Parturient Cows Research Articles

Untersuchungen zur Wirkung von Kalziumlaktat bei Kühen post partum

The oral administration of calcium lactate for prophylaxis of bovine parturient paresis has been promoted for a number of years. The goal of the present study was to investigate the effect of this treatment on the serum concentrations of calcium, inorganic phosphorus and magnesium in parturient cows. Five fresh calved cows were given a drench containing calcium lactate and 5 control cows received the same drench but without calcium lactate. There were no significant differences in the serum concentrations of total and ionised calcium and magnesium between treated and control cows within 24 hours of treatment. Because the calcium lactate drench did not significantly affect calcium concentrations in the blood of fresh cows, it appears highly questionable whether the administration of calcium lactate decreases the risk of bovine parturient paresis. Based on our results, the oral administration of calcium lactate can not be recommended for prophylaxis of bovine parturient paresis.

Resistance or Susceptibility to Production-Related Metabolic Diseases (PRMDs) Can be Predicted with Fecal and Erythrocyte Isotopic Signatures

Feces and RBC isotopic signatures in close-up and parturient cows correctly classified -75% as healthy or at PRMD risk. Measuring C and N SIA as a management tool during the peripartum period may provide an economical method to identify cow resistance or susceptibility to PRMD, for retention, breeding, early treatment intervention, or culling decisions to increase profit margins.

Coxiella burnetii associated placental lesions and infection level in parturient cows

Cotyledons (n=170) from dairy cattle were analysed for Coxiella burnetii by real-time (rt) PCR targeting the IS1111a and icd genes. Positive cases (n=90) and a random selection of negative cases (n=20) were examined by histology, immunohistochemistry and, if infection level was high, by fluorescence in situ hybridisation. PCR results were compared to bulk tank milk (BTM) antibody levels. Placental infection was detected in cows from herds at all BTM antibody levels. However the likelihood of placental infection was generally higher in herds with intermediate or high BMT antibody levels than in herds with low antibody levels. Histological examination revealed a range of mostly mild cotyledonary changes; C. burnetii infection was only rarely associated with inflammation. This may explain why bovine Q fever is usually not clinically apparent. Nevertheless, infected cattle will shed C. burnetii at calving and this can occur even in herds without BTM antibodies.

Habitat selection by parturient elk (Cervus elaphus) in agricultural and forested landscapes

I examined the home range and habitat selection of 146 radio-collared female elk ( Cervus elaphus L., 1758) from 2002 to 2005 during the calving period (15 May to 24 June). I determined the proportion of home ranges of parturient cow elk during the calving period and the proportion of birthing sites of elk that were in either forested protected areas or the adjacent fragmented agriculture-dominated matrix in southwestern Manitoba, Canada. Overall, 73% of the minimum convex polygon home ranges were entirely within a protected area, 6% were only on farmland, and 21% included both. Home ranges including farmland and protected area (mean = 17.9 km2) were 3.8 times larger than those entirely inside a protected area (mean = 4.7 km2) or only on farmland (mean = 4.5 km2) (U = –2.79, P = 0.005). Female elk remaining solely in protected areas selected deciduous and mixedwood forest, marsh and fen, and water at the scale of the home range. Elk exclusively on farmland selected forage crops only. At the scale of the birthing site, females on farmland and those in protected areas selected only deciduous forest, and both types avoided agricultural cropland and marsh and fen. Identification of calving habitat will allow resource managers to manage bovine tuberculosis in the population more effectively.

Changes in Plasma Bone Metabolic Markers in Periparturient Dairy Cows

We examined fluctuations in plasma tartrate-resistant acid phosphatase isoform 5b (TRAP5b) measured using fluorometry in conjunction with those in calcium (Ca) and other bone metabolic markers from 2 weeks prepartum to 2 weeks postpartum in 7 primiparous and 18 multiparous pregnant cows. The plasma Ca concentration decreased temporarily on the day of calving in multiparous cows only. Plasma TRAP5b peaked on the day of calving in primiparous and multiparous cows and was significantly lower in multiparous cows than in primiparous cows 2 weeks before and after parturition. Plasma hydroxyproline increased 1 week postpartum in multiparous cows. Bone-specific alkaline phosphatase and osteocalcin tended to decrease after parturition in primiparous and multiparous cows. These results suggest that bone resorption increases around parturition in healthy parturient cows from the viewpoint of the TRAP5b activity.

