Quantification of mammary gland tissue size and composition changes after weaning in sows

The objectives of this study were to characterize the tissue compositional changes in porcine mammary glands after weaning and to determine whether administration of estradiol alters the profile of these tissue changes. Forty-five primiparous sows were assigned randomly to one of two treatment groups after weaning, control or estrogen treated. Estrogen-treated sows received twice-daily injections of estradiol-17beta (0.125 mg/kg of BW); control sows received vehicle injections. Sows were weaned at d 21 of lactation and killed on either d 0 (d of weaning; n = 5) or on d 2, 3, 4, 5, or 7 after weaning (n = 4 per treatment on each day). Teat order relative to suckling behavior was observed on the day before weaning to determine which mammary glands the piglets suckled. Suckled and non-suckled glands were identified from the teat order observation, and individual mammary glands were collected at slaughter. Mammary glands were trimmed of skin and extraneous fat pad, individually weighed, and bisected to measure cross-sectional area. The remaining half of each gland was ground and stored at -20 degrees C for chemical analyses. Frozen tissue was used for measuring tissue DNA, DM, protein, fat, and ash contents. Suckled mammary glands of sows undergo significant and dramatic changes during the initial 7 d after weaning, with significant changes occurring even by d 2 after weaning. Mean cross-sectional area of parenchymal tissue in suckled mammary glands decreased from 59.7 +/- 2.1 cm2 on the day of weaning to 26.8 +/- 2.3 cm2 by d 7 after weaning (P < 0.0001). Mammary gland wet weight decreased from 485.9 +/- 22.0 g on the day of weaning to 151.5 +/- 24.8 g by d 7 after weaning (P < 0.0001), whereas DNA decreased from 838.8 +/- 46.2 g on the day of weaning to 278.4 +/- 52.5 g by d 7 after weaning (P < 0.0001). The changes in gland wet weight and DNA during the period of mammary gland involution in the sow represent loses of over two-thirds of the parenchymal mass and nearly two-thirds of the cells that were present on the day of weaning. Estrogen treatment did not affect overall mammary involution during the first 7 d after weaning. Mammary glands that were not suckled during lactation had no further loss of parenchymal tissue during the first 7 d after weaning. Mammary gland involution in the sow is a rapid process and is probably irreversible within 2 or 3 d after weaning.     

Effect of suckling intensity on human growth hormone binding, biochemical composition and histological characteristics of ovine mammary glands

Suckling both, or only one contralateral mammary gland during 15 days postpartum was utilized to study lactogenic hormone binding to mammary microsomal membranes and quantitative mammary morphology in ewes. Binding of radiolabeled human growth hormone was specific for lactogenic hormones. Non-radiolabeled human growth hormone, ovine and bovine prolactin and human placental lactogen effectively competed with radiolabeled human growth hormone for binding sites but ovine and bovine growth hormone were completely ineffective. Specific binding of radiolabeled human growth hormone to 600 μg of membrane protein averaged 23 ± 3% in all lactating glands. Neither days postpartum nor treatment of contralateral mammary glands substantially altered hormone binding in lactating glands. Specific human growth hormone binding (6 ± 0.5%) in non-suckled glands (15 days postpartum both udder halves) was significantly lower (P<0.01) than in lactating tissue but only a moderate and variable reduction in specific binding was measured in membranes from glands non-suckled for 15 days but contralateral to a suckled gland (14 ± 4%). Specific binding was approximately doubled in assays with 600 compared with 300 μg of membrane protein and the pattern of binding among variously suckled glands was not changed by treatment of membranes with 4 M MgCl₂ prior to assay. Most secretory cells from all lactating glands had rounded, basally displaced nuclei, apical fat globules, secretory vesicles and abundant densely stained basal cytoplasm (ergastoplasm). Alveolar lumenal area was maximal (50% of tissue area) and stromal tissue area was minimal. After 15 days of non-suckling (both udder halves) mammary cells were engorged with lipid, ergastoplasm was reduced and nuclei were irregularly shaped and randomly displaced compared with lactating tissue. In addition, lumenal area was reduced and stromal tissue more evident. Lack of suckling for 5 days had little apparent effect on mammary cytology. Like lactogenic hormone binding, mammary tissue morphology was only moderately altered by 15 days of non-suckling when the remaining gland was suckled. RNA concentration was lowest (2.1 ± 0.3 mg/g) in mammary tissue from ewes in which neither gland was suckled for 15 days postpartum but non-suckling interval had no significant effect when contralateral glands were suckled. DNA concentration was not significantly influenced by suckling treatments. Relative lactogenic hormone binding closely corresponded to changes in cytological and biochemical indices of secretory cell function.     

