Animal Bioscience

  • 제38권8호
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  • Pages.1633-1643
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  • 2025
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  • 2765-0189(pISSN)
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  • 2765-0235(eISSN)
아세아태평양축산학회 (Asian Australasian Association of Animal Production Societies)
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Serotonin promotes calcium accumulation and inhibits lipid accumulation in cultured goat mammary epithelial cells through HTR2A

  • ZhiFei Zhang (Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University) ;
  • BinHan Li (Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University) ;
  • YingFei Wang (Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University) ;
  • Tumaresi Abuduwufuer (Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University) ;
  • Kang Hu (Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University) ;
  • HuiLing Zheng (Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University)
  • 투고 : 2024.11.13
  • 심사 : 2025.03.13
  • 발행 : 2025.08.01
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초록

Objective: Fatty acids and calcium are both important nutrients in goat milk. Investigating the upstream molecular regulatory mechanisms that control the synthesis of milk fat and milk calcium in the mammary gland can help improve the quality of milk at its source. The objective of this study was to investigate the effects and regulatory pathways of serotonin (5-hydroxytryptamine, 5-HT) and its receptors on lipid synthesis and calcium ion levels in goat mammary epithelial cells (GMECs). Methods: GMECs isolated from live goats were treated with serotonin, overexpression of Serotonin receptor 2A (HTR2A), Sarpogrelate ([SAR] the specific antagonist of HTR2A), or a combination of these agents. The expression of genes related to de novo lipid synthesis in GMECs were detected using the Quantitative Real-Time polymerase chain reaction, the content of lipid droplets was detected using the BODIPY assay, and the calcium content was detected using the calcium chelating probe Fluo-3AM assay. Results: 5-HT dose-dependently promotes the activity of GEMCs, significantly inhibits the mRNA expression of key genes involved in de novo lipid synthesis such as ACC, FASN, SREBP1, SCD1 and ELOVL6 at a concentration of 100 μM, reduces triglyceride and total cholesterol content, suppresses lipid droplet accumulation in cells, and simultaneously promotes calcium accumulation in cells. Furthermore, overexpression of HTR2A in GMECs also induces an increase in cellular calcium levels and inhibits lipid synthesis and accumulation in cells. However, treatment of cells with SAR, the specific antagonist of HTR2A, significantly increases the levels of triglycerides, total cholesterol, and lipid droplet accumulation in cells. Conclusion: 5-HT inhibits lipid synthesis in GMECs while promoting an increase in cellular calcium levels, and this effect is mediated by the HTR2A receptor. Furthermorey, antagonists targeting HTR2A can reverse the inhibition of lipid synthesis and accumulation in cells.

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과제정보

This work was supported by the Key Projects of Shaanxi Natural Science Foundation Research Plan (grant no.: 2022JZ-10) and the Funding by Science and Technology Projects in Guangzhou (2024A04J0326).

