Hostname: page-component-8448b6f56d-dnltx Total loading time: 0 Render date: 2024-04-24T02:10:24.928Z Has data issue: false hasContentIssue false
  • English
  • Français

Effect of INSIG1 on the milk fat synthesis of buffalo mammary epithelial cells

Published online by Cambridge University Press:  10 September 2020

Xinyang Fan ,
Lihua Qiu ,
Xiaohong Teng ,
Yongyun Zhang  and
Yongwang Miao
Xinyang Fan
Affiliation:
Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, Yunnan, 650201, China
Lihua Qiu
Affiliation:
Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, Yunnan, 650201, China
Xiaohong Teng
Affiliation:
Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, Yunnan, 650201, China
Yongyun Zhang
Affiliation:
Teaching Demonstration Center of the Basic Experiments of Agricultural Majors, Yunnan Agricultural University, Kunming, Yunnan, 650201, China
Yongwang Miao*
Affiliation:
Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, Yunnan, 650201, China
*
Author for correspondence: Yongwang Miao, Email: yongwangmiao1@126.com
Get access Rights & Permissions [Opens in a new window]

Abstract

We hypothesized that insulin-induced gene 1 (INSIG1) affects milk fat synthesis in buffalo. For this reason, the protein abundance of INSIG1 in the mammary tissue of buffalo during the peak period of lactation and dry-off period was evaluated. The results showed that the expression of INSIG1 at the peak of lactation was lower than that in the dry-off period. To explore the role of INSIG1 in milk fat synthesis, the buffalo mammary epithelial cells (BMECs) were isolated and purified from buffalo mammary tissue, and INSIG1 gene were overexpressed and knocked down by constructing the recombinant lentivirus vector of INSIG1 gene and transfecting into BMECs. Results revealed that INSIG1 overexpression decreased the expression of INSIG2, SREBP, PPARG, SCD, GPAM, DGAT2 and AGPAT6, which led to reduction of triglycerides (TAG) content in the cell. In contrast, knockdown of INSIG1 had a positive effect on mRNA expression of the above genes. Overall, the data provide strong support for a key role of INSIG1 in the regulation of milk fat synthesis in BMECs.

Keywords

BuffaloINSIG1lentivirusmilk fat synthesis
Type
Research Article
Information
Journal of Dairy Research , Volume 87 , Issue 3 , August 2020 , pp. 349 - 355
Copyright
Copyright © The Author(s), 2020. Published by Cambridge University Press on behalf of Hannah Dairy Research Foundation.

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.)

Footnotes

*

These authors contributed equally to this paper.