Changes in tissue composition associated with mammary gland growth during lactation in sows.

Twenty-four primiparous sows were used to determine the extent of mammary gland growth during lactation. Litter size was set to nine or 10 pigs immediately after birth. Sows were slaughtered in groups representing d 0 (within 12 h after farrowing), 5, 10, 14, 21, and 28 of lactation. Sows were provided 17.5 Mcal ME and 65 g of lysine per day during lactation. Mammary glands were collected at slaughter and trimmed of skin and extraneous fat pad. Each gland was weighed, cut in half to measure cross-sectional area, and ground for chemical analysis. Dry matter content, dry fat-free tissue (DFFT) content, protein content, amino acids composition, ash content, and DNA content were measured. Only glands known to have been suckled were included in these data. Wet and dry tissue weight; cross-sectional area; and the amount of DFFT, tissue protein, and amino acids in each suckled mammary gland increased (P < .05) during lactation to a peak on d 21. Fat percentage of each suckled gland declined (P < .05) and the percentage of protein and DFFT increased (P < .05) as lactation progressed. These results suggest that hypertrophy occurred in the tissue during lactation. There was a linear increase in the amount and percentage of DNA during lactation (P < .05), suggesting hyperplasia of the mammary tissue. Mammary tissue growth continues in suckled glands during lactation in sows, with gland wet weight increased by 55% and total gland DNA increased by 100% between d 5 and 21 of lactation.     

Gonadotropin releasing hormone-induced secretion of luteinizing hormone during the milk-ejection reflex in the postpartum beef cow

A study was conducted to determine the effect of the milk-ejection reflex on exogenous gonadotropin releasing hormone (GnRH)-induced release of luteinizing hormone (LH) after short-term calf removal. Twenty-four postpartum multiparous beef cows were assigned randomly to groups arranged in a 2(3) factorial arrangement. Factors consisted of two levels of suckling [suckled (S) or nonsuckled (NS)], treatment with GnRH [saline (C) or 200 micrograms GnRH] and days postpartum (d 1 and 14). Dams were isolated from their calves for 4 h on d 1 and 14 postpartum. At the end of 4 h dams were reunited with their calves in S + C and S + GnRH groups, while dams of calves in NS + C and NS + GnRH groups remained separated an additional 2 h. Cows were injected iv with saline or GnRH following the 4-h isolation period, 5 min after calves had begun suckling or nuzzling the udder. Sera from jugular blood samples collected 15 min prior to the end of the 4-h isolation period, immediately prior to injection (0 h) and at 15 min intervals thereafter for 120 min were analyzed for LH. Serum concentrations of LH in control cows did not differ due to suckling or stage of the postpartum period and averaged 2.3 +/- .1 ng/ml. Pituitary response to GnRH was determined by computing the rate of LH release. Rate of LH release (ng LH.ml-1.min-1) in response to GnRH on d 14 was greater (P less than .001) than on d 1 in both suckled and nonsuckled groups (S + GnRH, 37.1 +/- 3.9 vs 18.3 +/- 5.0; NS + GnRH, 34.7 +/- 5.9 vs 14.5 +/- 1.1). However, GnRH-induced release of LH did not differ between suckled and nonsuckled cows on either d 1 or 14 postpartum. These data indicate that response of the bovine pituitary to GnRH during the postpartum period is not influenced by the act of suckling but is enhanced with time after parturition.     

