Inhibiting prolactin by cabergoline accelerates mammary gland remodeling during the early dry period in dairy cows.

The inhibition of prolactin release using cabergoline, a dopamine agonist, is an effective strategy to accelerate the changes in mammary secretion composition after drying-off. The objective of this study was to determine how cabergoline may affect mammary tissue remodeling during early involution. Holstein dairy cows were treated with either a single i.m. administration of 5.6 mg of cabergoline (Velactis, Ceva Santé Animale, Libourne, France, n = 7) or placebo (n = 7) at the time of drying-off. Mammary biopsy samples were collected 1 wk before drying-off (d -6), after 30 h of milk accumulation (d 1), and again 8 d following drying-off (d 8) to determine changes in gene expression, lactoferrin content, and cell turnover. Blood and mammary secretion samples were collected at d -6 and again at d 1, 2, 3, 4, 8, and 14 following the abrupt cessation of lactation to evaluate indicators of blood-milk barrier integrity and other markers of mammary tissue remodeling. Cabergoline induced less SLC2A1, BAX, CAPN2, and IGFBP5 mRNA expression. In contrast, cabergoline did not modify changes in cell proliferation and apoptosis. Following the cessation of lactation, changes in mammary secretion composition (Na+ and K+) and blood lactose concentrations were indicative of a loss in the blood-milk barrier function in both treatment groups. Cabergoline treatment affected only Na+ and K+ concentrations at d 1, suggesting a moderate increase in tight junction permeability. The increase in the activity of MMP9 and in mammary epithelial cell concentration in mammary secretions was greater in cabergoline-treated cows than in control cows, suggesting more mammary tissue remodeling. The increase in lactoferrin immunostaining in the mammary tissue occurred earlier for cabergoline-treated cows than for control cows, and was essentially localized in the stroma. Changes in some key markers of mammary involution suggest that cabergoline accelerates mammary gland remodeling. Thus, a single injection of cabergoline after the last milking would facilitate drying-off by enhancing mammary gland involution.

In vivo inhibition followed by exogenous supplementation demonstrates galactopoietic effects of prolactin on mammary tissue and milk production in dairy cows.

It has been previously shown that the long-term inhibition of milking-induced prolactin (PRL) release by quinagolide (QN), a dopamine agonist, reduces milk yield in dairy cows. To further demonstrate that PRL is galactopoietic in cows, we performed a short-term experiment that used PRL injections to restore the release of PRL at milking in QN-treated cows. Nine Holstein cows were assigned to treatments during three 5-d periods in a 3×3 Latin square design: 1) QN: twice-daily i.m. injections of 1mg of QN; 2) QN-PRL: twice-daily i.m. injections of 1mg of QN and twice-daily (at milking time) i.v. injections of PRL (2µg/kg body weight); and 3) control: twice-daily injections of the vehicles. Mammary epithelial cells (MEC) were purified from milk so that their viability could be assessed, and mammary biopsies were harvested for immunohistological analyses of cell proliferation using PCNA and STAT5 staining. In both milk-purified MEC and mammary tissue, the mRNA levels of milk proteins and BAX were determined using real-time reverse-transcription PCR. Daily QN injections reduced milking-induced PRL release. The area under the PRL curve was similar in the control and PRL injection treatments, but the shape was different. The QN treatment decreased milk, lactose, protein, and casein production. Injections of PRL did not restore milk yield but tended to increase milk protein yield. In mammary tissue, the percentage of STAT5-positive cells was reduced during QN but not during QN-PRL in comparison with the control treatment. The percentage of PCNA-positive cells was greater during QN-PRL injections than during the control or QN treatment and tended to be lower during QN than during the control treatment. In milk-purified MEC, κ-casein and α-lactalbumin mRNA levels were lower during QN than during the control treatment, but during QN-PRL, they were not different from the control treatment. In mammary tissue, the BAX mRNA level was lower during QN-PRL than during QN. The number of MEC exfoliated into milk was increased by QN injections but tended to be decreased by PRL injections. Injections of PRL also increased the viability of MEC harvested from milk. Although PRL injections at milking could not reverse the effect of QN treatment on milk production, their effects on cell survival and exfoliation and on gene expression suggest that the effect of QN treatment on the mammary gland is due to QN's inhibition of PRL secretion.

Effects of nutrient restriction on mammary cell turnover and mammary gland remodeling in lactating dairy cows.

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. 33.5 kg/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 α-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.

Changes in mammary secretory tissue during lactation in ovariectomized dairy cows.

In dairy animals, the milk yield (MY) changes during a lactation and is influenced by several physiological, livestock management and environmental factors. The MY produced by a mammary gland depends on synthetic activity of mammary epithelial cells (MECs) as well as MEC number and mammary secretory tissue organization. It has been suggested that ovarian steroids (estradiol and progesterone) have a negative effect on MY in lactating cows. In a previous study, we showed that the suppression of ovarian secretions by an ovariectomy improved lactation persistency in dairy cows. Here we were interested in the effects of ovariectomy on plasma estradiol and progesterone concentrations and on changes that occur in mammary secretory tissue during lactation. We demonstrated that the ovariectomy of lactating cows at the time of the lactation peak induced a rapid and dramatic drop in plasma progesterone and a smaller reduction in plasma estradiol. Interestingly, the study of the changes in mammary secretory tissue over time revealed that the improvement of MY measured in the ovariectomized cows was associated with a limited increase in estradiol receptivity in MECs, a reduced mammary tissue remodeling and reduced blood protein concentration in milk, in late lactation. These results suggest that ovarian secretions, particularly estradiol and progesterone, act to enhance processes for mammary gland involution in late-lactating dairy cows.

Effects of ovariectomy in prepubertal goats.

The objectives of this study were to determine the effects of ovariectomy on mammary gland development in prepubertal goats and to validate this model to study mammogenesis in young dairy ruminants. In this experiment, 3 months of aged goats were ovariectomized (ovx) while shammed goats played as surgery controls (sham). Thereafter, sham and ovx goats were slaughtered at 7 months of age to provide tissue for the assays. Results demonstrated that proliferation of mammary of mammary epithelial cells was significantly lower in ovariectomized goats compared to control goats. In ovx animal, epithelium structures were completely overstretched and epithelial ducts were undeveloped with limited branching whereas control animals had classical complex arborescent units with multiple round ductules and limited stroma. Concerning ERalpha (estrogen receptor alpha), PR (progesterone receptor) and P450 (aromatase) expression, results showed number of ERalpha, PR and P450 positive cells was higher in shammed goats compared to ovariectomized goats. All this results suggested that goat mammogenesis and ovarian control are similar to prepubertal heifers and that young goats are a good model to study mammary gland development in ruminants. In conclusion, we demonstrated that ovariectomy of prepubertal goats decreased proliferation of mammary epithelial cells with a profound alteration of cell adhesion molecules.