Buras ED, Converso-Baran K, Davis CS, Akama T, Hikage F, Michele DE, Brooks SV, Chun TH (2019) Fibro-adipogenic remodeling of the diaphragm in obesity-associated respiratory dysfunction. Diabetes 68:45–56. https://doi.org/10.2337/db18-0209
Chen S, Zhou Y, Chen Y, Gu J (2018) Fastp: an ultra-fast all-in-one FASTQ preprocessor. Bioinformatics 34:884–890. https://doi.org/10.1093/bioinformatics/bty560
Cornelius P, MacDougald OA, Lane MD (1994) Regulation of adipocyte development. Annu Rev Nutr 14:99–129. https://doi.org/10.1146/annurev.nu.14.070194.000531
Côté JA, Guénard F, Lessard J, Lapointe M, Biron S, Vohl MC, Tchernof A (2017) Temporal changes in gene expression profile during mature adipocyte dedifferentiation. Int J Genomics 5149362. https://doi.org/10.1155/2017/5149362
Cristancho AG, Lazar MA (2011) Forming functional fat: a growing understanding of adipocyte differentiation. Nat Rev Mol Cell Biol 12:722–734. https://doi.org/10.1038/nrm3198
Deng K, Ren C, Fan Y, Liu Z, Zhang G, Zhang Y, You P, Wang F (2020) miR-27a is an important adipogenesis regulator associated with differential lipid accumulation between intramuscular and subcutaneous adipose tissues of sheep. Domest Anim Endocrinol 71:106393. https://doi.org/10.1016/j.domaniend.2019.106393
Dong XY, Tang SQ (2010) Insulin-induced gene: a new regulator in lipid metabolism. Peptides 31:2145–2150. https://doi.org/10.1016/j.peptides.2010.07.020
Ernst J, Bar-Joseph Z (2006) STEM: a tool for the analysis of short time series gene expression data. BMC Bioinformatics 7:191. https://doi.org/10.1186/1471-2105-7-191
Fan X, Qiu L, Teng X, Zhang Y, Miao Y (2020) Effect of INSIG1 on the milk fat synthesis of buffalo mammary epithelial cells. J Dairy Res 87:349–355. https://doi.org/10.1017/S0022029920000710
Gregoire FM, Smas CM, Sul HS (1998) Understanding adipocyte differentiation. Physiol Rev 78:783–809. https://doi.org/10.1152/physrev.1998.78.3.783
Guo H, Qiu X, Deis J, Lin TY, Chen X (2020) Pentraxin 3 deficiency exacerbates lipopolysaccharide-induced inflammation in adipose tissue. Int J Obes (lond) 44:525–538. https://doi.org/10.1038/s41366-019-0402-4
Hou S, Jiao Y, Yuan Q, Zhai J, Tian T, Sun K, Chen Z, Wu Z, Zhang J (2018) S100A4 protects mice from high-fat diet-induced obesity and inflammation. Lab Invest 98:1025–1038. https://doi.org/10.1038/s41374-018-0067-y
Ibrahim MM (2010) Subcutaneous and visceral adipose tissue: structural and functional differences. Obes Rev 11:11–18. https://doi.org/10.1111/j.1467-789X.2009.00623.x
Ignacio RM, Gibbs CR, Lee ES, Son DS (2016) Differential chemokine signature between human preadipocytes and adipocytes. Immune Netw 16:189–194. https://doi.org/10.4110/in.2016.16.3.189
Jin X, Hao Z, Zhao M, Shen J, Ke N, Song Y, Qiao L, Lu Y, Hu L, Wu X, Wang J, Luo Y (2021a) MicroRNA-148a regulates the proliferation and differentiation of ovine preadipocytes by targeting PTEN. Animals 11:820. https://doi.org/10.3390/ani11030820
Jin X, Wang J, Hu J, Liu X, Li S, Lu Y, Zhen H, Li M, Zhao Z, Luo Y (2021b) MicroRNA-200b regulates the proliferation and differentiation of ovine preadipocytes by targeting p27 and KLF9. Animals 11:2417. https://doi.org/10.3390/ani11082417
