Hostname: page-component-cd9895bd7-fscjk Total loading time: 0 Render date: 2024-12-26T12:20:12.217Z Has data issue: false hasContentIssue false
  • English
  • Français

Milk fat globule is an alternative to mammary epithelial cells for gene expression analysis in buffalo

Published online by Cambridge University Press:  01 April 2016

Qiuming Chen ,
Yanjun Wu ,
Mingyuan Zhang ,
Wenwen Xu ,
Xiaoping Guo ,
Xueyu Yan ,
Haiying Deng ,
Qinyang Jiang ,
Xiurong Yang  and
Ganqiu Lan
...Show all authors
Qiuming Chen*
Affiliation:
College of Animal Science and Technology, Guangxi University, Nanning 530004, China
Yanjun Wu
Affiliation:
College of Animal Science and Technology, Guangxi University, Nanning 530004, China
Mingyuan Zhang
Affiliation:
College of Animal Science and Technology, Guangxi University, Nanning 530004, China
Wenwen Xu
Affiliation:
College of Animal Science and Technology, Guangxi University, Nanning 530004, China
Xiaoping Guo
Affiliation:
College of Animal Science and Technology, Guangxi University, Nanning 530004, China
Xueyu Yan
Affiliation:
College of Animal Science and Technology, Guangxi University, Nanning 530004, China
Haiying Deng
Affiliation:
College of Animal Science and Technology, Guangxi University, Nanning 530004, China
Qinyang Jiang
Affiliation:
College of Animal Science and Technology, Guangxi University, Nanning 530004, China
Xiurong Yang
Affiliation:
College of Animal Science and Technology, Guangxi University, Nanning 530004, China
Ganqiu Lan
Affiliation:
College of Animal Science and Technology, Guangxi University, Nanning 530004, China
Yafen Guo
Affiliation:
College of Animal Science and Technology, Guangxi University, Nanning 530004, China
Guangsheng Qin
Affiliation:
Guangxi Key Laboratory of Buffalo Genetics and Breeding, Guangxi Buffalo Research Institute, Chinese Academy of Agriculture Science, Nanning 530004, China
Hesheng Jiang
Affiliation:
College of Animal Science and Technology, Guangxi University, Nanning 530004, China
*
*For correspondence; e-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

Owing to the difficulty in obtaining mammary gland tissue from lactating animals, it is difficult to test the expression levels of genes in mammary gland. The aim of the current study was to identify if milk fat globule (MFG) in buffalo milk was an alternative to mammary gland (MG) and milk somatic cell (MSC) for gene expression analysis. Six buffalos in late lactation were selected to collect MFG and MSC, and then MG was obtained by surgery. MFG was stained with acridine orange to successfully visualise RNA and several cytoplasmic crescents in MFG. The total RNA in MFG was successfully isolated and the integrity was assessed by agarose gel electrophoresis. We analysed the cellular components in MFG, MG and MSC through testing the expression of cell-specific genes by qRT-PCR. The results showed that adipocyte-specific gene (AdipoQ) and leucocyte-specific genes (CD43, CSF1 and IL1α) in MFG were not detected, whereas epithelial cell marker genes (Keratin 8 and Keratin 18) in MFG were higher than in MSC and lower than in MG, fibroblast marker gene (vimentin) in MFG was significantly lower than in MG and MSC, milk protein genes (LALBA, BLG and CSN2) and milk fat synthesis-related genes (ACC, BTN1A1, FABP3 and FAS) in MFG were higher than in MG and MSC. In conclusion, the total RNA in MFG mainly derives from mammary epithelial cells and can be used to study the functional gene expression of mammary epithelial cells.

Keywords

Buffalomilk fat globulemammary epithelial cellmilk somatic cell
Type
Research Article
Information
Journal of Dairy Research , Volume 83 , Issue 2 , May 2016 , pp. 202 - 208
Copyright
Copyright © Proprietors of Journal of Dairy Research 2016 