Hypoglycemia alters pulsatile luteinizing hormone secretion in the postpartum beef cow

This study tested the hypothesis that the increased glucose requirement of lactation had effects that were independent of the suckling-dependent inhibition of postpartum endocrine function in beef cows. Mature Hereford cows were either suckled ad libitum and infused with saline iv (n = 9) from d 2 through 4 (d 0 = jugular catherization on d 32 +/- 3 postpartum); were nonsuckled and infused with saline from d 2 through 4 (n = 10); or were nonsuckled and infused with phlorizin (3 g/d) from d 2 through 4 (n = 10). Nonsuckled cows infused with phlorizin had lower (P less than .05) plasma concentrations of glucose and amino acid nitrogen (AAN) on d 2 compared with pre-infusion levels (d 1), but their metabolic profile returned to levels similar to the suckled cows by d 3 and 4. Nonsuckled cows infused with saline had elevated glucose and insulin and lower AAN and free fatty acids (FFA) on d 3 and 4 compared with pre-weaning (d 1) levels (P less than .05). Nonsuckled cows infused with phlorizin did not show this weaning-induced elevation in glucose and insulin. The number of luteinizing hormone (LH) pulses was not affected by treatment. However, in contrast to the large LH pulses observed in the nonsuckled cows infused with saline, both the suckled cows and the nonsuckled cows treated with phlorizin had more small and fewer large amplitude pulses (P less than .01). Treatment did not affect serum concentrations of follicle stimulating hormone, gonadotropin release in response to gonadotropin releasing hormone (25 micrograms) or the number of cows ovulating by 55 d after calving. We conclude that the increased glucose clearance caused by phlorizin infusion or lactation results in depression of LH pulse amplitude in suckled postpartum beef cows.     

Effect of nutrient intake on mammary gland growth in lactating sows

Sixty-one primiparous sows were used to determine the response of mammary gland growth to different energy and protein intakes during lactation. After birth, litter size was set to 9 or 10 pigs. Sows were slaughtered at selected times up to 30 d of lactation. Individual sows were fed one of four diets that were combinations of different amounts of energy and protein (3.0 Mcal ME and 8.0 g lysine/kg diet; 3.0 Mcal ME and 16.2 g lysine/kg diet; 3.5 Mcal ME and 6.4 g lysine/kg diet; or 3.5 Mcal ME and 13.0 g lysine/kg diet). Mammary glands were collected at slaughter and trimmed of skin and the extraneous fat pad. Each gland was weighed, cut in half to measure cross-sectional area, ground, and stored at -20 degrees C for chemical analysis. Frozen, ground tissue was used to determine dry matter, dry fat-free tissue (DFFT), total tissue protein, ash, and DNA content. Only glands known to have been suckled were included in this data set. Response surface regression was used for statistical analysis. The percentage of protein, fat, ash, and DNA in each suckled mammary gland was affected only by total energy intake (P<.05). The percentage of dry tissue and fat decreased as the total energy consumed during lactation increased, whereas the percentage of protein and DFFT increased as total energy intake increased. There were quadratic effects (P<.05) of both total energy and protein intake on wet weight, dry weight, protein amount, DFFT amount, and DNA amount of each suckled mammary gland during lactation. This study shows that mammary gland growth is affected by nutrient intake during lactation. The weight of suckled mammary glands and the amount of mammary tissue protein, DFFT, and total DNA were maximal on d 27.5 of lactation when sows had consumed an average of 16.9 Mcal of ME and 55 g of lysine per day during lactation. Provision of adequate amounts of nutrients to sows during lactation is important for achieving maximal growth of mammary glands and maximal milk production.     

The regression of unsuckled mammary glands during lactation in sows: the influence of lactation stage, dietary nutrients, and litter size.

During lactation in the sow, mammary glands that are not regularly suckled undergo regression. This study characterizes the regression of unsuckled mammary glands and how that regression is affected by dietary nutrients and litter size. Sixty-nine primiparous sows were fed one of four diets containing combinations of two protein levels (32 or 65 g lysine/d) and two energy levels (12 or 17.5 Mcal ME/d) during lactation. Litter size was adjusted to 10. Sows were killed on d 0, 5, 10, 14, 21, or 28 of lactation. In another experiment, twenty-eight primiparous sows were allotted to have different litter sizes and were killed on d 21 of lactation. The day before slaughter, teat order of each litter was observed. After death, mammary glands were removed and dissected. Skin and extraneous fat pads were removed from the mammary glands and individual glands were separated. Each gland was weighed, cut in half to measure cross-sectional area, and ground for chemical analysis. The amounts of dry tissue, protein, fat, ash, and DNA were measured. Only glands observed to be unsuckled were included in the results. Regression of unsuckled mammary glands occurred rapidly during the first 7 to 10 d of lactation, as indicated by a decline in wet weight, dry weight, protein, fat, DNA, and cross-sectional area. The rate of regression was slowed after the early lactation period. The rate of regression of unsuckled glands was affected by dietary nutrient levels. Dietary energy level affected (P < 0.05) the decline in wet and dry weights, protein, fat and DNA content, and cross-sectional area, whereas dietary protein level affected (P < 0.05) the decline in dry weight and fat content. At d 5 of lactation, the wet weight of unsuckled mammary glands in sows fed the high-energy high-protein diet was 91% greater (P < 0.05) than in sows fed the low-energy low-protein diet. Effects of litter size on size and composition of unsuckled glands were not significant by d 21 of lactation. Unsuckled mammary glands regress rapidly during early lactation, and the rate of regression is affected by dietary nutrient intake.     