Kanehisa M, Araki M, Goto S, Hattori M, Hirakawa M, Itoh M, Katayama T, Kawashima S, Okuda S, Tokimatsu T, Yamanishi Y (2008) KEGG for linking genomes to life and the environment. Nucleic Acids Res 36:480–484. https://doi.org/10.1093/nar/gkm882
Kang D, Zhou G, Zhou S, Zeng J, Wang X, Jiang Y, Yang Y, Chen Y (2017) Comparative transcriptome analysis reveals potentially novel roles of Homeobox genes in adipose deposition in fat-tailed sheep. Sci Rep 7:14491. https://doi.org/10.1038/s41598-017-14967-9
Kim S, Lee N, Park ES, Yun H, Ha TU, Jeon H, Yu J, Choi S, Shin B, Yu J, Rhee SD, Choi Y, Rho J (2021) T-cell death associated gene 51 is a novel negative regulator of PPARγ that inhibits PPARγ-RXRα heterodimer formation in adipogenesis. Mol Cells. 44:1–12. https://doi.org/10.14348/molcells.2020.0143
Krautgasser C, Mandl M, Hatzmann FM, Waldegger P, Mattesich M, Zwerschke W (2019) Reliable reference genes for expression analysis of proliferating and adipogenically differentiating human adipose stromal cells. Cell Mol Biol Lett 24:14–25. https://doi.org/10.1186/s11658-019-0140-6
Lasrich D, Bartelt A, Grewal T, Heeren J (2015) Apolipoprotein E promotes lipid accumulation and differentiation in human adipocytes. Exp Cell Res 337:94–102. https://doi.org/10.1016/j.yexcr.2015.07.015
Li H, Ma Z, Jia L, Li Y, Xu C, Wang T, Han R, Jiang R, Li Z, Sun G, Kang X, Liu X (2016) Systematic analysis of the regulatory functions of microRNAs in chicken hepatic lipid metabolism. Sci Rep 6:31766. https://doi.org/10.1038/srep31766
Ma YN, Wang B, Wang ZX, Gomez NA, Zhu MJ, Du M (2018) Three-dimensional spheroid culture of adipose stromal vascular cells for studying adipogenesis in beef cattle. Animal 22:1–7. https://doi.org/10.1017/S1751731118000150
Miao X, Luo Q, Qin X, Guo Y, Zhao H (2015) Genome-wide mRNA-seq profiling reveals predominant down-regulation of lipid metabolic processes in adipose tissues of Small Tail Han than Dorset sheep. Biochem Biophys Res Commun 467:413–420. https://doi.org/10.1016/j.bbrc.2015.09.129
Mo D, Yu K, Chen H, Chen L, Liu X, He Z, Cong P, Chen Y (2017) Transcriptome landscape of porcine intramuscular adipocytes during differentiation. J Agric Food Chem 65:6317–6328. https://doi.org/10.1021/acs.jafc.7b02039
Nematbakhsh S, Chong PP, Selamat J, Nordin N, Idris LH, AbdullRazis AF (2021) Molecular regulation of lipogenesis, adipogenesis and fat deposition in chicken. Genes 12:414. https://doi.org/10.3390/genes12030414
Shao W, Espenshade PJ (2012) Expanding roles for SREBP in metabolism. Cell Metab 16:414–419. https://doi.org/10.1016/j.cmet.2012.09.002
Shimba S, Wada T, Hara S, Tezuka M (2004) EPAS1 promotes adipose differentiation in 3T3-L1 cells. J Biol Chem 279:40946–40953. https://doi.org/10.1074/jbc.M400840200
Sun MY, Wang SJ, Li XQ, Shen YL, Lu JR, Tian XH, Rahman K, Zhang LJ, Nian H, Zhang H (2019) CXCL6 Promotes renal interstitial fibrosis in diabetic nephropathy by activating JAK/STAT3 signaling pathway. Front Pharmacol 10:224. https://doi.org/10.3389/fphar.2019.00224
