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Use of milk epithelial cells to study regulation of cell activity and apoptosis during once-daily milking in goats

Published online by Cambridge University Press:  09 November 2010

H. Ben Chedly
Affiliation:
INRA, UMR1080 Production du Lait, F-35590 Saint-Gilles, France Agrocampus Ouest, UMR1080 Production du Lait, F-35000 Rennes, France
P. Lacasse
Affiliation:
Dairy and Swine R&D Centre, Agriculture and Agri-Food Canada, PO Box 90 STN Lennoxville, Sherbrooke, Quebec, Canada J1M 1Z3
P.-G. Marnet
Affiliation:
INRA, UMR1080 Production du Lait, F-35590 Saint-Gilles, France Agrocampus Ouest, UMR1080 Production du Lait, F-35000 Rennes, France
M. Komara
Affiliation:
INRA, UMR1080 Production du Lait, F-35590 Saint-Gilles, France Agrocampus Ouest, UMR1080 Production du Lait, F-35000 Rennes, France
S. Marion
Affiliation:
INRA, UMR1080 Production du Lait, F-35590 Saint-Gilles, France Agrocampus Ouest, UMR1080 Production du Lait, F-35000 Rennes, France
M. Boutinaud*
Affiliation:
INRA, UMR1080 Production du Lait, F-35590 Saint-Gilles, France Agrocampus Ouest, UMR1080 Production du Lait, F-35000 Rennes, France
*
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Abstract

Generally, once-daily milking (ODM) decreases milk yield. This effect may be the consequence of a decrease in mammary epithelial cell (MEC) activity or a reduction in their number. The aim of this study was to determine the effect of ODM on the synthetic activity and rate of apoptosis of MEC using a non-invasive method. Eight Alpine goats were subjected to ODM or twice-daily milking for two 5-week periods. MECs were purified by centrifugation and immunocytochemical binding in milk after 1 and 5 weeks of each period. mRNA levels of some proteins involved in lactose and milk protein synthesis and in apoptosis were evaluated using real-time PCR. Isolation of MEC from milk was a useful method to investigate transcriptional regulation in a timeline study. ODM induced greater decreases in milk, lactose and protein yields after 1 week than after 5 weeks. This suggests an adaptation of the mammary gland to ODM, which reduces the inhibitory effect of this practice. Reductions in milk component yields were associated with lower α-lactalbumin transcripts, suggesting a transcriptional decrease of lactose synthesis during ODM. Glucose transporter GLUT1 transcripts were downregulated under ODM, suggesting that lactose precursor uptake by MEC might be involved in the regulation of lactose synthesis. κ-Casein mRNA levels tended to be lower during ODM. ODM increased levels of the pro-apoptotic transcript Bax after both 1 and 5 weeks, but no variation was observed in the Bax/Bcl-2 ratio. ODM affected cell synthetic activity through transcriptional regulation and may have induced apoptosis. The reduction of the negative effect of ODM on milk yield suggests that Alpine goats are able to adapt to ODM. Further studies are needed to investigate the effect of ODM on MEC turnover.

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Full Paper
Copyright
Copyright © The Animal Consortium 2010

