Hostname: page-component-8448b6f56d-tj2md Total loading time: 0 Render date: 2024-04-24T17:53:17.069Z Has data issue: false hasContentIssue false
  • English
  • Français

Molecular characterization of fibroblast growth factor-16 and its role in promoting the differentiation of intramuscular preadipocytes in goat

Published online by Cambridge University Press:  22 June 2020

K. Huang ,
J. J. Liang ,
Y. Q. Lin ,
J. J. Zhu ,
J. Q. Ma  and
Y. Wang
K. Huang
Affiliation:
College of Life Science and Technology, Southwest Minzu University, 610041, Chengdu, Sichuan, China Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, 610041, Chengdu, Sichuan, China
J. J. Liang
Affiliation:
College of Life Science and Technology, Southwest Minzu University, 610041, Chengdu, Sichuan, China Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, 610041, Chengdu, Sichuan, China
Y. Q. Lin
Affiliation:
College of Life Science and Technology, Southwest Minzu University, 610041, Chengdu, Sichuan, China Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, 610041, Chengdu, Sichuan, China
J. J. Zhu
Affiliation:
College of Life Science and Technology, Southwest Minzu University, 610041, Chengdu, Sichuan, China Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, 610041, Chengdu, Sichuan, China Key Laboratory of Sichuan Province for Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Exploitation, Southwest Minzu University, 610041, Chengdu, Sichuan, China
J. Q. Ma
Affiliation:
College of Life Science and Technology, Southwest Minzu University, 610041, Chengdu, Sichuan, China Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, 610041, Chengdu, Sichuan, China
Y. Wang*
Affiliation:
College of Life Science and Technology, Southwest Minzu University, 610041, Chengdu, Sichuan, China Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, 610041, Chengdu, Sichuan, China Key Laboratory of Sichuan Province for Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Exploitation, Southwest Minzu University, 610041, Chengdu, Sichuan, China
*
†E-mail: wangyong010101@hotmail.com
Get access

Abstract

Fat metabolism is an important and complex biochemical reaction in vivo and is regulated by many factors. Recently, the findings on high expression of fibroblast growth factor-16 (FGF16) in brown adipose tissue have led to an interest in exploring its role in lipogenesis and lipid metabolism. The study cloned the goat’s FGF16 gene 624 bp long, including the complete open reading frame that encodes 207 amino acids. We found that FGF16 expression is highest in goat kidneys and hearts, followed by subcutaneous fat and triceps. Moreover, the expression of FGF16 reached its peak on the 2nd day of adipocyte differentiation (P < 0.01) and then decreased significantly. We used overexpression and interference to study the function of FGF16 gene in goat intramuscular preadipocytes. Silencing of FGF16 decreased adipocytes lipid droplet aggregation and triglyceride synthesis. This is in contrast to the situation where FGF16 is overexpressed. Furthermore, knockdown of FGF16 also caused down-regulated expression of genes associated with adipocyte differentiation including CCAAT enhancer-binding protein beta (P < 0.01), fatty acid-binding protein-2 (P < 0.01) and sterol regulatory element binding protein-1 (P < 0.05), but the preadipocyte factor-1 was up-regulated. At the same time, the genes adipose triglyceride lipase (P < 0.01) and hormone-sensitive lipase (P < 0.05) associated with triglyceride breakdown were highly expressed. Next, we locked the fibroblast growth factor receptor-4 (FGFR4) through the protein interaction network and interfering with FGF16 to significantly reduce FGFR4 expression. It was found that the expression profile of FGFR4 in adipocyte differentiation was highly similar to that of FGF16. Overexpression and interference methods confirmed that FGFR4 and FGF16 have the same promoting function in adipocyte differentiation. Finally, using co-transfection technology, pc-FGF16 and siRNA-FGFR4, siRNA2-FGF16 and siRNA-FGFR4 were combined to treat adipocytes separately. It was found that in the case of overexpression of FGF16, cell lipid secretion and triglyceride synthesis showed a trend of first increase and then decrease with increasing interference concentration. In the case of interference with FGF16, lipid secretion and triglyceride synthesis showed a downward trend with the increase of interference concentration. These findings illustrated that FGF16 mediates adipocyte differentiation via receptor FGFR4 expression and contributed to further study of the functional role of FGF16 in goat fat formation.

