Hostname: page-component-cd9895bd7-dk4vv Total loading time: 0 Render date: 2024-12-27T03:35:19.178Z Has data issue: false hasContentIssue false

Negative effect of insulin-induced gene 2 on milk fat synthesis in buffalo mammary epithelial cells

Published online by Cambridge University Press:  19 January 2022

Xinyang Fan
Affiliation:
Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming650201, Yunnan, China
Yongyun Zhang
Affiliation:
Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming650201, Yunnan, China Teaching Demonstration Center of the Basic Experiments of Agricultural Majors, Yunnan Agricultural University, Kunming650201, Yunnan, China
Lihua Qiu
Affiliation:
Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming650201, Yunnan, China
Yongwang Miao*
Affiliation:
Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming650201, Yunnan, China
*
Author for correspondence: Yongwang Miao, Email: [email protected]

Abstract

Insulin-induced gene 2 (INSIG2) is a recently identified gene that is implicated in the regulation of cholesterol metabolism and lipogenesis in mammals. Although the data in goats emphasizes a role for INSIG2 in milk fat synthesis, the regulatory mechanism in buffalo is not clear. In this study, we analyzed the protein abundance of INSIG2 at peak lactation and dry-off period in buffalo mammary tissue. The results indicated that, relative to the peak lactation, the protein abundance of INSIG2 in the dry-off period was higher. To determine the function of INSIG2 in milk fat synthesis, INSIG2 was overexpressed and knocked down by lentiviral transfection in buffalo mammary epithelial cells (BuMECs). The response to overexpressing INSIG2 included down-regulation of SREBP, PPARG, FASN, ELOVL6, SCD, APGAT6 and TIP47 coupled with a decrease in content of triacylglycerol (TAG). However, in response to knockdown of INSIG2, the significant increase in content of TAG along with marked up-regulation of SREBP, PPARG, FASN, ELOVL6, SCD, APGAT6 and TIP47 suggests that INSIG2 negatively affects milk fat synthesis in BuMECs. No significant difference in mRNA abundance of GPAM and DGAT2 in response to overexpression or interference of INSIG2 indicates that they might also be influenced by other regulatory factors. Taken together, our results provide strong support for the negative effect of INSIG2 on milk fat synthesis in BuMECs.

Type
Research Article
Copyright
Copyright © The Author(s), 2022. 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, 35193529.CrossRefGoogle ScholarPubMed
Bernard, L, Toral, PG and Chilliard, Y (2017) Comparison of mammary lipid metabolism in dairy cows and goats fed diets supplemented with starch, plant oil, or fish oil. Journal of Dairy Science 100, 93389351.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
Chen, C, Hsu, L, Huang, K, Goto, S, Chen, C and Nakano, T (2017) Overexpression of insig-2 inhibits atypical antipsychotic-induced adipogenic differentiation and lipid biosynthesis in adipose-derived stem cells. Scientific Reports 7, 10901.CrossRefGoogle ScholarPubMed
Deng, T, Pang, C, Ma, X, Lu, X, Duan, A, Zhu, P and Liang, X (2016) Four novel polymorphisms of buffalo INSIG2 gene are associated with milk production traits in Chinese buffaloes. Molecular and Cellular Probes 30, 294299.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, 24892498.CrossRefGoogle ScholarPubMed
Fan, X, Qiu, L, Teng, X, Zhang, Y and Miao, Y (2020) Effect of INSIG1 on the milk fat synthesis of buffalo mammary epithelial cells. Journal of Dairy Research 87, 349355.CrossRefGoogle ScholarPubMed
Farr, VC, Stelwagen, K, Cate, LR, Molenaar, AJ, Mcfadden, TB and Davis, SR (1996) An improved method for the routine biopsy of bovine mammary tissue. Journal of Dairy Science 79, 543549.CrossRefGoogle 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, 4054.CrossRefGoogle ScholarPubMed
Jo, Y, Cha, J and Moon, Y (2017) Regulation of INSIG2 by microRNA-96. Animal Cells and Systems 21, 263268.CrossRefGoogle 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. Journal of Clinical Endocrinology and Metabolism 93, 19952001.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, 17361746.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, 21482156.CrossRefGoogle ScholarPubMed
Radhakrishnan, A, Ikeda, Y, Kwon, HJ, Brown, MS and Goldstein, JL (2007) Sterol-regulated transport of SREBPs from endoplasmic reticulum to Golgi: oxysterols block transport by binding to Insig. Academy of Sciences of the United States of America 104, 65116518.Google ScholarPubMed
Ren, C, Wang, L, Fan, Y, Jia, R, Zhang, G, Deng, M, Deng, K and Wang, F (2017) Scd1 contributes to lipid droplets formation in GMEC via transcriptional regulation of Tip47 and Adrp. European Journal of Lipid Science and Technology 120, 1700238. doi.org/10.1002/ejlt.201700238.CrossRefGoogle Scholar
Rincon, G, Islas-Trejo, A, Castillo, AR, Bauman, DE, German, BJ and Medrano, JF (2012) Polymorphisms in genes in the SREBP1 signalling pathway and SCD are associated with milk fatty acid composition in Holstein cattle. Journal of Dairy Research 79, 6675.CrossRefGoogle ScholarPubMed
Sato, R (2010) Sterol metabolism and SREBP activation. Archives of Biochemistry and Biophysics 501, 177181.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, 78447853.CrossRefGoogle ScholarPubMed
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, 49874995.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 of the United States of America 101, 71067111.CrossRefGoogle ScholarPubMed
Tumanovskaa, LV, Swansonb, RJ, Serebrovskaa, ZO, Portnichenkoa, GV, Goncharova, SV, Kysilova, BA, Moibenkoa, OO and Dosenko, VE (2019) Cholesterol enriched diet suppresses ATF6 and PERK and upregulates the IRE1 pathways of the unfolded protein response in spontaneously hypertensive rats: relevance to pathophysiology of atherosclerosis in the setting of hypertension. Pathophysiology 26, 219226.CrossRefGoogle Scholar
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, 3341.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, 783795.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, 3172631734.CrossRefGoogle ScholarPubMed
Supplementary material: PDF

Fan et al. supplementary material

Fan et al. supplementary material

Download Fan et al. supplementary material(PDF)
PDF 530.9 KB