참고문헌

  1. Angeles-Agdeppa I, Capanzana MV, Li-Yu J, Schollum LM, Kruger MC. High-calcium milk prevents overweight and obesity among postmenopausa women. Food Nutr Bull 2010;31:381-90. https://doi.org/10.1177/156482651003100301
  2. Gomes JMG, de Assis Costa J, Ribeiro PVM, Alfenas RCG. High calcium intake from fat-free milk, body composition and glycaemic control in adults with type 2 diabetes: a randomised crossover clinical trial. Br J Nutr 2019;122:301-8. https://doi.org/10.1017/S0007114519001259
  3. Suburu J, Shi L, Wu J, et al. Fatty acid synthase is required for mammary gland development and milk production during lactation. Am J Physiol Endocrinol Metab 2014;306:E1132-43. https://doi.org/10.1152/ajpendo.00514.2013
  4. Mamillapalli R, VanHouten J, Dann P, et al. Mammary specific ablation of the calcium-sensing receptor during lactation alters maternal calcium metabolism, milk calcium transport, and neonatal calcium accrual. Endocrinology 2013;154:3031-42. https://doi.org/10.1210/en.2012-2195
  5. Rapport MM, Green AA, Page IH. Serum vasoconstrictor (serotonin): III. chemical inactivation. J Biol Chem 1948;176:1237-41. https://doi.org/10.1016/S0021-9258(18)57136-2
  6. Berger M, Gray JA, Roth BL. The expanded biology of serotonin. Annu Rev Med 2009;60:355-66. https://doi.org/10.1146/annurev.med.60.042307.110802
  7. Sahu A, Gopalakrishnan L, Gaur N, et al. The 5-hydroxytryptamine signaling map: an overview of serotonin-serotonin receptor mediated signaling network. J Cell Commun Signal 2018;12:731-5. https://doi.org/10.1007/s12079-018-0482-2
  8. Matsuda M, Imaoka T, Vomachka AJ, et al. Serotonin regulates mammary gland development via an autocrine-paracrine loop. Dev Cell 2004;6:193-203. https://doi.org/10.1016/s1534-5807(04)00022-x
  9. Laporta J, Peters TL, Weaver SR, Merriman KE, Hernandez LL. Feeding 5-hydroxy-l-tryptophan during the transition from pregnancy to lactation increases calcium mobilization from bone in rats. Domest Anim Endocrinol 2013;44:176-84. https://doi.org/10.1016/j.domaniend.2013.01.005
  10. Connelly MK, Cheng AA, Hernandez LL. Graduate student literature review: serotonin and calcium metabolism: a story unfolding. J Dairy Sci 2021;104:13008-19. https://doi.org/10.3168/jds.2021-20610
  11. Zang WJ, Li H, Zhang ZF, et al. Serotonin induces parathyroid hormone-related protein in goat mammary gland. J Anim Sci 2018;96:1010-6. https://doi.org/10.1093/jas/skx023
  12. Zhang Z, Tian H, Chen X, et al. CRISPR/Cas9-mediated tryptophan hydroxylase 1 knockout decreases calcium transportation in goat mammary epithelial cells. Biochem Eng J 2021;171:108015. https://doi.org/10.1016/j.bej.2021.
  13. Laporta J, Moore SAE, Weaver SR, et al. Increasing serotonin concentrations alter calcium and energy metabolism in dairy COWS. J Endocrinol 2015;226:43-55. https://doi.org/10.1530/JOE-14-0693
  14. Choi W, Namkung], Hwang I, et al. Serotonin signals through a gut-liver axis to regulate hepatic steatosis. Nat Commun 2018;9:4824. https://doi.org/101038/s41467-018-07287-7 101038/s41467-018-07287-7
  15. Yun J, Jin H, Cao Y, et al. RNA-Seq analysis reveals a positive role of HTR2A in adipogenesis in Yan yellow cattle. Int J Mol Sci 2018;19:1760. https://doi.org/10.3390/ijms19061760
  16. Shi H, Shi H, Luo J, et al. Establishment and characterization of a dairy goat mammary epithelial cell line with human telomerase (hT-MECs). Anim Sci J 2014;85:735-43. https://doi.org/10.1111/asj.12206
  17. Chen X, Zhang Z, Niu H, et al. Goat milk improves glucose metabolism in type 2 diabetic mice and protects pancreatic B-cell functions. Mol Nutr Food Res 2024;68:2200842. https://doi.org/10.1002/mnfr.202200842
  18. Tian H, Luo J, Zhang Z, et al. CRISPR/Cas9-mediated stearoyl-CoA desaturase 1 (SCD1) deficiency affects fatty acid metabolism in goat mammary epithelial cells. J Agric Food Chem 2018;66:10041-52. https://doi.org/10.1021/acs.jafc.8b03545
