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20. Use of somatic cells from goat milk for dynamic studies of gene expression in the mammary gland

Author

Boutinaud, M., Rulquin, H., Keisler, D.H., Djiane, J., and Jammes, H.

Subjects
Somatic cells -- Research, Goat's milk -- Research, Zoology and wildlife conservation
Abstract

Somatic cells are present in the milk throughout lactation and consist of leukocytes and epithelial cells exfoliated from the mammary epithelium. Our objective was to determine the efficacy of using somatic cells from goat milk for dynamic studies of gene expression in the mammary gland. Over a 4-wk interval, cells were isolated from daily morning milk samples and samples taken 30 min after milking. They were characterized by direct cell counts and by flow cytometry analysis after immunostaining with antibodies directed against cytokeratin and CD45, a common leukocyte antigen. Epithelial cell counts within the morning milk ranged from 15 to 45% of total milk somatic cells. After-milking samples contained twice as many cells as did morning milk samples. The RNA was extracted from the somatic cells of both types of milk samples with equivalent efficiency (a mean of 1.2 [micro]g RNA/mL of milk). Four mRNA variants of the [alpha]-S1 casein gene were detected by Northern blot analysis and the amount of each mRNA in milk cells was related to protein concentration in milk. The comparison between mRNA from the mammary gland and from congruently collected milk cells showed that relative amounts of mRNA for each milk-protein ([alpha]-S1 and [kappa]-casein and [alpha]-lactalbumin) were conserved. In a third experiment, daily milk cell RNA preparations were extracted to assess the effect of growth hormone (GH) on mammary gene expression; four goats were separated into two groups in order to perform a switch-back design consisting of three treatment weeks: Control, GH-Control or GH-Control-GH. In this study, treatment of goats with GH increased milk yields by 5%. Throughout the control and GH treatments, the expression of the three milk-protein genes studied were highly and significantly correlated (r = 0.949 and r = 0.958, P < 0.001 for, respectively, [alpha]-S1 and [kappa]-casein and for [alpha]-S1 casein and [alpha]-lactalbumin). During GH treatment, the three milk-protein mRNA abundances increased with the same pattern. In conclusion, the opportunity to use milk somatic cells for RNA preparation and analysis provides a significant improvement over the use of biopsy samples in assessing gene activity in the mammary gland and allows easy and repetitive sampling without damaging mammary tissue. Furthermore, we propose that this method could be used to investigate the transcriptional status of the mammary gland of an animal in relation to its genotype, nutritional and pathologic status, and under influence by hormonal factors. Key Words: Goats, Mammary Tissue, Messenger RNA, Somatotropin

Published
2002

34. The present situation and perspectives on milking and milk production of dairy sheep in Europe, the Middle East and North America

Author

Marnet, Pierre-Guy, Lagriffoul, G., Morin, E., Allain, C., Larroque, H., Rupp, R., Astruc, J.M., Dessauge, F., Boutinaud, M., Lollivier, Vanessa, Duvallon, O., Aurel, M.R., Autran, P., Production du lait (PL), Institut National de la Recherche Agronomique (INRA)-AGROCAMPUS OUEST, and Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)

Subjects
fluids and secretions, [SDV.SA.SPA]Life Sciences [q-bio]/Agricultural sciences/Animal production studies, food and beverages
Abstract

The present situation and perspectives on milking and milk production of dairy sheep in Europe, the Middle East and North America

Published
2010

35. Effects of nutrient restriction on mammary cell turnover in lactating dairy cows

Author

Dessauge, F., Lollivier, V., Laurence FINOT, Wiart, S., Cutullic, E., Disenhaus, C., Barbey, S., Lemosquet, S., Boutinaud, M., Production du lait (PL), Institut National de la Recherche Agronomique (INRA)-AGROCAMPUS OUEST, Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro), UE : Domaine Expérimental du Pin 0326, and Institut National de la Recherche Agronomique (INRA)

Subjects
fluids and secretions, [SDV]Life Sciences [q-bio], [SDV.SA.SPA]Life Sciences [q-bio]/Agricultural sciences/Animal production studies, food and beverages, [INFO]Computer Science [cs]
Abstract

15 crossbred dairy cows divided into control (basal diet; n=7) and restricted diet (60% grass silage and 40% hay; n=8) groups. Milk yield and composition were determined, and mammary glands were removed and weighed after 11 weeks of lactation. Results revealed that milk yield was lower in restricted diet group from calving to slaughter. Restricted feeding decreased milk protein content and milk lactose content with similar milk fat composition. Mammary glands of restricted diet group cows were lighter than control. Apoptosis in the mammary gland was higher in the restricted diet animals. It is concluded that lower milk yield induced by nutrient restriction in lactating dairy cows is partly due to the lower number of mammary cells.

