Effect of dietary Sophora alopecuroides supplementation on differential expression of intramuscular fat-related genes of Ningxia Tan sheep as determined by transcriptome sequencing


Xin G.S., Guo J., Yu B.J., Li H., Gu Y.L., Gao Q.X., Zhang J. (2022): Effect of dietary Sophora alopecuroides supplementation on differential expression of intramuscular fat-related genes of Ningxia Tan sheep as determined by transcriptome sequencing. Czech J. Anim. Sci., 67: 185–197.

supplementary materialdownload PDF

The present study investigated the effect of different dietary levels (0%, 0.25%, 0.5%, 1%, 2%) of Sophora alopecuroides (S. alopecuroides) on intramuscular fat (IMF) content in Ningxia Tan sheep. Candidate genes affecting IMF deposition were screened by comparing the RNA-Seq profiles of test (2% S. alopecuroides) and control (no S. alopecuroides) groups. The results showed that the intramuscular fat content of the diet supplemented with S. alopecuroides was significantly lower than that of the control group, and the intramuscular fat content decreased significantly with the increase of the dietary content of S. alopecuroides (< 0.05). The highest IMF content of the control group (group A) was 4.21%, which was significantly higher than that of the other four experimental groups (B, C, D, E) (< 0.05). Moreover, 66 genes were significantly upregulated and 202 genes were significantly downregulated in the test group compared to the control. Gene Ontology functional annotation revealed that the differentially expressed genes were closely related to plasma lipoprotein particle assembly, plasma lipoprotein particle organization, protein-lipid complex subunit organization, and other biological processes. KEGG pathway analysis showed that the differentially expressed genes belonged to 147 metabolic pathways. Finally, four differential genes which may be related to IMF deposition were screened out in PPAR signalling pathway, fatty acid biosynthesis, fatty acid degradation, and fatty acid metabolism. The quantitative analysis of four differential genes including ACSL3, PLIN2, ABCA1 and ANGPTL4 was carried out by RT-qPCR method, and the results were basically consistent with those of RNA-seq. Among them, the quantitative results of PLIN2 gene were slightly different from the sequencing results, but the difference was not significant, and they may be false positive. This study lays the foundation for understanding the molecular mechanism regulating mutton quality, and provides a theoretical basis for the study of S. alopecuroides as a feed additive to improve mutton meat quality and impact the gene function.