Tian Y, Zhao Y, Yu W, Melak S, Niu Y, Wei W, Zhang L, Chen J (2022) ACAT2 is a novel negative regulator of pig intramuscular preadipocytes differentiation. Biomolecules 12:237. https://doi.org/10.3390/biom12020237
Trapnell C, Williams BA, Pertea G, Mortazavi A, Kwan G, van Baren MJ, Salzberg SL, Wold BJ, Pachter L (2010) Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation. Nat Biotechnol 28:511–515. https://doi.org/10.1038/nbt.1621
Wang Y, Fu Y, Yan Z, Zhang XB, Pei M (2019) Impact of fibronectin knockout on proliferation and differentiation of human infrapatellar fat pad-derived stem cells. Front Bioeng Biotechnol 7:321. https://doi.org/10.3389/fbioe.2019.00321
Wu Y, Chen K, Li L, Hao Z, Wang T, Liu Y, Xing G, Liu Z, Li H, Yuan H, Lu J, Zhang C, Zhang J, Zhao D, Wang J, Nie J, Ye D, Pan G, Chan WY, Liu X (2022) Plin2-mediated lipid droplet mobilization accelerates exit from pluripotency by lipidomic remodeling and histone acetylation. Cell Death Differ 29:2316–2331. https://doi.org/10.1038/s41418-022-01018-8
Xiao C, Wei T, Liu LX, Liu JQ, Wang CX, Yuan ZY, Ma HH, Jin HG, Zhang LC, Cao Y (2021) Whole-transcriptome analysis of preadipocyte and adipocyte and construction of regulatory networks to investigate lipid metabolism in sheep. Front Genet 12:662143. https://doi.org/10.3389/fgene.2021.662143
Zhang T, Zhang X, Han K, Zhang G, Wang J, Xie K, Xue Q (2017) Genome-Wide Analysis of lncRNA and mRNA Expression during Differentiation of Abdominal Preadipocytes in the Chicken. G3 7:953–966. https://doi.org/10.1534/g3.116.037069
Zhang M, Li F, Ma XF, Li WT, Jiang RR, Han RL, Li GX, Wang YB, Li ZY, Tian YD, Kang XT, Sun GR (2019) Identification of differentially expressed genes and pathways between intramuscular and abdominal fat-derived preadipocyte differentiation of chickens in vitro. BMC Genomics 20:743. https://doi.org/10.1186/s12864-019-6116-0
Zhou G, Wang X, Yuan C, Kang D, Xu X, Zhou J, Geng R, Yang Y, Yang Z, Chen Y (2017) Integrating miRNA and mRNA expression profiling uncovers miRNAs underlying fat deposition in sheep. Biomed Res Int 1–11. https://doi.org/10.1155/2017/1857580
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This study was financially supported by the Youth Fund of College of Animal Science and Technology of Gansu Agricultural University (GAU-DK-QNJJ-202102).
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ZH, XJ, and JW conceived and designed the experiments; ZH, YL, JH, XL, and SL conducted the experiments; ZH, ML, and FZ analyzed the data; ZH and JW drafted and revised the manuscript. All authors read and approved the final manuscript.
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All animal procedures in this study were approved by Animal Experiment Ethics Committee of Gansu Agricultural University with an approval number of GSAU-ETH-AST-2021–027.
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Hao, Z., Jin, X., Wang, J. et al. Functional differentiation of the ovine preadipocytes —insights from gene expression profiling. Funct Integr Genomics 23, 97 (2023). https://doi.org/10.1007/s10142-023-01034-y
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DOI: https://doi.org/10.1007/s10142-023-01034-y