Reciprocal expression of 17α-hydroxylase-C17,20-lyase and aromatase cytochrome P450 during bovine trophoblast differentiation: a two-cell system drives placental oestrogen synthesis

No definitive information is yet available on the steroidogenic capacity of the two morphologically distinct cell types forming the bovine trophoblast, the uninucleated trophoblast cells (UTCs) and the trophoblast giant cells (TGCs). Hence, in order to localise 17alpha-hydroxylase-C17,20-lyase (P450c17) on a cellular level and to monitor its expression as a function of gestational age, placentomes from pregnant (days 80-284; n = 19), prepartal (days 273-282; 24-36 h prior to the onset of labour; n = 3) and parturient cows (n = 5) were immunostained for P450c17 using an antiserum against the recombinant bovine enzyme. At all stages investigated, P450c17 was exclusively found in the UTCs of chorionic villi (CV), where staining was ubiquitous between days 80 and 160, but was largely restricted to primary CV and the branching sites of secondary CV between days 160 and 240. Thereafter, a distinct ubiquitous staining reoccurred in the UTCs of all CV in late pregnant, prepartal and parturient animals. Using an antiserum against human aromatase cytochrome P450 (P450arom), specific cytoplasmic staining was observed in TGCs. In placentomes from pregnant cows, staining intensity was higher in mature compared with immature TGCs and was more pronounced in the trophoblast covering big stem villi compared with the trophoblast at other sites of the villous tree. In placentomes of a parturient cow, specific staining was only found in mature TGCs that survived the normal, but substantial, prepartal decline in TGC numbers. These results clearly showed that bovine UTCs and TGCs exhibit different steroidogenic capacities, constituting a 'two-cell' organisation for oestrogen synthesis. P450c17 expression appears to be quickly down-regulated and P450arom is up-regulated when UTCs enter the TGC differentiation pathway.

Expression of Cyclooxygenase-II (COX-II) and 20α-Hydroxysteroid Dehydrogenase (20α-HSD)/Prostaglandin F-synthase (PGFS) in Bovine Placentomes: Implications for the Initiation of Parturition in Cattle

The chain of events leading to prepartal luteolysis in cattle is not yet fully understood. Prostaglandin F 2α (PGF 2α) seems to be a major factor involved. However, only little information is available about the underlying regulatory mechanisms. Consequently, the expression of cyclooxygenase-II (COX-II) and 20α-hydroxysteroid dehydrogenase (20α-HSD), an enzyme recently shown to be most likely responsible for the production of luteolytic PGF 2α in the endometrium of cyclic cows, was investigated in bovine placentomes. Immunohistochemical methods were applied to placentomes from 17 pregnant cows between days 100 and 284, from three cows during the prepartal progesterone decrease (days 273–282) and from five parturient cows. COX-II was found in uninucleated trophoblast cells (UTC) from day 100 until parturition. However, between days 100 and 235 expression was only weak to moderate, focal and mainly restricted to the chorionic plate and adjacent basal parts of chorionic stem villi. In placentomes from a 270 and a 284 day pregnant cow, in which the prepartal decline of progesterone levels had not started yet, staining had substantially increased and extended to secondary and tertiary chorionic villi. In prepartal and parturient cows strong to intense staining was present in UTC all over the villous tree. Real time RT-PCR confirmed an extensive pre- and intrapartal rise of COX-II expression in bovine placentomes with a 70–100-fold increase of COX-II-mRNA levels. 20α-HSD/PGFS was widely expressed in UTC of chorionic villi. Like COX-II it was down-regulated in UTC differentiating into trophoblast giant cells. Immunostaining pattern did not change remarkably during the period under investigation, and 20α-HSD/PGFS-mRNA levels increased only 2.6-fold in the prepartal phase. Thus, in UTC PGF 2α may be produced via COX-II and 20α-HSD/PGFS, but only COX-II may be substantially involved in the control of a putative prepartal placentomal output of luteolytic PGs, whereas 20α-HSD/PGFS seems to be expressed in a more constitutive manner.