Morphologic changes in the bovine mammary gland during involution and lactogenesis

Morphologic changes developing during bovine mammary involution were examined. Quarter biopsy specimens were obtained weekly from 5 cows beginning the day milking was discontinued through parturition. Light and electron microscopic examination of mammary tissue indicated a gradual reduction in synthetic and secretory activity of alveolar epithelium as involution progressed. Light microscopic morphologic analysis revealed increases in stroma and nonactive secretory epithelium, with concomitant decreases in epithelium, lumen, and fully active secretory epithelium during the first 2 weeks of involution. Electron microscopic analysis of alveolar epithelium revealed decreased number of organelles associated with milk synthesis and secretion during this time. These changes reversed gradually beginning 2 weeks before parturition, and by the time of calving, cell structure was typical of lactating glands. Tissue from infected quarters had less synthetic and secretory ability as indicated by significantly higher percentages of stroma and nonactive cells, but lower percentages of lumen and moderately active cells, compared with uninfected quarters. Infected quarters also had more leukocytes infiltrating the epithelium, lumen, and stroma, compared with uninfected quarters. Microscopic examination of macrophages and neutrophils suggested these cells removed milk components and cellular debris during involution. Large numbers of plasma cells, with distended cisternae of rough endoplasmic reticulum, suggested local antibody production during the periparturient period.     

Suckling inhibits release of luteinizing hormone-releasing hormone from the bovine median eminence following ovariectomy

The effect of suckling on depletion of hypothalamic LHRH from the median eminence (ME) following ovariectomy (OVX) was determined in cattle. Multiparous, postpartum Holstein cows were assigned randomly to three groups: intact, nonsuckled (INT, n = 4); ovariectomized (3 to 5 d after parturition), nonsuckled (OVX, n = 4); and ovariectomized (3 to 5 d after parturition) and suckled by three calves (OVX-S, n = 5). Blood samples were collected at three periods (1 to 7 d before parturition and 3 to 5 d and 31 to 37 d after parturition) to determine plasma LH concentration. At 31 to 37 d after parturition, all cows were slaughtered and each ME was collected and mid-sagitally sectioned. The left half of each ME was used to determine content and concentration of LHRH. Concentrations of LH and LHRH were determined by RIA. Plasma LH concentration was similar among the three groups at 1 to 7 d before parturition and 3 to 5 d after parturition; however, at 31 to 37 d after parturition, OVX cows had a greater (P less than .05) concentration of LH (2.25 +/- .64 ng/ml) than either INT (.47 +/- .10 ng/ml) or OVX-S (.92 +/- .14 ng/ml) cows. Content of LHRH in the ME of INT (80.12 +/- 15.0 ng) and OVX-S 109.8 +/- 16.4 ng) cows was similar but was greater (P less than .05) than that in OVX cows (48.95 +/- 5.9 ng).(ABSTRACT TRUNCATED AT 250 WORDS)     

Quantitation of calmodulin in the mammary gland of the lactating sow

It has been suggested that calmodulin, a calcium-binding protein, has a functional role during milk secretion. High levels of calmodulin are present during lactation in rat mammary glands and a substantial increase has been observed in the bovine mammary gland prior to parturition. In the sow, regressed glands involute while suckled glands remain highly active even though they are under the same hormonal influence. In this study, tissue samples were taken from suckled and regressed glands of the same sow at both peak and late lactation. Calmodulin and total protein were measured in tissue homogenate supernatants. Residual milk was apparent in regressed glands during mid lactation but not in the same glands by late lactation. Calmodulin levels in tissue were the same for both suckled and regressed glands. There was a slight but non-significant increase in the tissue calmodulin level from peak to late lactation. Protein levels declined significantly from mid to the late stage of lactation. There was no change in protein level between the suckled and regressed glands. Calmodulin may be responsible for casein phosphorylation and/or the mediation of prolactin action on the gland. The precise regulatory mechanisms relating hormonal control to calmodulin levels during lactation need further investigation.     

