Hostname: page-component-848d4c4894-p2v8j Total loading time: 0 Render date: 2024-06-12T07:24:25.893Z Has data issue: false hasContentIssue false
  • English
  • Français

Inhibition of FASN reduces the synthesis of medium-chain fatty acids in goat mammary gland

Published online by Cambridge University Press:  09 June 2014

J. J. Zhu ,
J. Luo ,
W. Wang ,
K. Yu ,
H. B. Wang ,
H. B. Shi ,
Y. T. Sun ,
X. Z. Lin  and
J. Li
J. J. Zhu
Affiliation:
Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, PR China
J. Luo*
Affiliation:
Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, PR China
W. Wang
Affiliation:
Department of Biological Sciences, University of Alabama, Tuscaloosa, USA
K. Yu
Affiliation:
Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, PR China
H. B. Wang
Affiliation:
Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, PR China
H. B. Shi
Affiliation:
Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, PR China
Y. T. Sun
Affiliation:
Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, PR China
X. Z. Lin
Affiliation:
Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, PR China
J. Li
Affiliation:
Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, PR China
*
†E-mail: luojun@nwsuaf.edu.cn
Get access

Abstract

Fatty acid synthase (FASN) is known as a crucial enzyme of cellular de novo fatty acid synthesis in mammary gland which has been proved as the main source of short and medium-chain fatty acids of milk. However, the regulatory role of FASN in goat-specific milk fatty acids composition remains unclear. We cloned and analyzed the full-length of FASN gene from the mammary gland of Capra hircus (Xinong Saanen dairy goat) (DQ 915966). Comparative gene expression analysis suggested that FASN is predominantly expressed in fat, small intestine and mammary gland tissues, and expresses higher level at lactation period. Inhibition of FASN activity by different concentrations (0, 5, 15, 25 and 35 μM) of orlistat, a natural inhibitor of FASN, resulted in decreased expression of acetyl-CoA carboxylase α (ACCα), lipoprotein lipase and heart-type fatty acid binding protein (H-FABP) in a concentration-dependent manner in goat mammary gland epithelial cells (GMEC). Similar results were also obtained by silencing of FASN. Additionally, reduction of FASN expression also led to apparent decline of the relative content of decanoic acid (C10:0) and lauric acid (C12:0) in GMEC. Our study provides a direct evidence for inhibition of FASN reduces cellular medium-chain fatty acids synthesis in GMEC.