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References

Andersen, CL, Jensen, JL, Orntoft, TF 2004. Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Research 64, 52455250.CrossRefGoogle Scholar
Ben Chedly, H, Lacasse, P, Marnet, PG, Wiart-Letort, S, Finot, L, Boutinaud, M 2009. Cell junction disruption after 36 h milk accumulation was associated with changes to mammary secretory tissue activity and dynamics in lacting dairy goats. Journal of Physiology and Pharmacology 60 (suppl. 3), 105111.Google Scholar
Bernier-Dodier, P, Delbecchi, L, Wagner, GF, Talbot, BG, and Lacasse, P 2010. Effect of milking frequency on lactation persistency and mammary gland remodelling in mid-lactation cows. Journal of Dairy Science 93, 555564.CrossRefGoogle ScholarPubMed
Boutinaud, M, Jammes, H 2002. Potential uses of milk epithelial cells: a review. Reproduction Nutrition Development 42, 133147.CrossRefGoogle ScholarPubMed
Boutinaud, M, Galio, L, Devinoy, E 2009. Transcripts in milk purified mammary epithelial cells can reveal the effect of once-daily milking. 6th International Symposium of Milk Genomics and Human Health, Paris, France.Google Scholar
Boutinaud, M, Rousseau, C, Keisler, DH, Jammes, H 2003. Growth hormone and milking frequency act differently on goat mammary gland in late lactation. Journal of Dairy Science 86, 509520.CrossRefGoogle ScholarPubMed
Boutinaud, M, Ben Chedly, MH, Delamaire, E, Guinard-Flament, J 2008. Milking and feed restriction regulate transcripts of mammary epithelial cells purified from milk. Journal of Dairy Science 91, 988998.CrossRefGoogle ScholarPubMed
Boutinaud, M, Rulquin, H, Keisler, DH, Djiane, J, Jammes, H 2002. Use of somatic cells from goat milk for dynamic studies of gene expression in the mammary gland. Journal of Animal Science 80, 12581269.CrossRefGoogle ScholarPubMed
Boutinaud, M, Shand, JH, Park, MA, Phillips, K, Beattie, J, Flint, DJ, Allan, GJ 2004. A quantitative RT-PCR study of the mRNA expression profile of the IGF axis during mammary gland development. Journal of Molecular Endocrinology 33, 195207.CrossRefGoogle ScholarPubMed
Bryson, JM, Wilde, CJ, Addey, CV 1993. Effect of unilateral changes in milking frequency on mammary mRNA concentrations in the lactating goat. Biochemical Society Transactions 21 (Part 3), 294S.CrossRefGoogle ScholarPubMed
Camps, M, Vilaro, S, Testar, X, Palacin, M, Zorzano, A 1994. High and polarized expression of Glut1 glucose transporters in epithelial-cells from mammary-gland – acute down-regulation of Glut1 carriers by weaning. Endocrinology 134, 924934.CrossRefGoogle ScholarPubMed
Capuco, AV, Ellis, SE, Hale, SA, Long, E, Erdman, RA, Zhao, X, Paape, MJ 2003. Lactation persistency: insights from mammary cell proliferation studies. Journal of Animal Science 81, 1831.CrossRefGoogle ScholarPubMed
Castillo, V, Such, X, Caja, G, Casals, R, Albanell, E, Salama, AAK 2008. Effect of milking interval on milk secretion and mammary tight junction permeability in dairy ewes. Journal of Dairy Science 91, 26102619.CrossRefGoogle ScholarPubMed
Delamaire, E, Guinard-Flament, J 2006. Increasing milking intervals decreases the mammary blood flow and mammary uptake of nutrients in dairy cows. Journal of Dairy Science 89, 34393446.CrossRefGoogle ScholarPubMed
Farr, V, Stelwagen, K, Kerr, M, Davis, S, Eichler, S 1995. Effect of once daily milking (ODM) on enzyme activities in the bovine mammary gland. Proceedings of the New Zealand Society of Animal Production 55, 1213.Google Scholar
Feng, S, Salter, AM, Parr, T, Garnsworthy, PC 2007. Extraction and quantitative analysis of stearoyl-coenzyme A desaturase mRNA from dairy cow milk somatic cells. Journal of Dairy Science 90, 41284136.CrossRefGoogle ScholarPubMed
Hayashi, AA, McCoard, SA, Roy, NC, Barnett, MPG, Mackenzie, DDS, Mcnabb, WC 2004. Gene expression in bovine mammary somatic cells isolated from milk. Journal of Animal and Feed Sciences 13, 401404.CrossRefGoogle Scholar
INRA 2007. Ruminant nutrition: recommended allowances and feed tables. Quae, Versailles, France.Google Scholar
Kelly, AL, Reid, S, Joyce, P, Meaney, WJ, Foley, J 1998. Effect of decreased milking frequency of cows in late lactation on milk somatic cell count, polymorphonuclear leucocyte numbers, composition and proteolytic activity. Journal of Dairy Research 65, 365373.CrossRefGoogle ScholarPubMed
Knight, CH, Wilde, CJ 1993. Mammary cell changes during pregnancy and lactation. Livestock Production Science 35, 319.CrossRefGoogle Scholar