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

Avondo, M, Pennisi, P, Lanza, M, Pagano, RI, Valenti, B, Di Gregorio, P, De Angelis, A, Giorgio, D & Di Trana, A 2015 Effect of the αs 1-casein genotype and its interaction with diet degradability on milk production, milk quality, metabolic and endocrinal response of Girgentana goats. Small Ruminant Research 123 136–141Google Scholar
Bauman, DE & Griinari, JM 2003 Nutritional regulation of milk fat synthesis. Annual Review of Nutrition 23 203–227Google Scholar
Bionaz, M & Loor, JJ 2007 Identification of reference genes for quantitative real-time PCR in the bovine mammary gland during the lactation cycle. Physiological Genomics 29 312–319Google Scholar
Bionaz, M & Loor, JJ 2008 Gene networks driving bovine milk fat synthesis during the lactation cycle. BMC Genomics 9 366CrossRefGoogle ScholarPubMed
Boutinaud, M & Jammes, H 2002 Potential uses of milk epithelial cells: a review. Reproduction Nutrition Development 42 133–147Google Scholar
Boutinaud, M, Rulquin, H, Keisler, D, Djiane, J & Jammes, H 2002 Use of somatic cells from goat milk for dynamic studies of gene expression in the mammary gland. Journal of Animal Science 80 1258–1269CrossRefGoogle ScholarPubMed
Brenaut, P, Bangera, R, Bevilacqua, C, Rebours, E, Cebo, C & Martin, P 2012 Validation of RNA isolated from milk fat globules to profile mammary epithelial cell expression during lactation and transcriptional response to a bacterial infection. Journal of Dairy Science 95 6130–6144Google Scholar
Bustin, SA, Benes, V, Garson, JA, Hellemans, J, Huggett, J, Kubista, M, Mueller, R, Nolan, T, Pfaffl, MW & Shipley, GL 2009 The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clinical Chemistry 55 611–622CrossRefGoogle ScholarPubMed
Cánovas, A, Rincรณn, G, Bevilacqua, C, Islas-Trejo, A, Brenaut, P, Hovey, RC, Boutinaud, M, Morgenthaler, C, VanKlompenberg, MK & Martin, P 2014 Comparison of five different RNA sources to examine the lactating bovine mammary gland transcriptome using RNA-Sequencing. Scientific Reports 4 5297Google Scholar
Capuco, A, Wood, D, Baldwin, R, McLeod, K & Paape, M 2001 Mammary cell number, proliferation, and apoptosis during a bovine lactation: relation to milk production and effect of bST. Journal of Dairy Science 84 2177–2187CrossRefGoogle ScholarPubMed
Cerón-Muñoz, M, Tonhati, H, Duarte, J, Oliveira, J, Muñoz-Berrocal, M & Jurado-Gámez, H 2002 Factors affecting somatic cell counts and their relations with milk and milk constituent yield in buffaloes. Journal of Dairy Science 85 2885–2889Google Scholar
Choudhary, RK, Kaur, H, Choudhary, S & Verma, R 2015 Distribution and analysis of milk fat globule and crescent in murrah buffalo and crossbred cow. Proceedings of the National Academy of Sciences, India Section B: Biological Sciences 85 1–6Google Scholar
Cui, X, Hou, Y, Yang, S, Xie, Y, Zhang, S, Zhang, Y, Zhang, Q, Lu, X, Liu, GE & Sun, D 2014 Transcriptional profiling of mammary gland in Holstein cows with extremely different milk protein and fat percentage using RNA sequencing. BMC Genomics 15 226CrossRefGoogle ScholarPubMed
Hu, H, Wang, J, Bu, D, Wei, H, Zhou, L, Li, F & Loor, JJ 2009 In vitro culture and characterization of a mammary epithelial cell line from Chinese Holstein dairy cow. PLoS ONE 4(11) e7636CrossRefGoogle ScholarPubMed
Huston, GE & Patton, S 1990 Factors related to the formation of cytoplasmic crescents on milk fat globules. Journal of Dairy Science 73 2061–2066Google Scholar
Jeong, J, Rao, AU, Xu, J, Ogg, SL, Hathout, Y, Fenselau, C & Mather, IH 2009 The PRY/SPRY/B30. 2 Domain of Butyrophilin 1A1 (BTN1A1) Binds to Xanthine Oxidoreductase implications for the function of BTN1A1 in the mammary gland and other tissues. Journal of Biological Chemistry 284 22444–22456CrossRefGoogle ScholarPubMed
Keenan, T & Mather, I 2006 Intracellular origin of milk fat globules and the nature of the milk fat globule membrane. In Fox PF & McSweeney PLH, Editors, Advanced Dairy Chemistry Volume 2 Lipids, pp. 137–171. New York: SpringerCrossRefGoogle Scholar