Keywords

goatfatty acid synthaseorlistatshRNAfatty acids composition
Type
Research Article
Information
animal , Volume 8 , Issue 9 , September 2014 , pp. 1469 - 1478
Copyright
© The Animal Consortium 2014 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Amy, CM, Williams-Ahlf, B, Naggert, J and Smith, S 1992. Intron-exon organization of the gene for the multifunctional animal fatty acid synthase. Proceedings of the National Academy of Sciences of the United States of America 89, 1105–1108.CrossRefGoogle ScholarPubMed
Bernard, L, Leroux, C, Rouel, J, Bonnet, M and Chilliard, Y 2012. Effect of the level and type of starchy concentrate on tissue lipid metabolism, gene expression and milk fatty acid secretion in Alpine goats receiving a diet rich in sunflower-seed oil. British Journal of Nutrition 107, 1147–1159.CrossRefGoogle Scholar
Bionaz, M and Loor, JJ 2008. ACSL1, AGPAT6, FABP3, LPIN1, and SLC27A6 are the most abundant isoforms in bovine mammary tissue and their expression is affected by stage of lactation. Journal of Nutrition 138, 1019–1024.CrossRefGoogle ScholarPubMed
Carey, EM, Dils, R and Hanse, HJ 1970. Short communications. Chain-length specificity for termination of atty acid biosynthesis by fatty acid synthetase complexes from lactating rabbit mamary gland and rat liver. Biochemical Journal 117, 633–635.CrossRefGoogle ScholarPubMed
Chajès, V, Cambot, M, Moreau, K, Lenoir, GM and Joulin, V 2006. Acetyl-CoA carboxylase α is essential to breast cancer cell survival. Cancer Research 66, 5287–5294.CrossRefGoogle ScholarPubMed
Chawla, A, Boisvert, WA, Lee, CH, Laffitte, BA, Barak, Y, Joseph, SB, Liao, D, Nagy, L, Edwards, PA, Curtiss, LK, Evans, RM and Tontonoz, P 2001. A PPAR gamma-LXR-ABCA1 pathway in macrophages is involved in cholesterol efflux and atherogenesis. Molecular Cell 7, 161–171.CrossRefGoogle ScholarPubMed
Cho, KH, Kim, MJ, Jeon, GJ and Chung, HY 2011. Association of genetic variants for FABP3 gene with back fat thickness and intramuscular fat content in pig. Molecular Biology Reports 38, 2161–2166.CrossRefGoogle ScholarPubMed
Chuang, HY, Chang, YF and Hwang, JJ 2011. Antitumor effect of orlistat, a fatty acid synthase inhibitor, is via activation of caspase-3 on human colorectal carcinoma-bearing animal. Biomedicine & Pharmacotherapy 65, 286–292.CrossRefGoogle ScholarPubMed
Folch, J, Lees, M and Sloane Stanley, GH 1957. A simple method for the isolation and purification of total lipides from animal tissues. Journal of Biological Chemistry 226, 497–509.CrossRefGoogle ScholarPubMed
Foster, DW 2012. Malonyl-CoA: the regulator of fatty acid synthesis and oxidation. Journal of Clinical Investigation 122, 1958–1959.CrossRefGoogle ScholarPubMed
Green, CD, Ozguden-Akkoc, CG, Wang, Y, Jump, DB and Olson, LK 2010. Role of fatty acid elongases in determination of de novo synthesized monounsaturated fatty acid species. Journal of Lipid Research 51, 1871–1877.CrossRefGoogle ScholarPubMed
Grunnet, I and Knudsen, J 1978. Medium chain acyl-thioester hydrolase activity in goat and rabbit mammary gland fatty acid synthetase complexes. Biochemical and Biophysical Research Communications 80, 745–749.CrossRefGoogle ScholarPubMed
Ha, JK and Lindsay, RC 1993. Release of volatile branched-chain and other fatty acids from ruminant milk fats by various lipases. Journal of Dairy Science 76, 677–690.Google ScholarPubMed
Haenlein, GFW 2004. Goat milk in human nutrition. Small Ruminant Research 51, 155–163.CrossRefGoogle Scholar
Hansen, HJM, Carey, EM and Dils, R 1970. Fatty acid biosynthesis VII. Substrate control of chain-length of products synthesised by rat liver fatty acid synthetase. Biochimica et Biophysica Acta (BBA) – Lipids and Lipid Metabolism 210, 400–410.CrossRefGoogle ScholarPubMed
Harvatine, KJ, Boisclair, YR and Bauman, DE 2009. Recent advances in the regulation of milk fat synthesis. Animal 3, 40–54.CrossRefGoogle ScholarPubMed
Hillgartner, FB, Salati, LM and Goodridge, AG 1995. Physiological and molecular mechanisms involved in nutritional regulation of fatty acid synthesis. Physiological Reviews 75, 47–76.CrossRefGoogle ScholarPubMed
Hussein, M, Harvatine, KH, Weerasinghe, WMPB, Sinclair, LA and Bauman, DE 2013. Conjugated linoleic acid-induced milk fat depression in lactating ewes is accompanied by reduced expression of mammary genes involved in lipid synthesis. Journal of Dairy Science 96, 3825–3834.CrossRefGoogle ScholarPubMed
Jayakumar, A, Tai, MH, Huang, WY, al-Feel, W, Hsu, M, Abu-Elheiga, L, Chirala, SS and Wakil, SJ 1995. Human fatty acid synthase: properties and molecular cloning. Proceedings of the National Academy of Sciences of the United States of America 92, 8695–8699.CrossRefGoogle ScholarPubMed
Kameda, K and Goodridge, AG 1991. Isolation and partial characterization of the gene for goose fatty acid synthase. Journal of Biological Chemistry 266, 419–426.CrossRefGoogle ScholarPubMed