Aronsson L, Huang Y, Parini P, Korach-Andre M, Hakansson EM, Gustafsson JA, Pettersson S, Arulampalam V, Rafter J. Decreased fat storage by lactobacillus paracasei is associated with increased levels of angiopoietin-like 4 protein (ANGPTL4). PLoS One. 2010 Sep 30;5(9): 7 p.  https://doi.org/10.1371/journal.pone.0013087
Brooks-Wilson A, Marcil M, Clee SM, Zhang LH, Roomp K, Dam M, Yu L, Brewer C, Collins JA, Molhuizen HO, Loubser O. Mutations in ABC1 in Tangier disease and familial high-density lipoprotein deficiency. Nat Genet. 1999 Aug;22:336-45.  https://doi.org/10.1038/11905
Carr RM, Peralta G, Yin XY, Ahima RS. Absence of perilipin 2 prevents hepatic steatosis, glucose intolerance and ceramide accumulation in alcohol-fed mice. PLoS One. 2014 May 15;9(5): 10 p.  https://doi.org/10.1371/journal.pone.0097118
Chen G, Sui Y. Production, performance, slaughter characteristics, and meat quality of Ziwuling wild crossbred pigs. Trop Anim Health Pro. 2018 Feb;50(2):365-72. https://doi.org/10.1007/s11250-017-1441-2
Chen Y, Qiao Y, Xiao Y, Chen H, Zhao L, Huang M, Zhou G. Differences in physicochemical and nutritional properties of breast and thigh meat from crossbred chickens, commercial broilers, and spent hens. Asian-Australas J Anim. 2016 Jun;29(6):855-64. https://doi.org/10.5713/ajas.15.0840
Chen J, Chen F, Lin X, Wang Y, He J, Zhao Y. Effect of excessive or restrictive energy on growth performance, meat quality, and intramuscular fat deposition in finishing Ningxiang Pigs. Animals. 2021 Dec 25;11(1): 14 p. https://doi.org/10.3390/ani11010027
Davoli R, Gandolfi G, Braglia S, Comella M, Zambonelli P, Buttazzoni L, Russo V. New SNP of the porcine perilipin 2 (PLIN2) gene, association with carcass traits and expression analysis in skeletal muscle. Mol Biol Rep. 2011 Mar;38(3):1575-83.  https://doi.org/10.1007/s11033-010-0266-0
Eck F, Phuyal S, Smith MD, Kaulich M, Wilkinson S, Farhan H, Behrends C. ACSL3 is a novel GABARAPL2 interactor that links ufmylation and lipid droplet biogenesis. J Cell Sci. 2020 Sep 16;133(18): 38 p. https://doi.org/10.1242/jcs.243477
Feng SQ, Chen XD, Xia T, Gan L, Qiu H, Dai MH, Zhou L, Peng Y, Yang ZQ. Cloning, chromosome mapping and expression characteristics of porcine ANGPTL3 and-4. Cytogenet Genome Res. 2006 Jun;114(6):44-9.  https://doi.org/10.1159/000091927
Forde N, Mehta JP, McGettigan PA, Mamo S, Bazer FW, Spencer TE, Lonergan P. Alterations in expression of endometrial genes coding for proteins secreted into the uterine lumen during conceptus elongation in cattle. BMC Genom. 2013 May 10;14(1): 13 p. https://doi.org/10.1186/1471-2164-14-321
Gandolfi G, Mazzoni M, Zambonelli P, Lalatta-Costerbosa G, Tronca A, Russo V, Davoli R. Perilipin 1 and perilipin 2 protein localization and gene expression study in skeletal muscles of European cross-breed pigs with different intramuscular fat contents. Meat Sci. 2011 Aug;88(4):631-7. https://doi.org/10.1016/j.meatsci.2011.02.020
Guo Q, Kong XF, Hu CJ, Zhou B, Wang CT, Shen QW. Fatty acid content, flavor compounds, and sensory quality of pork loin as affected by dietary supplementation with L-arginine and glutamic acid. J Food Quality. 2019 Nov;84(12):3445-53.  https://doi.org/10.1111/1750-3841.14959
Hocquette JF, Cassar-Malek I, Jurie C, Bauchart D, Picard B, Renand G. Relationships between muscle growth potential, intramuscular fat content and different indicators of muscle fibre types in young Charolais bulls. Anim Sci J. 2012 Nov;83(11):750-8. https://doi.org/10.1111/j.1740-0929.2012.01021.x
Hu YW, Ma X, Huang JL, Mao XR, Yang JY, Zhao JY, Li SF, Qiu YR, Yang J, Zheng L, Wang Q. Dihydrocapsaicin attenuates plaque formation through a PPARγ/LXRα pathway in apoE−/− mice fed a high-fat/high-cholesterol diet. PLoS One. 2013 Jun 26;8(6): 13 p.  https://doi.org/10.1371/journal.pone.0066876