Calving distribution of the Teshekpuk caribou herd, 1994-2003

Parturient female caribou from the Teshekpuk caribou (Rangifer tarandus granti) herd (TCH) have been observed across the western North Slope, but most cows that were seen with calves during the calving period were in the area surrounding Teshekpuk Lake. During surveys conducted between 1994 and 2003, 155 (91%) of 171 collared cows seen with calves were within an area given protected status in the 1998 Bureau of Land Management Final Integrated Activity Plan/Environmental Impact Statement (IAP/EIS). The percentage of adult collared cows seen with a calf between 1994 and 2003 has ranged from 44% to 86%, with a mean of 66%. The years with the lowest percentage of collared cows seen with calves were 1997 (50%) and 2001 (44%). In 1997 most of the herd migrated much farther south than usual, and in 2001 unusually deep, persistent snow restricted spring migration, resulting in fewer cows returning to the traditional calving area during the calving period. When snowmelt dates were earlier, calving locations were farther north. Average standardized travel rates for parturient cows were significantly greater before they had calves (7.25 km/day) than after 3.89 (km/day). Geographically, protections granted by the 1998 BLM IAP/EIS appear to adequately cover the concentrated calving grounds, allowing for variance in the annual distribution of calving cows.

Estrogen Receptor β (ERβ) Is Expressed Differently From ERα in Bovine Placentomes

The functions of placental estrogens in cattle are still poorly understood. In order to investigate a putative role as local regulators of placental growth, differentiation and functions via estrogen receptor beta (ERbeta), the expression of this ER isoform was determined in bovine placentomes obtained from cows at midgestation (days 110-150; n=7), late gestation (days 180-280; n=9), from prepartal cows (sampling immediately after the prepartal decline in maternal progesterone blood levels became obvious; n=3) and from cows at normal term (n=5) on the protein and mRNA level. By means of immunohistochemistry using a monoclonal antibody against human ERbeta (clone PPG5/10), nuclear signals were found in numerous cell types of the fetal (cotyledon) and maternal (caruncle) component of the placentomes with highest intensities in mature trophoblast giant cells (TGC), fetal and maternal vascular cells and caruncular stromal cells, in which the percentage of positive cells increased from 58.9+/-2.3 and 59.1+/-3.3 at mid- and late gestation to 65.1+/-4.9 and 69.4+/-4.2 in prepartal and parturient cows, resp. (p<0.0001). Clear staining of uninucleated trophoblast cells was only observed in prepartal and parturient animals. Conventional RT-PCR confirmed ERbeta expression in caruncles and cotyledons at all phases under investigation. With semi-quantitative real-time RT-PCR higher levels of ERbeta-specific mRNA were measured in cotyledons compared to caruncles (p<0.01). The results show that in contrast to ERalpha, which is only expressed in the caruncle, ERbeta is also extensively expressed in the cotyledon and suggest a role of ERbeta primarily in TGC differentiation and placental angiogenesis and/or vascular functions.