Knudsen, J and Grunnet, I 1980. Primer specificity of mammalian mammary gland fatty acid synthetases. Biochemical and Biophysical Research Communications 95, 1808–1814.CrossRefGoogle ScholarPubMed
Kusudo, T, Kontani, Y, Kataoka, N, Ando, F, Shimokata, H and Yamashita, H 2011. Fatty acid-binding protein 3 stimulates glucose uptake by facilitating AS160 phosphorylation in mouse muscle cells. Genes to Cells 16, 681–691.CrossRefGoogle ScholarPubMed
Lin, XZ, Luo, J, Zhang, LP, Wang, W, Shi, HB and Zhu, JJ 2013. miR-27a suppresses triglyceride accumulation and affects gene mRNA expression associated with fat metabolism in dairy goat mammary gland epithelial cells. Gene 521, 15–23.CrossRefGoogle ScholarPubMed
McGarry, JD, Mannaerts, GP and Foster, DW 1977. A possible role for malonyl-CoA in the regulation of hepatic fatty acid oxidation and ketogenesis. Journal of Clinical Investigation 60, 265–270.CrossRefGoogle ScholarPubMed
Mikkelsen, J, Højrup, P, Hansen, HF, Hansen, JK and Knudsen, J 1985. Evidence that the medium-chain acyltransferase of lactating-goat mammary-gland fatty acid synthetase is identical with the acetyl/malonyltransferase. Biochemical Journal 227, 981–985.CrossRefGoogle ScholarPubMed
Nomura, DK, Long, JZ, Niessen, S, Hoover, HS, Ng, SW and Cravatt, BF 2010. Monoacylglycerol lipase regulates a fatty acid network that promotes cancer pathogenesis. Cell 140, 49–61.CrossRefGoogle ScholarPubMed
Olsen, CP 2011. Why make fat?: the impact of de novo fattyacid synthesis on fat storage, reproduction, and longevity in c. e/egans. PhD thesis, University of Washington, Saint Louis.Google Scholar
Rangan, VS and Smith, S 1996. Expression in Escherichia coli and refolding of the malonyl-/acetyltransferase domain of the multifunctional animal fatty acid synthase. Journal of Biological Chemistry 271, 31749–31755.CrossRefGoogle ScholarPubMed
Roy, R, Taourit, S, Zaragoza, P, Eggen, A and Rodellar, C 2005. Genomic structure and alternative transcript of bovine fatty acid synthase gene (FASN): comparative analysis of the FASN gene between monogastric and ruminant species. Cytogenetic and Genome Research 111, 65–73.CrossRefGoogle ScholarPubMed
Rudolph, MC, McManaman, JL, Phang, T, Russell, T, Kominsky, DJ, Serkova, NJ, Stein, T, Anderson, SM and Neville, MC 2007. Metabolic regulation in the lactating mammary gland: a lipid synthesizing machine. Physiological Genomics 28, 323–336.CrossRefGoogle ScholarPubMed
Sankaranarayanapillai, M, Zhang, N, Baggerly, KA and Gelovani, JG 2012. Metabolic shifts induced by fatty acid synthase inhibitor orlistat in non-small cell lung carcinoma cells provide novel pharmacodynamic biomarkers for positron emission tomography and magnetic resonance spectroscopy. Molecular Imaging and Biology 15, 136–147.CrossRefGoogle Scholar
Schmid, B, Rippmann, JF, Tadayyon, M and Hamilton, BS 2005. Inhibition of fatty acid synthase prevents preadipocyte differentiation. Biochemical and Biophysical Research Communications 328, 1073–1082.CrossRefGoogle ScholarPubMed
Shi, H and Shen, L 2010. The alteration of extracellular signal-regulated kinase (ERK) activity of COS7 cells from interphase to mitosis. Cellular and Molecular Biology 56, OL1427–OL1433.Google Scholar
Shi, HB, Luo, J, Zhu, JJ, Li, J, Sun, YT, Lin, XZ, Zhang, LP, Yao, DW and Shi, HP 2013. PPARG regulates genes involved in triacylglycerol synthesis and secretion in mammary gland epithelial cells of dairy goats. PPAR Research 2013, 1–10.CrossRefGoogle Scholar
Smith, S 1994. The animal fatty acid synthase: one gene, one polypeptide, seven enzymes. FASEB Journal 8, 1248–1259.CrossRefGoogle ScholarPubMed
Thompson, BJ and Smith, S 1985. Biosynthesis of fatty acids by lactating human breast epithelial cells: an evaluation of the contribution to the overall composition of human milk fat. Pediatric Research 19, 139–143.CrossRefGoogle Scholar
Wang, W, Luo, J, Zhong, Y, Lin, XZ, Shi, HB, Zhu, JJ, Li, J, Sun, YT and Zhao, WS 2012. Goat liver X receptor α, molecular cloning, functional characterization and regulating fatty acid synthesis in epithelial cells of goat mammary glands. Gene 505, 114–120.CrossRefGoogle ScholarPubMed
Williams, C 2000. Dietary fatty acids and human health. Annales de Zootechnie 49, 165–180.CrossRefGoogle Scholar
Yang, WS, Nevin, DN, Peng, R, Brunzell, JD and Deeb, SS 1995. A mutation in the promoter of the lipoprotein lipase (LPL) gene in a patient with familial combined hyperlipidemia and low LPL activity. Proceedings of the National Academy of Sciences of the United States of America 92, 4462–4466.CrossRefGoogle Scholar
Supplementary material: PDF

Zhu Supplementary Material

Table S1 and Figures S1-S2

Download Zhu Supplementary Material(PDF)
PDF 236.3 KB