Legry V, Bokor S, Cottel D, Beghin L, Catasta G, Nagy E, Gonzalez-Gross M, Spinneker A, Stehle P, Molnar D, Moreno LA. Associations between common genetic polymorphisms in angiopoietin-like proteins 3 and 4 and lipid metabolism and adiposity in european adolescents and adults. J Clin Endocr Metab. 2009 Nov;94(12):5070-7.  https://doi.org/10.1210/jc.2009-0769
Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014 Dec;15(12): 21 p.  https://doi.org/10.1186/s13059-014-0550-8
Lv YT, Guan WT, Qiao HZ, Wang CX, Chen F, Zhang YZ, Liao ZC. Veterinary medicine and omics (veterinomics): Metabolic transition of milk triacylglycerol synthesis in sows from late pregnancy to lactation. OMICS J Integr Biol. 2015 Oct 1;19(10):602-16. https://doi.org/10.1089/omi.2015.0102
Lv Y, Cao Y, Gao Y, Yun JY, Yu YS, Zhang LC, Hu Z, Liu L, Xue J, Zhang. Effect of ACSL3 expression levels on preadipocyte differentiation in Chinese Red Steppe Cattle. DNA Cell Biol. 2019 Sep 10;38(9):945-54.  https://doi.org/10.1089/dna.2018.4443
Mamedova LK, Robbins K, Johnson BJ, Bradford BJ. Tissue expression of angiopoietin-like protein 4 in cattle. J Anim Sci. 2010 Jan 1;88(1):124-30.  https://doi.org/10.2527/jas.2009-2258
NRC – National Research Council. Nutrient requirements of small ruminants. 1st ed. Washington, DC, USA: National Academies Press; 2007. 142 p.
Olfaz M, Ocak N, Erener G, Cam MA, Garipoglu AV. Growth, carcass and meat characteristics of Karayaka growing rams fed sugar beet pulp, partially substituting for grass hay as forage. Meat Sci. 2005 May 1;70(1):7-14. https://doi.org/10.1016/j.meatsci.2004.11.015
Owens NDL, De Domenico E, Gilchrist MJ. An RNA-Seq protocol for differential expression analysis. Cold Spring Harb Protoc. 2019 Jun 1;2019(6): 10 p. https://doi.org/10.1101/pdb.prot098368
Poppelreuther M, Rudolph B, Du C, Grossmann R, Becker M, Thiele C, Ehehalt R, Fullekrug J. The N-terminal region of acyl-CoA synthetase 3 is essential for both the localization on lipid droplets and the function in fatty acid uptake. J Lipid Res. 2012 May 1;53(5):888-900. https://doi.org/10.1194/jlr.M024562
Ren ZQ, Wu WJ, Liu WH, Zheng R, Li JL, Zuo B, Xu DQ, Li FE, Lei MG, Ni DB, Xiong YZ. Differential expression and effect of the porcine ANGPTL4 gene on intramuscular fat. Genet Mol Res. 2014 Apr 16;13(2):2949-58. https://doi.org/10.4238/2014.April.16.3
Ruth MH, Jean M, Chris M, Anne MM, Torres S, Anita T, Michael TC, Grace CD. Functional analysis of muscle gene expression profiles associated with tenderness and intramuscular fat content in pork. Meat Sci. 2012 Dec;92(4):440-50. https://doi.org/10.1016/j.meatsci.2012.05.007
Sastre D, Costa NN, Sa ALA, Conceicao SDB, Chiaratti MR, Adona PR, Guemra S, Meirelles FV, Santos SD, Sena L, Ohashi OM. Expression of PLIN2 and PLIN3 during oocyte maturation and early embryo development in cattle. Theriogenology. 2014 Jan 15;81(2):326-31.  https://doi.org/10.1016/j.theriogenology.2013.10.002
Suzuki K, Takahashi K, Nishimaki-Mogami T, Kagechika H, Yamamoto M, Itabe H. Docosahexaenoic acid induces adipose differentiation-related protein through activation of retinoid X receptor in human choriocarcinoma BeWo cells. Biol Pharm Bull. 2009 Jul 1;32(7):1177-82. https://doi.org/10.1248/bpb.32.1177
Tang YY, Chen WJ, Lu Q, Tang CK. Research advances of intracellular transport and function of ABCA1. Prog Biochem Biophys. 2013 Jun 1;40(6):510-9. https://doi.org/10.3724/SP.J.1206.2012.00419
Wang Z, Gerstein M, Snyder M. RNA-Seq: A revolutionary tool for transcriptomics. Nat Rev Genet. 2009 Jan;10(1):57-63.  https://doi.org/10.1038/nrg2484
Yao MH, Ma RL, Li SG, Mu LT, Ding YS, Guo SX. ABCA1 gene polymorphisms and the interactions with obesity risk factors in patients with low HDL-C. Acta Nutrimen Sin. 2016 May;38(5):443-7.
Zhang JH, Wu Y, Hou JH, Ma QZ. Determination of the contents of free amio acids in Sophora alopecuroides. Amino Acids Biotic Resour. 1997 Apr;19(4):39-40.
supplementary materialdownload PDF

© 2022 Czech Academy of Agricultural Sciences | Prohlášení o přístupnosti