References

Abeni, F, Degano, L, Calza, F, Giangiacomo, R and Pirlo, G (2005) Milk quality and automatic milking: fat globule size, natural creaming, and lipolysis. Journal of Dairy Science 88, 3519–3529.CrossRefGoogle ScholarPubMed
Attie, AD (2004) Insig: a significant integrator of nutrient and hormonal signals. Journal of Clinical Investigation 113, 1112–1114.CrossRefGoogle ScholarPubMed
Bionaz, M and Loor, JJ (2008) Gene networks driving bovine milk fat synthesis during the lactation cycle. BMC Genomics 9, 366.CrossRefGoogle ScholarPubMed
Carobbio, S, Hagen, RM, Lelliott, CJ, Slawik, M, Medina-Gomez, G, Tan, C, Sicard, A, Atherton, HJ, Barbarroja, N, Bjursell, M, Bohlooly-Y, M, Virtue, S, Tuthill, A, Lefai, E, Laville, M, Wu, T, Considine, RV, Vidal, H, Langin, D, Oresic, M, Tinahones, FJ, Fernandez-Real, JM, Griffin, JL, Seth, JK, López, M and Vidal-Puig, A (2013) Adaptive changes of the Insig1/SREBP1/SCD1 set point help adipose tissue to cope With increased storage demands of obesity. Diabetes 62, 3697–3708.CrossRefGoogle ScholarPubMed
Diamond, RH, Du, K, Lee, VM, Mohn, KL, Haber, BA, Tewari, DS and Taub, R (1993) Novel delayed-early and highly insulin-induced growth response genes. Identification of HRS, a potential regulator of alternative pre-mRNA splicing. Journal of Biological Chemistry 268, 15185–15192.Google ScholarPubMed
Dong, XY and Tang, SQ (2010) Insulin-induced gene: a new regulator in lipid metabolism. Peptides 31, 2145–2150.CrossRefGoogle ScholarPubMed
Engelking, LJ, Kuriyama, H, Hammer, RE, Horton, JD, Brown, MS, Goldstein, JL and Liang, G (2004) Overexpression of Insig-1 in the livers of transgenic mice inhibits SREBP processing and reduces insulin-stimulated lipogenesis. Journal of Clinical Investigation 113, 1168–1175.CrossRefGoogle ScholarPubMed
Engelking, LJ, Liang, G, Hammer, RE, Takaishi, K, Kuriyama, H, Evers, BM, Li, W, Horton, JD, Goldstein, JL and Brown, MS (2005) Schoenheimer effect explained – feedback regulation of cholesterol synthesis in mice mediated by Insig proteins. Journal of Clinical Investigation 115, 2489–2498.Google ScholarPubMed
Espenshade, PJ and Hughes, AL (2007) Regulation of sterol synthesis in Eukaryotes. Annual Review of Genetics 41, 401–427.CrossRefGoogle ScholarPubMed
Feramisco, JD, Goldstein, JL and Brown, MS (2004) Membrane topology of human Insig-1, a protein regulator of lipid synthesis. Journal of Biological Chemistry 279, 8487–8496.CrossRefGoogle ScholarPubMed
Gong, Y, Lee, JN, Lee, PCW, Goldstein, JL, Brown, MS and Ye, J (2006) Sterol-regulated ubiquitination and degradation of Insig-1 creates a convergent mechanism for feedback control of cholesterol synthesis and uptake. Cell Metabolism 3, 15–24.CrossRefGoogle ScholarPubMed
Han, LQ, Li, HJ, Wang, YY, Zhu, HS, Wang, LF, Guo, YJ, Lu, WF, Wang, YL and Yang, GY (2010) mRNA abundance and expression of SLC27A, ACC, SCD, FADS, LPIN, INSIG, and PPARGC1 gene isoforms in mouse mammary glands during the lactation cycle. Genetics and Molecular Research: GMR 9, 1250–1257.Google ScholarPubMed
Harvatine, KJ, Boisclair, YR and Bauman, DE (2009) Recent advances in the regulation of milk fat synthesis. Animal: An International Journal of Animal Bioscience 3, 40–54.CrossRefGoogle ScholarPubMed
Horton, JD, Goldstein, JL and Brown, MS (2002) SREBPs: activators of the complete program of cholesterol and fatty acid synthesis in the liver. Journal of Clinical Investigation 109, 1125–1131.CrossRefGoogle ScholarPubMed
Horton, JD, Shah, NA, Warrington, JA, Anderson, NN, Park, SW, Brown, MS and Goldstein, JL (2003) Combined analysis of oligonucleotide microarray data from transgenic and knockout mice identifies direct SREBP target genes. Proceedings of the National Academy of Sciences 100, 12027–12032.CrossRefGoogle ScholarPubMed
Hu, H, Wang, J, Bu, D, Wei, H, Zhou, L, Li, F and Loor, JJ (2009) In vitro culture and characterization of a mammary epithelial cell line from Chinese Holstein dairy cow. PLoS ONE 4, e7636.CrossRefGoogle ScholarPubMed
Kast-Woelbern, HR, Dana, SL, Cesario, RM, Sun, L, de Grandpre, LY, Brooks, ME, Osburn, DL, Reifel-Miller, A, Klausing, K and Leibowitz, MD (2004) Rosiglitazone induction of insig-1 in white adipose tissue reveals a novel interplay of peroxisome proliferator-activated receptor gamma and sterol regulatory element-binding protein in the regulation of adipogenesis. Journal of Biological Chemistry 279, 23908–23915.Google ScholarPubMed
Krapivner, S, Popov, S, Chernogubova, E, Hellénius, M, Fisher, RM, Hamsten, A and Van'T Hooft, FM (2008) Insulin-induced gene 2 involvement in human adipocyte metabolism and body weight regulation. The Journal of Clinical Endocrinology and Metabolism 93, 1995–2001.CrossRefGoogle ScholarPubMed
Lee, JN, Gong, Y, Zhang, X and Ye, J (2006) Proteasomal degradation of ubiquitinated Insig proteins is determined by serine residues flanking ubiquitinated lysines. Proceedings of the National Academy of Sciences 103, 4958–4963.CrossRefGoogle ScholarPubMed