Effects of glucocorticoids on Fas gene expression in bovine blood neutrophils

Blood neutrophils are extremely short-lived cells that are programmed for rapid apoptosis after differentiation in bone marrow. Recently, glucocorticoids have been shown to prolong survival of human and rodent neutrophils, but the mechanisms and implications for leukocyte homeostasis and health are unclear. In this study, we investigated the effects of endogenous and exogenous glucocorticoids on Fas expression in bovine neutrophils because Fas is a major death receptor that stimulates apoptosis in circulating cells. Our study subjects were four periparturient dairy cows whose blood concentrations of cortisol peaked at calving, 15 dexamethasone-treated steers and three untreated steers whose neutrophils were exposed to dexamethasone in vitro. Fas mRNA abundance changes in collected neutrophils were monitored numerous times relative to the in vivo glucocorticoid challenges, and the relationships between these data and circulating neutrophil counts were estimated by correlation analyses. Fas mRNA and protein abundance, caspase 8 activity, and survival of neutrophils in vitro were also monitored in the presence and absence of dexamethasone. In the periparturient cows, Fas mRNA abundance in circulating neutrophils showed a sharp decrease between calving and 12 h postpartum. Based on PROC CORR analysis (SAS), this correlated negatively with blood neutrophil count (r=-0.634; P=0.0009) and serum cortisol concentration (r=-0.659; P<0.0001), but showed no relationship with serum progesterone or estradiol concentrations (P > or =0.09). Administration of dexamethasone to steers also caused a pronounced reduction in neutrophil Fas mRNA abundance that persisted for 12 h and correlated negatively with blood neutrophil count (r=-0.748; P=0.0021). In vitro, dexamethasone caused dose-dependent loss of GR proteins from the cytosol of neutrophils concurrently with Fas mRNA downregulation, which was inhibited by the glucocorticoid receptor (GR) antagonist, RU486. Dexamethasone treatment of cultured neutrophils also reduced surface Fas expression, spontaneous and sFasL-induced caspase 8 activity, and rate of apoptosis in the cells. Taken together, these in vivo and in vitro results suggest that glucocorticoids inhibit Fas expression in bovine blood neutrophils via GR activation, possibly contributing to the cells' increased longevity in culture and the pronounced neutrophilia observed in parturient cows and hormone-treated steers. We thus conclude that glucocorticoid-activated GR may change the homeostasis of circulating neutrophils, in part through its negative effects on Fas gene expression and downstream apoptosis signaling pathways.

Microarray analysis of gene expression in blood neutrophils of parturient cows.

It is well documented that blood neutrophils from parturient dairy cows do not perform as well as neutrophils from nonparturient cows in laboratory assays of adhesion, migration, or phagocytosis-induced respiratory burst. However, little is known about the possible molecular basis for parturition-induced changes in neutrophils. cDNA microarray analysis was used in the current study to explore parturition-induced changes in gene expression profiles in bovine blood neutrophils. Total RNA from isolated blood neutrophils of four parturient Holstein cows was obtained before, during, and after parturition, reverse transcribed into cDNA, and sequentially labeled with Cy3 or Cy5 dyes prior to paired hybridizations to 1,056 member bovine total leukocyte (BOTL-3) microarrays in a loop design. Resulting gene expression data were LOWESS normalized by array and analyzed using a mixed model approach. Results showed that expression profiles for 302 BOTL-3 genes were influenced by parturition. BLASTn analysis and preliminary clustering of affected genes by biological function indicated that the largest proportion (14%) of changed genes encode proteins critical to regulation of apoptosis. Independent confirmation of altered expression for 16 of these genes was achieved using quantitative real-time RT-PCR (Q-RT-PCR). A predominantly survival phenotype inferred from the microarray and Q-RT-PCR results was substantiated by monitoring apoptosis status of blood neutrophils from castrated male cattle cultured in the presence of sera from parturient cows. Thus our combined gene expression and apoptosis phenotyping results suggest that bovine parturition may induce prolonged survival in normally short-lived blood neutrophils.