Anovulation in postpartum suckled beef cows. I. Associations among size and numbers of ovarian follicles, uterine involution, and hormones in serum and follicular fluid

Changes in sizes and numbers of ovarian antral follicles, uterine size and weight, serum hormones, and frequency and duration of suckling were examined during the postpartum anovulatory period in primiparous, suckled beef cows. Twenty-one anovulatory, suckled cows (n = 4 to 6/d) were slaughtered on d 7, 14, 28 and 42 to 56 after parturition. In addition, a total of 11 postpartum cows that had begun cyclic activity were slaughtered on d 28, 42 or 56. Blood was collected at 10-min intervals for 6 h 1 d before slaughter for measurement of prolactin, cortisol and progesterone in serum. Numbers of medium (4.0 to 7.9 mm) follicles increased fourfold (P less than .05) between d 7 and 42 to 56 in anovulatory cows, whereas numbers of small (1.0 to 3.9 mm) and large (greater than or equal to 8.0 mm) follicles did not change (P greater than .10). Uterine involution was complete by d 28. In anovulatory cows, a higher (P less than .05) proportion of largest (but not second-largest) follicles was opposite the ovary containing the corpus albicans from pregnancy (CAP). In addition, 90% of these largest follicles opposite the CAP had concentrations of estradiol greater than progesterone. In cyclic cows, however, first ovulations occurred with equal frequency on either ovary. Concentrations of prolactin or cortisol in serum or duration of suckling were not associated with changes in uterine or ovarian measurements. In conclusion, growth and function of the largest (but not second-largest) follicle were reduced when located on the ovary containing the CAP.(ABSTRACT TRUNCATED AT 250 WORDS)     

Morphometric study of experimentally induced Staphylococcus bovis mastitis in the cow

Comparisons were made of bovine mammary tissue at 12 and 24 hours after 5,000 colony-forming units of Staphylococcus aureus 305 was injected into the mammary parenchyma with control tissue from the contralateral udder half. A mild form of staphylococcal mastitis was induced routinely. Generally, S aureus was found in udder tissue removed centrally from the site of injection. Varied degrees of tissue damage were found in infected quarters from alveolar epithelium without damage (similar to control) to extremely swollen epithelium which were typical of cells undergoing disruption. Infected tissues frequently displayed areas of swollen stroma and areas of diminished alveolar lumens. Cellular debris was often found in lumens, and neutrophils frequently filled the alveolar lumen in areas of greatest damage. Thus, a parenchymal injection of bacteria induced early forms of staphylococcal mastitis at the alveolar level.     

Influence of the ovary and suckling on luteinizing hormone response to naloxone in postpartum beef cows

The influence of the suckling stimulus and ovarian secretions on LH response to naloxone was studied in 16 postpartum anestrous beef cows that were assigned randomly to one of four groups (n = 4/group): intact suckled (IS), intact nonsuckled (IN), ovariectomized suckled (OS) or ovariectomized nonsuckled (ON). Ovariectomy (OS + ON) and calf removal (IN + ON) were performed on d 2, 3 or 4 after parturition. Jugular venous blood was collected at 15-min intervals for 4 h before and 4 h after administration of naloxone (1 mg/kg BW, i.v.) on d 14 and d 28 after parturition. Gonadotropin-releasing hormone (5 micrograms, i.v.) was given 3 h after naloxone. Both IN and OS increased (P less than .05) mean pretreatment LH above IS values (mean +/- SE, ng/ml; IS 1.6 +/- .1 vs IN 2.5 +/- .3 and OS 2.7 +/- .4; P less than .01), whereas ON increased (P less than .01) LH (3.7 +/- .3 ng/ml) even further. Mean LH increased (P less than .05) after naloxone administration in all treatment groups. However, magnitude of this response was variable and dependent on ovarian status. Amplitude of the naloxone-induced LH response was greater (P less than .05) for ovariectomized (5.9 +/- 1.1 ng/ml) than for intact groups (2.7 +/- .5 ng/ml). Gonadotropin-releasing hormone increased mean LH concentrations in all groups. We suggest that ovarian secretions and the suckling stimulus contribute to endogenous opioid inhibition of LH during the postpartum interval.(ABSTRACT TRUNCATED AT 250 WORDS)     

