Proteomics and Structural Biology Lab, Animal Biotechnology Center, National Dairy Research Institute, Karnal, 132001, Haryana, India.
Department of Veterinary Physiology and Biochemistry, Veterinary College and Research Institute (TANUVAS), Orathanadu, 614625, Tamilnadu, India.
Sci Rep. 2020 Mar 16;10(1):4834. doi: 10.1038/s41598-020-61521-1.
The mature mammary gland is made up of a network of ducts that terminates in alveoli. The innermost layer of alveoli is surrounded by the differentiated mammary epithelial cells (MECs), which are responsible for milk synthesis and secretion during lactation. However, the MECs are in a state of active proliferation during pregnancy, when they give rise to network like structures in the mammary gland. Buffalo (Bubalus bubalis) constitute a major source of milk for human consumption, and the MECs are the major precursor cells which are mainly responsible for their lactation potential. The proteome of MECs defines their functional state and suggests their role in various cellular activities such as proliferation and lactation. To date, the proteome profile of MECs from buffalo origin is not available. In the present study, we have profiled in-depth proteome of in vitro cultured buffalo MECs (BuMECs) during active proliferation using high throughput tandem mass spectrometry (MS). MS analysis identified a total of 8330, 5970, 5289, 4818 proteins in four sub-cellular fractions (SCFs) that included cytosolic (SCF-I), membranous and membranous organelle's (SCF-II), nuclear (SCF-III), and cytoskeletal (SCF-IV). However, 792 proteins were identified in the conditioned media, which represented the secretome. Altogether, combined analysis of all the five fractions (SCFs- I to IV, and secretome) revealed a total of 12,609 non-redundant proteins. The KEGG analysis suggested that these proteins were associated with 325 molecular pathways. Some of the highly enriched molecular pathways observed were metabolic, MAPK, PI3-AKT, insulin, estrogen, and cGMP-PKG signalling pathway. The newly identified proteins in this study are reported to be involved in NOTCH signalling, transport and secretion processes.
成熟的乳腺由导管网络组成，其末端为腺泡。腺泡的最内层被分化的乳腺上皮细胞（MEC）包围，这些细胞在哺乳期负责乳汁的合成和分泌。然而，在怀孕期间，MEC 处于活跃增殖状态，此时它们在乳腺中产生网状结构。水牛（Bubalus bubalis）是人类消费牛奶的主要来源，MEC 是主要的前体细胞，主要负责其泌乳潜力。MEC 的蛋白质组定义了它们的功能状态，并表明它们在各种细胞活动中的作用，如增殖和泌乳。迄今为止，水牛来源的 MEC 的蛋白质组图谱尚不可用。在本研究中，我们使用高通量串联质谱（MS）对体外培养的水牛 MEC（BuMEC）在活跃增殖期间进行了深入的蛋白质组分析。MS 分析共鉴定了 8330、5970、5289、4818 种蛋白质，这些蛋白质分别位于四个亚细胞区室（SCF）中，包括细胞质（SCF-I）、膜和膜细胞器（SCF-II）、核（SCF-III）和细胞骨架（SCF-IV）。然而，在条件培养基中鉴定出了 792 种蛋白质，代表了分泌组。总的来说，对所有五个区室（SCF-I 到 IV 和分泌组）的联合分析共揭示了 12609 种非冗余蛋白质。KEGG 分析表明，这些蛋白质与 325 个分子途径相关。观察到的一些高度富集的分子途径是代谢、MAPK、PI3-AKT、胰岛素、雌激素和 cGMP-PKG 信号通路。本研究中新鉴定的蛋白质被报道参与 NOTCH 信号转导、运输和分泌过程。
