Abstract
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.
Highlights
The mature mammary gland is made up of a network of ducts that terminates in alveoli
sub-cellular fractions (SCFs) which included cytosolic (SCF-I); membranous and membranous organelles (SCF-II); nuclear (SCF-III); and cytoskeletal (SCF-IV)], and conditioned media, from actively proliferating buffalo MECs (BuMECs) were analyzed for identification of proteins using the mass spectrometry (MS)
The MS analysis of conditioned media identified a total of 792 non-redundant proteins (Table S5), which represent the secretome of actively proliferating BuMECs
Summary
The mature mammary gland is made up of a network of ducts that terminates in alveoli. We performed in-depth proteome profiling of actively proliferating BuMECs. Ruminants (cattle, buffalo, goat, etc.) constitute a distinct class of livestock species, which are known milk producers. Given the differences in mammary gland physiology between ruminants and non-ruminants, expression of various proteins and molecular pathways associated with proliferation and lactogenesis, mimicking in vivo mammary gland development and lactation respectively, may be different in BuMECs. To our knowledge, no information is available on the proteome of BuMECs. we have performed proteome analysis of 68 digested protein fractions [24 digested protein fractions for sub-cellular fraction-I (SCF-I); 12 digested protein fractions- each for SCF-II, -III, and -IV; and 8 digested protein fractions for conditioned media] to generate in-depth proteome data, which is the uniqueness of this study. The current dataset on BuMECs proteome will add to the existing information available on mammary proteome and, constitute a reservoir of proteins for further investigation and characterization of the ruminant mammary system in general, the buffalo
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