Suckling as a regulator of postpartum rebreeding in cattle: a review

An extended and variable period of anovulation and anestrus occurs in suckled beef cows following parturition. This imposes both biological and economic constraints on the efficiency of beef production. A large body of knowledge has linked these phenomena to an attenuation of neuroendocrine signals that subserve gonadal function. During late gestation, high concentrations of placental estrogen inhibit the synthesis of LH, and pituitary stores of LH are depleted at parturition. Maximum pituitary stores of LH, releasable pools of LH and the ability of hypothalamic centers to respond to positive feedback effects of estradiol occur within 3 to 4 wk after calving in suckled cows. However, the requisite pattern of pulsatile LH secretion, which accompanies similar changes in nonsuckled cows within 2 to 3 wk after calving, occurs in only 30 to 50% of suckled animals. The period of acyclicity that continues in the remainder is exacerbated by poor body condition and may persist in some females for periods exceeding 100 d. Utilizing data from the cow, as well as other species, a model for suckling-induced inhibition of pulsatile LH release is proposed. The biological and economic impact of current and future techniques for controlling suckling-induced anestrus also is considered, including a conceptual analysis of nonbiological limitations.     

Assessing dairy potential of western white-faced ewes

Two experiments were conducted to examine the milk producing ability of Western White-Faced sheep and to identify traits that correlate well with milk production. In Exp. 1, 31 Targhee ewes were milked and five samples were taken during 107-d lactations in which the ewes nursed twin lambs. Milk yield and composition, lamb weights, ewe weights, wool growth, and udder size also were measured. In Exp. 2, 24 ewes (Rambouillet x Finn-Dorset) were separated from their lambs at 7 wk and milked twice per day for eight more weeks, during which milk yield and composition, feed consumption, udder width, and ewe weights were measured. Results from Exp. 1 showed that lamb 30-d weights, ewe weights at breeding time, and udder width at peak lactation were highly correlated with suckled milk yield (r = .81, .75 and .66, respectively). Results from Exp. 2 indicated that lamb weights and ewe weights were not useful for predicting milk yield in dairy ewes, but feed intake and udder width were (r = .74 and .86, respectively). Single-day milk yield measurements were excellent estimators of total lactation yield in both experiments. Milk yields averaged 1,714 g/d in the suckled ewes and 477 g/d in the dairy ewes.     

Synchronized estrus and fertility of beef cows after weaning calves for short intervals

In Exp. 1, the objective was to determine if interval of separating calves from cows (24 or 48 h) immediately before insemination affects detection and precision of estrus and pregnancy rates of lactating beef cows implanted with norgestomet. Separation of calves from cows for 24 h (n = 418) lengthened intervals to estrus, did not affect precision of estrus, reduced success of detecting estrus and lowered pregnancy rates relative to positive controls (48 h separation, n = 508). Cows with poor body condition, and not suckled for 24 h, conceived at lower rates than cows with similar condition that were not suckled for 48 h. Adverse effects of separation for only 24 h on fertility are apparently due to inadequate intervals between estrus and insemination at 48 h after removing implants. In Exp. 2, the objective was to determine effects of separating calves from cows for 48 h immediately before insemination on detection and precision of estrus and on pregnancy rate of ovulatory lactating beef cows injected twice with prostaglandin F2 alpha (PGF2 alpha). Weaning increased detection of estrus but overall pregnancy rates did not differ between suckled (n = 256) and nonsuckled (n = 221) cows. But, weaning calves improved pregnancy rates of young (2 to 3 yr) cows and reduced fertility among middle (4 to 6 yr)-aged cows. Increased pregnancy rates after weaning calves for 48 h are due largely to greater detection of estrus and inseminating more cows.(ABSTRACT TRUNCATED AT 250 WORDS)     