Komara, M, Boutinaud, M, Ben Chedly, H, Guinard-Flament, J, Marnet, PG 2009. Once daily milking effects in high yielding alpine dairy goats. Journal of Dairy Science 92, 54475455.CrossRefGoogle ScholarPubMed
Lacy-Hulbert, SJ, Woolford, MW, Nicholas, GD, Prosser, CG, Stelwagen, K 1999. Effect of milking frequency and pasture intake on milk yield and composition of late lactation cows. Journal of Dairy Science 82, 12321239.CrossRefGoogle ScholarPubMed
Li, P, Rudland, PS, Fernig, DG, Finch, LM, Wilde, CJ 1999. Modulation of mammary development and programmed cell death by the frequency of milk removal in lactating goats. Journal of Physiology 519 (Part 3), 885900.CrossRefGoogle ScholarPubMed
Marnet, PG, Komara, M 2008. Management systems with extended milking intervals in ruminants: regulation of production and quality of milk. Journal of Animal Science 86, 4756.CrossRefGoogle ScholarPubMed
Murrieta, CM, Hess, BW, Scholljegerdes, EJ, Engle, TE, Hossner, KL, Moss, GE, Rule, DC 2006. Evaluation of milk somatic cells as a source of mRNA for study of lipogenesis in the mammary gland of lactating beef cows supplemented with dietary high-linoleate safflower seeds. Journal of Animal Science 84, 23992405.CrossRefGoogle ScholarPubMed
Nudda, A, Bencini, R, Mijatovic, S, Pullina, G 2002. The yield and composition of milk in Sarda, Awassi, and Merino sheep milked unilaterally at different frequencies. Journal of Dairy Science 85, 28792884.CrossRefGoogle ScholarPubMed
Pfaffl, MW, Tichopad, A, Prgomet, C, Neuvians, TP 2004. Determination of stable housekeeping genes, differentially regulated target genes and sample integrity: BestKeeper–Excel-based tool using pair-wise correlations. Biotechnology Letters 26, 509515.CrossRefGoogle ScholarPubMed
Pomies, D, Remond, B, Pradel, P 2004. Performances and milk quality of dairy cows temporary milked once a day: incidence of once daily milking (ODM) duration and lactation stage. Rencontres Recherches Ruminants 11, 225228.Google Scholar
Salama, AA, Such, X, Caja, G, Rovai, M, Casals, R, Albanell, E, Marin, MP, Marti, A 2003. Effects of once versus twice daily milking throughout lactation on milk yield and milk composition in dairy goats. Journal of Dairy Science 86, 16731680.CrossRefGoogle ScholarPubMed
SAS Institute 1999. Statistical Analysis System Release 8.01. Cary, NC, USA.Google Scholar
Sorensen, MT, Norgaard, JV, Theil, PK, Vestergaard, M, Sejrsen, K 2006. Cell turnover and activity in mammary tissue during lactation and the dry period in dairy cows. Journal of Dairy Science 89, 46324639.CrossRefGoogle ScholarPubMed
Stelwagen, K 2001. Effect of milking frequency on mammary functioning and shape of the lactation curve. Journal of Dairy Science 84, E204E211.CrossRefGoogle Scholar
Stelwagen, K, Lacy-Hulbert, SJ 1996. Effect of milking frequency on milk somatic cell count characteristics and mammary secretory cell damage in cows. American Journal Veterinary Research 57, 902905.CrossRefGoogle ScholarPubMed
Stelwagen, K, Davis, SR, Farr, VC, Eichler, SJ, Politis, I 1994. Effect of once-daily milking and concurrent somatotropin on mammary tight junction permeability and yield of cows. Journal of Dairy Science 77, 29943001.CrossRefGoogle ScholarPubMed
Stelwagen, K, Farr, VC, McFadden, HA, Prosser, CG, Davis, SR 1997. Time course of milk accumulation-induced opening of mammary tight junctions, and blood clearance of milk components. American Journal of Physiology 273, R379R386.Google ScholarPubMed
Thompson, PA, Kadlubar, FF, Vena, SM, Hill, HL, McClure, GHY, McDaniel, LP, Ambrosone, CB 1998. Exfoliated ductal epithelial cells in human breast milk: a source of target tissue DNA for molecular epidemiologic studies of breast cancer. Cancer Epidemiology Biomarkers & Prevention 7, 3742.Google ScholarPubMed
Vandesompele, J, De, PK, Pattyn, F, Poppe, B, Van, RN, De, PA, Speleman, F 2002. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biology 3 (7): research0034.1–research0034.11.CrossRefGoogle ScholarPubMed
Wareski, P, Motyl, T, Ryniewicz, Z, Orzechowski, A, Gajkowska, B, Wojewodzka, U, Ploszaj, T 2001. Expression of apoptosis-related proteins in mammary gland of goat. Small Ruminant Research 40, 279289.CrossRefGoogle ScholarPubMed
Wilde, CJ, Knight, CH 1990. Milk yield and mammary function in goats during and after once-daily milking. Journal of Dairy Research 57, 441447.CrossRefGoogle ScholarPubMed
Zhao, FQ, Keating, AF 2007. Expression and regulation of glucose transporters in the bovine mammary gland. Journal of Dairy Science 90 (suppl. 1), E76E86.CrossRefGoogle ScholarPubMed