Keywords

adipocyteclonefibroblast growth factor receptor-4regulationtriglyceride
Type
Research Article
Information
animal , Volume 14 , Issue 11 , November 2020 , pp. 2351 - 2362
Copyright
© The Author(s), 2020. Published by Cambridge University Press on behalf of The Animal Consortium

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

Antoine, M, Wirz, W, Tag, CG, Gressner, AM, Wycislo, M, Müller, R and Kiefer, P 2006. Fibroblast growth factor 16 and 18 are expressed in human cardiovascular tissues and induce on endothelial cells migration but not proliferation. Biochemical and Biophysical. Research Communications 346, 224–233.CrossRefGoogle Scholar
Beenken, A and Mohammadi, M 2009. The FGF family: biology, pathophysiology and therapy. Nature Reviews Drug Discovery 8, 235–253.CrossRefGoogle ScholarPubMed
Basu, M, Mukhopadhyay, S, Chatterjee, U and Roy, SS 2014. FGF16 promotes invasive behavior of SKOV-3 ovarian cancer cells through activation of mitogen-activated protein kinase (MAPK) signaling pathway. The Journal of Biological Chemistry 289, 1415–1428.CrossRefGoogle ScholarPubMed
Cristancho, AG and Lazar, MA 2011. Forming functional fat: a growing understanding of adipocyte differentiation. Nature Reviews Molecular Cell Biology 12, 722–734.CrossRefGoogle ScholarPubMed
Chapman, SC, Cai, Q, Bleyl, SB and Schoenwolf, GC 2006. Restricted expression of Fgf16 within the developing chick inner ear. Developmental Dynamics 235, 2276–2281.Google ScholarPubMed
Deng, KP, Ren, CF, Liu, ZF, Gao, XX, Fan, YX, Zhang, GM, Zhang, YL, Ei-Samahy, MA, Wang, F and You, PH 2018. Characterization of RUNX1T1, an Adipogenesis Regulator in Ovine Preadipocyte Differentiation. International Journal of Molecular Sciences 19, 1300.CrossRefGoogle ScholarPubMed
Eseberri, I, Miranda, J, Lasa, A, Mosqueda-Solís, A, González-Manzano, S, Santos-Buelga, C and Portillo, MP 2019. Effects of quercetin metabolites on triglyceride metabolism of 3T3-L1 preadipocytes and mature adipocytes. International Journal of Molecular Sciences 20, 264.CrossRefGoogle ScholarPubMed
Grimaldi, PA 2001. The roles of PPARs in adipocyte differentiation. Progress in Lipid Research 40, 269–281.CrossRefGoogle ScholarPubMed
Hotta, Y, Sasaki, S, Konishi, M, Kinoshita, H, Kuwahara, K, Nakao, K and Itoh, N 2008. Fgf16 Is required for cardiomyocyte proliferation in the mouse embryonic heart. Developmental Dynamics: An Official Publication of the American Association of Anatomists 237, 2947–2954.Google ScholarPubMed
Jing, R, Feng, H, Jiang, N, Zhang, H, Fang, W, Ni, Z and Yuan, J 2019. Visceral adipogenesis inhibited by Pref-1 is associated with peritoneal angiogenesis. Nephrology (Carlton, Vic.). 25, 248–254.Google ScholarPubMed
Kim, EJ, Kang, MJ, Seo, YB, Nam, SW and Kim, GD 2018. Acer okamotoanum nakai leaf extract inhibits adipogenesis via suppressing expression of PPAR γ and C/EBP α in 3T3-L1 cells. Journal of Microbiology and Biotechnology 28, 1645–1653.CrossRefGoogle Scholar
Konishi, M, Mikami, T, Yamasaki, M, Miyake, A and Itoh, N 2000. Fibroblast growth factor-16 is a growth factor for embryonic brown adipocytes. The Journal of Biological Chemistry 275, 12119–12122.CrossRefGoogle ScholarPubMed
Litthauer, D and Serrero, G 1992. The primary culture of mouse adipocyte precursor cells in defined medium. Comparative Biochemistry and Physiology. A, Comparative Physiology 101, 59–64.CrossRefGoogle ScholarPubMed
Luo, X, Jia, R, Li, K, Zhu, X, Zhao, D, Whitehead, JP and Yan, J 2015. Fibroblast growth factor-1 inhibits Wnt/β-catenin pathway during adipogenesis. Journal of Central South University. Medical Sciences 40, 843–850.Google ScholarPubMed
Lu, SY, Sheikh, F, Sheppard, PC, Fresnoza, A, Duckworth, ML, Detillieux, KA and Cattini, PA 2008. FGF-16 is required for embryonic heart development. Biochemical and Biophysical Research Communications 373, 270–274.CrossRefGoogle ScholarPubMed
Miyake, A, Chitose, T, Kamei, E, Murakami, A, Nakayama, Y, Konishi, M and Itoh, N 2014. Fgf16 is required for specification of GABAergic neurons and oligodendrocytes in the zebrafish forebrain. PLoS ONE 9, e110836.Google ScholarPubMed