Effects of extracellular Ca 2+ on phagocytosis and intracellular Ca 2+ concentrations in polymorphonuclear leukocytes of postpartum dairy cows

The aim of this study was to determine the effects of extracellular Ca 2+ concentration ([Ca 2+] e) on phagocytosis and intracellular Ca 2+ concentration ([Ca 2+] i) in bovine polymorphonuclear leukocytes (PMNs). The experiments were performed by using blood samples from parturient paretic and clinically normal parturient cows and manipulating the [Ca 2+] e in vitro. Phagocytosis by PMNs (with and without stimulation with phorbol myristate acetate and inhibition with cytochalasin B) and resting [Ca 2+] i were significantly lower in parturient paretic cows. Repletion of Ca 2+ in the extracellular media for the samples from these animals increased phagocytosis and resting [Ca 2+] i. In the blood of clinically normal parturient cows, decreasing the [Ca 2+] e decreased phagocytosis and resting [Ca 2+] i in PMNs, but increasing the [Ca 2+] e did not affect phagocytosis. These results suggest that the hypocalcemic condition of parturient paretic cows in vivo causes decreased phagocytosis and resting [Ca 2+] i in PMNs, which may partly contribute to greater susceptibility to infection.

Cervical dilatation related to uterine electromyographic activity and endocrinological changes during prostaglandin F(2alpha)-induced parturition in cows.

The temporal relationship between changes in cervical dilatation, uterine electromyographic (EMG) activity, and maternal plasma concentrations of estradiol 17beta (E(2)), progesterone (P(4)), and 13,14-dihydro-15-keto-prostaglandin-F(2alpha) (PGFM), was investigated in six parturient cows. Calving was induced with a single injection of a synthetic analogue of prostaglandin F(2alpha) (PG) on Day 274 of gestation. Cervical dilatation was measured continuously by measuring the transit time between two implanted ultrasound crystals while at the same time uterine EMG activity was measured through two silver electrodes sutured on the myometrial surface until the expulsive stage of calving had been reached. In blood samples collected at 4-h intervals, starting at the moment of PG injection, the mean plasma E(2) concentration gradually increased and was significantly elevated at 28 h after PG injection. At 4 h after PG treatment, the mean P(4) concentration had dropped significantly and continued to decrease until a value of around 1 ng/ml was reached, where it stayed until the onset of expulsion. Mean plasma PGFM concentrations increased steadily after PG injection, reaching significantly elevated concentrations at 20 h after treatment. In the five cows that delivered calves in anterior positions, uterine EMG activity, expressed as root mean square (RMS in microV), started to increase at a mean interval (+/- SD) of 13.1 +/- 3.7 h following PG treatment. The increase in EMG activity was significantly correlated with changes in plasma PGFM concentrations. In these cows, dilatation of the caudal cervix started after a mean (+/- SD) interval of 28.5 +/- 1.5 h following PG treatment and dilatation progressed at a mean (+/- SD) rate of 2.25 +/- 0.24 cm/h. In one cow with a calf in the posterior position, uterine EMG activity and dilatation started at 15.8 h and 31.8 h, respectively, after induction of calving. We conclude that a predictable sequence of physiological changes occurs around induction of calving, which allows specific timing of future studies on cellular and biochemical changes within the cervix during parturition.

The bovine placenta; a source and target of steroid hormones: observations during the second half of gestation

Apart from estrone-3-sulfate (E1S) the bovine placenta produces progesterone (P4), though the corpus luteum is the major source of P4 responsible for maintaining pregnancy. So far the biological function of placental steroids in cattle is largely unknown. However, since the local availability of free estrone (E1) in the placenta seems to be controlled by sulfatase and sulfotranferase, the hypothesis was developed that placental estrogens and P4 might act as local regulatory factors. To test for such a function placentomes from 150, 220, 240, 270 days (D) pregnant and parturient cows were screened immunohistochemically for progesterone and estrogen receptors (PR, ER). PR were found at all stages in the caruncle in stromal cells and capillary pericytes but only at parturition in arterial walls. Percentage of PR-positive caruncular stromal cells (CSC) increased ( P<0.05) from 51.8±2.6% at D150 to 58.9±1.8% at parturition. ER were detected in CSC, caruncular epithelial (CE) cells and in caruncular capillary pericytes. Mean percentage of ER-positive CSC decreased from 39.0±5.9% in pregnant cows to 17.5±8.3% at parturition ( P<0.05). In CE all cells exhibited positive signals with the exception of those immediately surrounding large primary chorionic villi. Proliferation was assessed immunohistochemically by determining the percentage of Ki67-antigen positive cells. Highest values ( P<0.001) were obtained for CE (58.0–68.3%), followed by the trophoblast (23.3–25.4%), CSC (10.6–45.3%) and the stroma of the chorionic villi (2.9–10.5%). A transient depression of proliferation in CSC between D150–270 ( P<0.05) paralleled local estrogen tissue concentrations. The results suggest that placental estrogens and P4 are important factors controlling caruncular growth, differentiation and function.