  19. Namkung J, Shong KE, Kim H, Oh CM, Park S, Kim H. Inhibition of serotonin synthesis induces negative hepatic lipid balance. Diabetes Metab 11 2018;42:233-43. https://doi.org/10.4093/dmj.2017.0084
  20. Crane JD, Palanivel R, Mottillo EP, et al. Inhibiting peripheral serotonin synthesis reduces obesity and metabolic dysfunction by promoting brown adipose tissue thermogenesis. Nat Med 2015;21:166-72. https://doi.org/10.1038/nm.3766
  21. Horseman ND, Collier RJ. Serotonin: a local regulator in the mammary gland epithelium. Annu Rev Anim Biosci 2014;2:353-74. https://doi.org/10.1146/annurev-animal-022513-114227
  22. Wang L, Fan X, Han J, et al. Gut-derived serotonin contributes to the progression of non-alcoholic steatohepatitis via the liver HTR2A/PPARgamma2 pathway. Front Pharmacol 2020;11:553. https://doi.org/10.3389/fphar.2020.00553
  23. Sumara G, Sumara O, Kim JK, Karsenty G. Gut-derived sero-tonin is a multifunctional determinant to fasting adaptation. Cell Metab 2012;16:588-600. https://doi.org/10.1016/j.cmet.2012.09.014
  24. Hansen HO, Tornehave D, Knudsen J. Synthesis of milkspecific fatty acids and proteins by dispersed goat mammary-gland epithelial cells. Biochem J 1986;238:167-72. https://doi.org/10.1042/bj2380167
  25. Li P, Fang X, Hao G, et al. Methionine promotes milk protein synthesis via the PI3K-mTOR signaling pathway in human mammary epithelial cells. Metabolites 2023;13:1149. https://doi.org/10.3390/metabol3111149
  26. Akersr MR, Bauman DE, Capuco AV, Goodman GT, Tucker AH. Prolactin regulation of milk secretion and biochemical differentiation of mammary epithelial cells in periparturient COWS. Endocrinology 1981;109:23-30. https://doi.org/10.1210/endo-109-1-23
  27. Cameron CM, Rillema JA. Extracellular calcium ion concen-tration required for prolactin to express its actions on casein, ribonucleic acid, and lipid biosynthesis in mouse mammary gland explants. Endocrinology 1983;113:1596-600. https://doi.org/10.1210/endo-113-5-1596
  28. Zhao H, Chen S, Hu K, et al. 5-HTP decreases goat mammary epithelial cells apoptosis through MAPK/ERK/Bd-pathway. Gene 2021;769145240. https://doi.org/10.1016/j.gene.2020.145240
  29. Chen S, Zhao H, Yan X, et al. 5-Hydroxy-1-tryptophan promotes the milk calcium level via the miR-99a-3p/ATP2B1 axis in goat mammary epithelial cells. 3 Agric Food Chem 2020;68:3277-85. https://doi.org/10.1021/acs.jafc.9b07869
  30. Suarez-Truillo A, Argiiello A, Rivero MA, Capote], Castro N. Short communication: differences in distribution of serotonin receptor subtypes in the mammary gland of sheep, goats, and COWS during lactation and involution. J Dairy Sci 2019;102:2703-7. https://doi.org/10.3168/jds.2018-15328
  31. Pai VP, Hernandez LL, Stull MA, Horseman ND. The type 7 serotonin receptor, 5-HT7, is essential in the mammary gland for regulation of mammary epithelial structure and function. BioMed Res Int 2015;2015:364746. https://doi.org/10.1155/2015/364746
  32. Fawley JA, Doyle MW, Andresen MC. 5-HT3R-sourced calcium enhances glutamate release from a distinct vesicle pool. Brain Res 2019;1721:146346. https://doi.org/10.1016/j.brainres.2019.146346
  33. Turner JH, Raymond JR. Interaction of calmodulin with the serotonin 5-hydroxytryptamine 2A receptor: a putative regulator of G protein coupling and receptor phosphorylation by protein kinase C. J Biol Chem 2005;280:30741-50. https://doi.org/101074/jbc.M501696200 101074/jbc.M501696200
  34. Farrelly LA, Thompson RE, Zhao S, et al. Histone serotonylation is a permissive modification that enhances TFIID binding to H3K4me3. Nature 2019;567:535-9. https://doi.org/10.1038/s41586-019-1024-7
  35. Al-Kachak A, Fulton SL. DiSalvo G, et al. Histone H3 serotonylation dynamics in dorsal raphe nucleus contribute to stress- and antidepressant-mediated gene expression and behavior. bioRxiv 539464 [Preprint]. 2023 [cited 2025 Jan 12]. https://doi.org/10.1101/2023.05.04.539464