Published
2010

36. Milking-induced prolactin release has proliferative and survival roles on mammary epithelial cells in dairy cows

Author

Boutinaud, M., Angulo Arizala, J., Bruckmaier, R.M., Lacasse, P., Lollivier, Vanessa, Production du lait (PL), Institut National de la Recherche Agronomique (INRA)-AGROCAMPUS OUEST, and Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)

Subjects
endocrine system, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, food and beverages, hormones, hormone substitutes, and hormone antagonists
Abstract

Milking-induced prolactin release has proliferative and survival roles on mammary epithelial cells in dairy cows

Published
2010

37. Effects of nutrient restriction on mammary cell activity and hormonal statement in lactating dairy cows

Author

Dessauge, F., Lollivier, V., Cutullic, E., Portanguen, J., Disenhaus, C., Barbey, S., Ponchon, B., Boutinaud, M., Production du lait (PL), Institut National de la Recherche Agronomique (INRA)-AGROCAMPUS OUEST, and Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)

Subjects
[SDV.SA.SPA]Life Sciences [q-bio]/Agricultural sciences/Animal production studies, food and beverages
Abstract

The aim of this study was to investigate the effects of a severe nutrient restriction on mammary tissue morphology and remodeling, mammary epithelial cell (MEC) turnover and activity, and hormonal status in lactating dairy cows. We used 16 Holstein × Normande crossbred dairy cows, divided into 2 groups submitted to different feeding levels (basal and restricted) from 2 wk before calving to wk 11 postpartum. Restricted-diet cows had lower 11-wk average daily milk yield from calving to slaughter than did basal-diet cows (20.5 vs. 33.5kg/d). Feed restriction decreased milk fat, protein, and lactose yields. Restriction also led to lower plasma insulin-like growth factor 1 and higher growth hormone concentrations. Restricted-diet cows had lighter mammary glands than did basal-diet cows. The total amount of DNA in the mammary gland and the size of the mammary acini were smaller in the restricted-diet group. Feed restriction had no significant effect on MEC proliferation at the time of slaughter but led to a higher level of apoptosis in the mammary gland. Gelatin zymography highlighted remodeling of the mammary extracellular matrix in restricted-diet cows. Udders from restricted-diet cows showed lower transcript expression of a-lactalbumin and kappa-casein. In conclusion, nutrient restriction resulted in lower milk yield in lactating dairy cows, partly due to modulation of MEC activity and a lower number of mammary cells. An association was found between feed restriction-induced changes in the growth hormone-insulin-like growth factor-1 axis and mammary epithelial cell dynamics.

Published
2010

38. Prolactin-release inhibition and once daily milking affect mammary cell activity at a transcriptional level

Author

Boutinaud, M., Lollivier, Vanessa, Bruckmaier, R.M., Lacasse, P., Production du lait (PL), Institut National de la Recherche Agronomique (INRA)-AGROCAMPUS OUEST, and Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)

Subjects
fluids and secretions, [SDV.SA.SPA]Life Sciences [q-bio]/Agricultural sciences/Animal production studies, food and beverages
Abstract

Inhibiting milking-induced prolactin (PRL) release and reducing milking frequency reduce the quantity of milk produced by dairy cows. Nevertheless, the mechanisms by which these two parameters modulate milk yield are not clear. To assess the regulation of the cell activity, by PRL and milking frequencies, five Holstein cows in early lactation received daily i.m. injection of 1 mg of Quinagolide, a suppressor of PRL release, for 9 weeks. Four control cows received the vehicle (water). During the last week of treatment, one half-gland was milked once and the other twice daily. Mammary biopsies were taken after 4 and 8 weeks of quinagolide treatment and epithelial cells were isolated from milk of both halvesudders in both groups of animals during the differential milking. Epithelial cells were prepared after milk centrifugation and by purification using an anticytoketatin antibody bound to magnetic beads. RNA was extracted from mammary biopsies and milk purified epithelial cells. Milk protein mRNA expression were analysed by quantitative real time reverse transcription PCR. Quinagolide treatment induced a faster decline in milk production (- 5.3 kg/d compared to control during the last 4 weeks of treatments, P 0.15). Levels of kappa casein (P

Published
2009

39. Hormone and growth factor regulation of tissue remodeling in the mammary gland

Author

Flint, D., Allan, G., Beattie, J., Travers, M., Barber, M., Andreas Kolb, Whitelaw, C., Boutinaud, M., Binart, N., Kelly, P., Research Institute, Roslin Institute, Production du lait (PL), Institut National de la Recherche Agronomique (INRA)-AGROCAMPUS OUEST, Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro), and Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
PROTEASES, [SDV.SA]Life Sciences [q-bio]/Agricultural sciences, OBESITY, IGF, ComputingMilieux_MISCELLANEOUS
Abstract

International audience

Published
2005

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