Occurrence of Estrogen Receptor α in Bovine Placentomes Throughout Mid and Late Gestation and at Parturition1

The bovine placenta produces estrogens from the first trimester until the end of its life span. However, with the exception of the immediate prepartal and intrapartal phases, in which an involvement of placental estrogens has been suggested for the preparation of parturition, their function has not been elucidated yet. To test for a role of placental estrogens as local factors regulating placental growth and differentiation, placentomes from cows that were pregnant for 150, 220, 240, and 270 days, and parturient cows (3 animals per group) were screened immunohistochemically for the expression of estrogen receptor alpha (ERalpha). Indirect immunoperoxidase staining methods were applied using primary monoclonal antibodies (pmAbs) directed against the C-terminus (AER311, HT277) or the N-terminus (AER314, 1D5) of the ERalpha molecule. Both types of pmAbs identified ERalpha in stromal cells and capillary pericytes of the maternal caruncular septae. Using pmAb 1D5, the mean percentage of ERalpha-positive caruncular stromal cells decreased from 39.0% +/- 5.9% in pregnant cows to 17.5% +/- 8.3% at parturition (P = 0.011). Only pmAb recognizing the C-terminus identified ERalpha in the caruncular epithelium, in which positive reactions were found in all cells, with the exception of areas adjacent to the chorionic plate and to major chorionic villi, where the specific signal gradually faded and occasionally disappeared. No positive reactions were observed in the fetal part of the placentomes. The expression of ERalpha in bovine placentomes was further confirmed by the detection of ERalpha-specific mRNA by reverse transcriptase-polymerase chain reaction and by Western blot analysis. The results suggest a role for placental estrogens as paracrine factors involved in the regulation of placental growth and differentiation.

Ultrasonic cervimetry to study the dilatation of the caudal cervix of the cow at parturition

The objective of this study was to investigate the temporal changes in dilatation of the caudal cervix during induced calvings ( n=5). We used ultrasound cervimetry, allowing the continuous recording of the distance between a transmitting and receiving ultrasound crystal, which were implanted opposite to each other on the caudal rim of the cervix. We started recording between 19 and 21 h after injecting a prostaglandin analogue (PG) on day 272 of gestation. A fluid-filled catheter had been introduced transcervically between the fetal membranes and the uterine wall for measurements of intra-uterine pressure (IUP). While the characteristics of calving varied widely between the five animals, it appeared possible to divide the process of dilatation into four phases. During the latent phase, which lasted until 25–43 h after PG, no net gain in dilatation occurred. We found an acceleration phase (4.3–6.8 h), in which the dilatation rate speeds up (0.49–0.84 cm/h) in three of the cows. During the phase of maximum slope (lasting 0.5–4.8 h), we measured an even higher rate (1.47–8.48 cm/h), decreasing again during the deceleration phase (rate 0.24–2.28 cm/h) in four cows. The quality of the IUP measurements precluded us from continuously investigating the relationship between cervical dilatation and uterine contractions. However, short term simultaneous recordings revealed that the cervical opening changed momentarily in the absence of IUP during the latent phase, while during the phase of maximum slope, temporary changes of dilatation coincided with uterine contractions. We concluded that the method of ultrasound cervimetry used in this study provides a valuable way to study the process of cervical dilatation in parturient cows in vivo.