Involution and redevelopment of the mammary gland in goats. Histological-histochemical studies after a lactation time of 8-10 months

7 chamois-coloured mountain goats were used to investigate histologically and histochemically the processes of involution and redevelopment of the mammary gland after a lactation period of 8 to 10 months. Tissue specimens were obtained by incision biopsy at drying-off two months prepartum and afterwards in intervals of 8-16 days up to parturition. The findings of this investigation were compared with results of two former investigations, in which involution and proliferation of the gland were studied separately, thus precluding an overlapping of the two processes. At drying-off, after an 8 to 10 month-lactation, sections indicative of active lactation occurred concomitantly with sections at various stages of involution, as well as early stages of redeveloping alveoli. At 16 days post drying-off, the sections indicating lactation at first examination were in a stage of maximum involution and transformation. Thus, the time required for involution was reduced by half compared to drying-off at peak lactation. At 32 days post drying-off, stages of proliferation predominated with only a few involuting glands. Specimens obtained thereafter contained only redeveloping glands. The histological and histochemical differentiation between areas of involution and those of proliferation may present difficulties during the mid-portion of the dry period. There appears to be an association between the length of the time interval from drying-off to parturition and the rate of tissue transformation in the caprine mammary gland; the rate is increased when the duration of the dry period is reduced.     

Effect of continuous milking and prostaglandin E2 on milk production and mammary epithelial cell turnover, ultrastructure, and gene expression

Mammary epithelial cell (MEC) growth is reduced in continuously milked (CM) mammary glands, and administration of a mammogenic compound such as prostaglandin E(2) (PGE(2)) at parturition might improve MEC growth in CM tissue. The objectives were to 1) compare MEC turnover, ultrastructure, and gene expression in CM and involuting mammary tissue, and 2) evaluate the effects of CM and intramammary infusion of PGE(2) on early lactation MEC turnover, ultrastructure, mammary gene expression, milk yield, and composition. First- and second-lactation cows (n = 8) were used in a half-udder model, in which one-half was dry for 60 d (CTL) and the other was CM. Udder halves (n = 16) were assigned to a postpartum (PP) treatment of PGE(2) (+PGE(2); 875 mug/10 mL of medium-chain triglyceride oil) or no PGE(2) (-PGE(2)) treatment at parturition and at 72 h PP. Biopsies of CM and CTL quarters were obtained during milk stasis (MS) of the CTL half at 3 and 7 d after dry-off of the CTL half (3d-MS; 7d-MS) and postpartum (PP) at 2 and 4 d (2d-PP; 4d-PP). Milk yield was reduced (P < 0.01) in CM udder halves compared with CTL halves (13.2 vs. 22.1 kg/d), but reductions were less in second-lactation cows. The apoptotic index was greater (P < 0.05) in CTL glands than in CM glands (3d-MS, 0.52 vs. 0.11% and 7d-MS, 0.24 vs. 0.12, respectively). Proliferation of MEC was unchanged at 3d-MS, but was increased (P = 0.01) in CTL halves at 7d-MS compared with CM halves (3.10 vs. 0.93%). At 2d-PP, MEC proliferation was increased (P = 0.05) in CM halves compared with CTL halves (1.3 vs. 0.6%), but was unaffected by PGE(2) (P > 0.2). Apoptosis was elevated in early lactation regardless of treatment. Ultrastructure was unchanged by dry period length or PGE(2). In prepartum tissue, involution in CTL halves increased (P < 0.05) the expression of the proapoptotic genes Bcl-2-associated x protein (bax) and IGFBP5 and decreased (P < 0.05) alpha-lactalbumin expression compared with CM tissue. In PP mammary tissue, CTL halves expressed greater (P < 0.05) levels of ATP-binding cassette 1 (ABC1) and IGFBP5. Treatment with PGE(2) did not alter (P > 0.1) gene expression. The results confirm that CM reduced milk yield of cows with a mammary growth requirement. Reduced MEC turnover and milk yield were not alleviated by IMI of PGE(2), which indicates that peripartum PGE(2) concentrations in CM glands are not limiting mammary growth or milk synthesis.