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Lookup NU author(s): Iain Haslam, Professor Nicholas Simmons
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Rumen fermentation of plant-based forage in bovines is the major site for generation and absorption of short-chain fatty acids. Consequentially, the rumen is also the site for initial exposure to toxins released from diet. Accordingly, we have investigated the expression of bovine ABC transporters in the rumen associated with cytoprotection against xenobiotic exposure, namely MDR1 (ABCB1), MRP2 (ABCC2) and BCRP (ABCG2). Bovine rumen samples from the ventral sac were obtained post-mortem from a commercial slaughterhouse after humane killing. Rumen papilla samples were then prepared for total RNA isolation for RT-PCR, SDS-PAGE/Western blotting and immunohistochemistry. PCR products of the predicted size were observed for both MDR1 and BCRP, but not for MRP2 using bovine-specific primers. beta-actin was used as a control transcript. Western blot analysis using C219 primary monoclonal antibody revealed MDR1 protein expression in bovine rumen (Mapp, of similar to 170-180 kD). Immunolocalisation of MDR1 using UIC2 monoclonal antibody within cryosections of bovine rumen showed extensive membrane staining in the cells of the stratum granulosum, stratum spinosum and stratum basale. MDR1 expression was absent from outer stratum corneum. Protein expression and immunolocalisation were also confirmed for BCRP, with prevalent staining in the stratum basale, becoming weaker in the stratum spinosum and stratum granulosum.
Author(s): Haslam IS, Simmons NL
Publication type: Article
Publication status: Published
Journal: Journal of Comparative Physiology B
Year: 2014
Volume: 184
Issue: 5
Pages: 673-681
Print publication date: 01/07/2014
Online publication date: 20/04/2014
Acceptance date: 13/01/2014
ISSN (print): 0174-1578
ISSN (electronic): 1432-136X
Publisher: Springer
URL: http://dx.doi.org/10.1007/s00360-014-0804-5
DOI: 10.1007/s00360-014-0804-5
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