File(s) under permanent embargo
Copper and lactational hormones influence the CTR1 copper transporter in PMC42-LA mammary epithelial cell culture models
journal contribution
posted on 2014-04-01, 00:00 authored by David Freestone, Michael Cater, Leigh AcklandLeigh Ackland, D Paterson, D L Howard, M D de Jonge, Agnes MichalczykAgnes MichalczykAdequate amounts of copper in milk are critical for normal neonatal development, however the mechanisms regulating copper supply to milk have not been clearly defined. PMC42-LA cell cultures representative of resting, lactating and suckled mammary epithelia were used to investigate the regulation of the copper uptake protein, CTR1. Both the degree of mammary epithelial differentiation (functionality) and extracellular copper concentration greatly impacted upon CTR1 expression and its plasma membrane association. In all three models (resting, lactating and suckling) there was an inverse correlation between extracellular copper concentration and the level of CTR1. Cell surface biotinylation studies demonstrated that as extracellular copper concentration increased membrane associated CTR1 was reduced. There was a significant increase in CTR1 expression (total and membrane associated) in the suckled gland model in comparison to the resting gland model, across all copper concentrations investigated (0-50 μM). Regulation of CTR1 expression was entirely post-translational, as quantitative real-time PCR analyses showed no change to CTR1 mRNA between all models and culture conditions. X-ray fluorescence microscopy on the differentiated PMC42-LA models revealed that organoid structures distinctively accumulated copper. Furthermore, as PMC42-LA cell cultures became progressively more specialised, successively more copper accumulated in organoids (resting
History
Journal
Journal of Nutritional BiochemistryVolume
25Issue
4Pagination
377 - 387Publisher
ElsevierLocation
New York, NYPublisher DOI
eISSN
1873-4847Language
engPublication classification
C Journal article; C1 Refereed article in a scholarly journalCopyright notice
2014, ElsevierUsage metrics
Categories
No categories selectedKeywords
CTR1CopperHuman mammary glandLactationMetallothioneinX-ray fluorescence microscopyScience & TechnologyLife Sciences & BiomedicineBiochemistry & Molecular BiologyNutrition & DieteticsMETHIONINE-RICH CLUSTERSINTESTINAL CACO-2 CELLSHUMAN-BREASTSTIMULATED ENDOCYTOSISMAMMALIAN-CELLSCARCINOMA CELLSTOXIC MILKLINE PMC42HCTR1EXPRESSIONGLANDLOCALIZATIONCISPLATINATP7B
Licence
Exports
RefWorks
BibTeX
Ref. manager
Endnote
DataCite
NLM
DC