Leichner, GS, Avner, R, Harats, D and Roitelman, J (2009) Dislocation of HMG-CoA reductase and Insig-1, two polytopic endoplasmic reticulum proteins, en route to proteasomal degradation. Molecular Biology of the Cell 20, 3330–3341.CrossRefGoogle ScholarPubMed
Li, C, Wang, M, Zhang, T, He, Q, Shi, H, Luo, J and Loor, JJ (2019) Insulin-induced gene 1 and 2 isoforms synergistically regulate triacylglycerol accumulation, lipid droplet formation, and lipogenic gene expression in goat mammary epithelial cells. Journal of Dairy Science 102, 1736–1746.CrossRefGoogle ScholarPubMed
Liou, Y, Bai, YM, Lin, E, Chen, J, Chen, T, Hong, C and Tsai, S (2012) Gene-gene interactions of the INSIG1 and INSIG2 in metabolic syndrome in schizophrenic patients treated with atypical antipsychotics. The Pharmacogenomics Journal 12, 54–61.CrossRefGoogle ScholarPubMed
McFarlane, MR, Liang, G and Engelking, LJ (2014) Insig proteins mediate feedback inhibition of cholesterol synthesis in the intestine. Journal of Biological Chemistry 289, 2148–2156.CrossRefGoogle ScholarPubMed
Osorio, JS, Lohakare, J and Biona, M (2016) Biosynthesis of milk fat, protein, and lactose: roles of transcriptional and posttranscriptional regulation. Physiological Genomics 48, 231–256.CrossRefGoogle ScholarPubMed
Owen, JL, Zhang, Y, Bae, SH, Farooqi, MS, Liang, G, Hammer, RE, Goldstein, JL and Brown, MS (2012) Insulin stimulation of SREBP-1c processing in transgenic rat hepatocytes requires p70 S6-kinase. Proceedings of the National Academy of Sciences 109, 16184–16189.CrossRefGoogle ScholarPubMed
Shi, HB, Luo, J, Yao, DW, Zhu, JJ, Xu, HF, Shi, HP and Loor, JJ (2013) Peroxisome proliferator-activated receptor-γ stimulates the synthesis of monounsaturated fatty acids in dairy goat mammary epithelial cells Via The control of stearoyl-coenzyme A desaturase. Journal of Dairy Science 96, 7844–7853.CrossRefGoogle ScholarPubMed
Shi, H, Shi, H, Luo, J, Wang, W, Haile, AB, Xu, H and Li, J (2014) Establishment and characterization of a dairy goat mammary epithelial cell line with human telomerase (hT-MECs). Animal Science Journal 85, 735–743.CrossRefGoogle Scholar
Shi, HB, Wu, M, Zhu, JJ, Zhang, CH, Yao, DW, Luo, J and Loor, JJ (2017) Fatty acid elongase 6 plays a role in the synthesis of long-chain fatty acids in goat mammary epithelial cells. Journal of Dairy Science 100, 4987–4995.CrossRefGoogle Scholar
Song, S, Ou-Yang, Y, Huo, J, Zhang, Y, Yu, C, Liu, M, Teng, X and Miao, Y (2016) Molecular cloning, sequence characterization, and tissue expression analysis of three water buffalo (Bubalus Bubalis) genes – ST6GAL1, ST8SIA4, and SLC35C1. Archives Animal Breeding 59, 363–372.CrossRefGoogle Scholar
Takaishi, K, Duplomb, L, Wang, M, Li, J and Unger, RH (2004) Hepatic insig-1 or −2 overexpression reduces lipogenesis in obese Zucker diabetic fatty rats and in fasted/refed normal rats. Proceedings of the National Academy of Sciences 101, 7106–7111.CrossRefGoogle ScholarPubMed
Tsushima, H, Tang, YJ, Puviindran, V, Hsu, SC, Nadesan, P, Yu, C, Zhang, H, Mirando, AJ, Hilton, MJ and Alman, BA (2018) Intracellular biosynthesis of lipids and cholesterol by Scap and Insig in mesenchymal cells regulates long bone growth and chondrocyte homeostasis. Development (Cambridge, England) 145, v162396.CrossRefGoogle ScholarPubMed
Wu, C, Liu, L, Huo, J, Li, D, Yuan, Y, Yuan, F and Miao, Y (2014) Isolation, sequence characterization, and tissue transcription profiles of two novel buffalo genes: INSIG1 and INSIG2. Tropical Animal Health and Production 46, 33–41.CrossRefGoogle ScholarPubMed
Xu, HF, Luo, J, Zhao, WS, Yang, YC, Tian, HB, Shi, HB and Bionaz, M (2016) Overexpression of SREBP1 (sterol regulatory element binding protein 1) promotes de novo fatty acid synthesis and triacylglycerol accumulation in goat mammary epithelial cells. Journal of Dairy Science 99, 783–795.CrossRefGoogle ScholarPubMed
Yellaturu, CR, Deng, X, Park, EA, Raghow, R and Elam, MB (2009) Insulin enhances the biogenesis of nuclear sterol regulatory element-binding protein (SREBP)-1c by posttranscriptional down-regulation of Insig-2A and its dissociation from SREBP cleavage-activating protein (SCAP)⋅SREBP-1c complex. Journal of Biological Chemistry 284, 31726–31734.CrossRefGoogle ScholarPubMed
Zhou, L, Lin, Q, Wang, P, Yao, L, Leong, K, Tan, Z and Huang, Z (2017) Enhanced neuroprotective efficacy of bone marrow mesenchymal stem cells co-overexpressing BDNF and VEGF in a rat model of cardiac arrest-induced global cerebral ischemia. Cell Death and Disease 8, e2774.CrossRefGoogle Scholar
Zu, X, Yan, R, Robbins, S, Krishack, PA, Liao, DF and Cao, D (2007) Reduced 293T cell susceptibility to acrolein due to aldose reductase-like-1 protein expression. Toxicological Sciences 97, 562–568.CrossRefGoogle ScholarPubMed
Supplementary material: PDF

Fan et al. supplementary material

Tables S1-S3 and Figures S1-S3

Download Fan et al. supplementary material(PDF)
PDF 422.1 KB