Liang, M-y, Hou, X-m, Qu, B, Zhang, N, Li, N, Cui, Y-j, Li, Q-z & Gao, X-J 2014 Functional analysis of FABP3 in the milk fat synthesis signaling pathway of dairy cow mammary epithelial cells. In Vitro Cellular and Developmental Biology-Animal 50 865–873CrossRefGoogle ScholarPubMed
Lindmark-Mansson, H, Branning, C, Alden, G & Paulsson, M 2006 Relationship between somatic cell count, individual leukocyte populations and milk components in bovine udder quarter milk. International Dairy Journal 16 717–727CrossRefGoogle Scholar
Lu, J, van Hooijdonk, T, Boeren, S, Vervoort, J & Hettinga, K 2014 Identification of lipid synthesis and secretion proteins in bovine milk. Journal of Dairy Research 81 65–72Google Scholar
Maningat, PD, Sen, P, Sunehag, AL, Hadsell, DL & Haymond, MW 2007 Regulation of gene expression in human mammary epithelium: effect of breast pumping. Journal of Endocrinology 195 503–511Google Scholar
Maningat, PD, Sen, P, Rijnkels, M, Sunehag, AL, Hadsell, DL, Bray, M & Haymond, MW 2009 Gene expression in the human mammary epithelium during lactation: the milk fat globule transcriptome. Physiological Genomics 37 12–22CrossRefGoogle ScholarPubMed
Maningat, PD, Sen, P, Rijnkels, M, Hadsell, DL, Bray, MS & Haymond, MW 2011 Short-term administration of rhGH increases markers of cellular proliferation but not milk protein gene expression in normal lactating women. Physiological Genomics 43 381–391Google Scholar
Mohammad, MA & Haymond, MW 2013 Regulation of lipid synthesis genes and milk fat production in human mammary epithelial cells during secretory activation. American Journal of Physiology 305 E700–E716Google Scholar
Mohammad, MA, Hadsell, DL & Haymond, MW 2012 Gene regulation of UDP-galactose synthesis and transport: potential rate-limiting processes in initiation of milk production in humans. American Journal of Physiology 303 E365–E376Google Scholar
Murrieta, C, Hess, B, Scholljegerdes, E, Engle, T, Hossner, K, Moss, G & Rule, D 2006 Evaluation of milk somatic cells as a source of mRNA for study of lipogenesis in the mammary gland of lactating beef cows supplemented with dietary high-linoleate safflower seeds. Journal of Animal Science 84 2399–2405Google Scholar
Rudolph, MC, McManaman, JL, Hunter, L, Phang, T & Neville, MC 2003 Functional development of the mammary gland: use of expression profiling and trajectory clustering to reveal changes in gene expression during pregnancy, lactation, and involution. Journal of Mammary Gland Biology and Neoplasia 8 287–307Google Scholar
Rudolph, MC, Neville, MC & Anderson, SM 2007 Lipid synthesis in lactation: diet and the fatty acid switch. Journal of Mammary Gland Biology and Neoplasia 12 269–281Google Scholar
Stewart, M 1993 Intermediate filament structure and assembly. Current Opinion in Cell Biology 5 3–11Google Scholar
Thomas, CS, Svennersten-Sjaunja, K, Bhosrekar, MR & Bruckmaier, RM 2004 Mammary cisternal size, cisternal milk and milk ejection in Murrah buffaloes. Journal of Dairy Research 71 162–168Google Scholar
Thompson, P, Kadlubar, F, Vena, S, Hill, H, McClure, G, McDaniel, L & Ambrosone, C 1998 Exfoliated ductal epithelial cells in human breast milk: a source of target tissue DNA for molecular epidemiologic studies of breast cancer. Cancer Epidemiology Biomarkers and Prevention 7 37–42Google Scholar
Wickramasinghe, S, Rincon, G, Islas-Trejo, A & Medrano, JF 2012 Transcriptional profiling of bovine milk using RNA sequencing. BMC Genomics 13 45CrossRefGoogle ScholarPubMed
Xavier, M, Paixรฃo, T, Poester, F, Lage, A & Santos, R 2009 Pathological, immunohistochemical and bacteriological study of tissues and milk of cows and fetuses experimentally infected with Brucella abortus. Journal of Comparative Pathology 140 149–157Google Scholar
Yadav, P, Singh, DD, Mukesh, M, Kataria, R, Yadav, A, Mohanty, A & Mishra, B 2012 Identification of suitable housekeeping genes for expression analysis in mammary epithelial cells of buffalo (Bubalus bubalis) during lactation cycle. Livestock Science 147 72–76Google Scholar
Zhao, X, He, W, Song, Z, Tong, Z, Li, S & Ni, L 2012 Mineral trioxide aggregate promotes odontoblastic differentiation via mitogen-activated protein kinase pathway in human dental pulp stem cells. Molecular Biology Reports 39 215–220Google Scholar