Mota de Sá, P, Richard, AJ, Hang, H and Stephens, JM 2017. Transcriptional regulation of adipogenesis. Comprehensive Physiology 7, 635–674.Google ScholarPubMed
MacDougald, OA and Lane, MD 1995. Transcriptional regulation of gene expression during adipocyte differentiation. Annual Review of Biochemistry 64, 345–373.CrossRefGoogle ScholarPubMed
McPherson, R and Gauthier, A 2004. Molecular regulation of SREBP function: the Insig-SCAP connection and isoform-specific modulation of lipid synthesis. Biochemistry and Cell Biology 82, 201–211.CrossRefGoogle ScholarPubMed
Navre, M and Ringold, GM 1989. Differential effects of fibroblast growth factor and tumor promoters on the initiation and maintenance of adipocyte differentiation. The Journal of Cell Biology 109, 1857–1863.Google ScholarPubMed
Nomura, R, Kamei, E, Hotta, Y, Konishi, M, Miyake, A and Itoh, N 2006. Fgf16 is essential for pectoral fin bud formation in zebrafish. Biochemical and Biophysical Research Communications 347, 340–346.Google ScholarPubMed
Ornitz, DM and Itoh, N 2015. The fibroblast growth factor signaling pathway. Wiley Interdisciplinary Reviews. Developmental Biology 4, 215–266.CrossRefGoogle ScholarPubMed
Olaya-Sánchez, D, Sánchez-Guardado, LÓ, Ohta, S, Chapman, SC, Schoenwolf, GC, Puelles, L and Hidalgo-Sánchez, M 2017. Fgf3 and Fgf16 expression patterns in the developing chick inner ear. Brain Structure and Function 222, 131–149.Google ScholarPubMed
Rulifson, IC, Collins, P, Miao, L, Nojima, D, Lee, KJ, Hardy, M, Gupte, J, Hensley, K, Samayoa, K, Cam, C, Rottman, JB, Ollmann, M, Richards, WG and Li, Y 2017. In vitro and in vivo analyses reveal profound effects of fibroblast growth factor 16 as a metabolic regulator. The Journal of Biological Chemistry 292, 1951–1969.CrossRefGoogle ScholarPubMed
Rodbell, M 1964. Metabolism of isolated fat cells. I. Effects of hormones on glucose metabolism and lipolysis. The Journal of Biological Chemistry 239, 375–380.Google ScholarPubMed
Seo, HA and Lee, IK 2013. The role of Nrf2: adipocyte differentiation, obesity, and insulin resistance. Oxidative Medicine and Cellular Longevity 2013, 1–7.CrossRefGoogle Scholar
Sun, Y, Wang, R, Zhao, S, Li, W, Liu, W, Tang, L, Wang, Z, Wang, W, Liu, R, Ning, G, Wang, J and Hong, J 2019. FGF9 inhibits browning program of white adipocytes and associates with human obesity. Journal of Molecular Endocrinology 62, 79–90.CrossRefGoogle ScholarPubMed
Shang, ZC, Guo, L, Wang, N, Shi, H, Wang, YX and Li, H 2014. Oleate promotes differentiation of chicken primary preadipocytes in vitro. Bioscience Reports 34, e00093.Google ScholarPubMed
Wang, J, Jin, Y and Cattini, PA 2017. Expression of the cardiac maintenance and survival factor FGF-16 gene Is regulated by Csx/Nkx2.5 and is an early target of doxorubicin cardiotoxicity. DNA and Cell Biology 36, 117–126.CrossRefGoogle ScholarPubMed
Xie, T and Leung, PS 2017. Fibroblast growth factor 21: a regulator of metabolic disease and health span. American Journal of Physiology. Endocrinology and Metabolism 313, E292–E302.CrossRefGoogle ScholarPubMed
Xu, Q, Lin, S, Zhu, J and Lin, YQ 2018a. The expression stability analysis of reference genes in the process of goat intramuscular preadipocytes differentiation. Acta Veterinaria et Zootechnica Sinica 49, 907–918.Google Scholar
Xu, Q, Wang, Y, Zhu, J, Zhao, Y and Lin, YQ 2018b. Molecular characterization of GTP binding protein overexpressed in skeletal muscle (GEM) and its role in promoting adipogenesis in goat intramuscular preadipocytes. Animal Biotechnology 20, 1–8.Google Scholar
Xiong, Y, Xu, Q, Lin, S, Wang, Y, Lin, YQ and Zhu, JJ 2018. Knockdown of LXRα Inhibits Goat Intramuscular Preadipocyte Differentiation. International Journal of Molecular Sciences 19, 3037.CrossRefGoogle ScholarPubMed
Zhang, X, Ibrahimi, OA, Olsen, SK, Umemori, H, Mohammadi, M and Ornitz, DM 2006. Receptor specificity of the fibroblast growth factor family. The complete mammalian FGF family. The Journal of Biological Chemistry 281, 15694–15700.Google ScholarPubMed