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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: 877931004@qq.com
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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 

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