Investigations on the activity of bovine placental oestrogen sulfotransferase and -sulfatase from midgestation to parturition.

To gain further information on the biological role of the high amounts of conjugated estrone (E1S) secreted by the bovine placenta, an in vitro assay system was developed to measure oestrogen sulfotransferase (OST) and oestrogen sulfatase (OS) activities in caruncular and cotyledonary homogenates and the respective subcellular fractions. Placental tissue was obtained from 150 (n = 3), 220 (n = 4), 240 (n = 3) and 270 days (n = 3) pregnant and parturient cows (n = 4). 3H-E1 and 3H-E1S served as substrates and 4-nitrophenyl sulfate potassium salt was used as a competitive inhibitor to block OS when testing for OST activity. OST-activity did not change during pregnancy and parturition and was higher (p < 0.001) in cotyledonary than in caruncular tissue with mean conversions (median) of 61.8% and 41.6% after 30 min of homogenate incubation. On a subcellular level OST-activity was clearly associated with the cytosol. OS-activity was higher (p < 0.001) in caruncular than in cotyledonary homogenates; it was constant during pregnancy (median of conversion: 88.0% and 66.4%, resp.), but was significantly decreased (p < 0.05) at parturition (median of conversion: 48.1% and 30.6%, resp.). On a subcellular level in both the cotyledon and the caruncle highest OS-activities were detected in the microsomal and the mitochondrial fractions. The decreased placentomal OS-activities in parturient cows are inconsistent with a substantial role of OS in the prepartal increase of free oestrogens. These results also suggest that bovine placental oestrogens may largely exert their action locally within the placentomes during most time of gestation as the enzymes catalyzing sulfoconjugation (i.e. inactivation of free oestrogens) and desulfation (i.e. activation of conjugated oestrogens) are expressed in close proximity to each other. In this respect the finding of oestrogen receptors in caruncular stromal cells is of particular interest.

Secretion of oxytocin in pregnant and parturient cows: corpus luteum may contribute to plasma oxytocin at term.

Plasma oxytocin (OT) concentrations were determined in 14 late-pregnant and parturient Angus-Hereford cows. Jugular and utero-ovarian veins were cannulated for simultaneous withdrawal of blood samples. Samples were collected at 10-min intervals for 6 h once weekly beginning 60-14 days before the date of expected delivery (group 1), or daily 3-7 days before the due date (group 2). In a third group, samples were collected at 15-min intervals every other day for 12 h beginning 1 wk before calving. Basal levels of OT were low, the overall mean for both veins was 0.46 +/- 0.03 microU/ml until a week before parturition, and then increased to 0.77 +/- 0.1 microU/ml (P < 0.02). Spurts of OT occurred intermittently on all days. Interpeak intervals averaged 71.0 +/- 10.7 min until Day -14, and from Day -14 to Day -1 the intervals were 44.0 +/- 5.3 min (P < 0.05). From Day -60 to Day -25 the amplitudes of OT peaks were low and similar in both veins (mean 1.37 +/- 0.1 microU/ml). From Day -14 to Day -1 the peak amplitudes were 3.6 +/- 0.4 microU/ml on average (P < 0.02). During the last 2 wk the utero-ovarian peak of OT was frequently higher than the peripheral peak. In addition, a number of spurts were observed in the utero-ovarian vein only (solo peaks). On the day of parturition during the first stage of labor, peak amplitudes had increased to 7.3 +/- 2.0 microU/ml, and the interpeak intervals had become shorter than before labor (mean 25.1 +/- 2.6 min). A large surge of OT initiated the expulsive stage of labor. Basal levels rose to 43.1 +/- 16 microU/ml and 38.7 +/- 12.6 microU/ml, and peak levels to 77.4 +/- 19.1 microU/ml and 91.6 +/- 21 microU/ml in the jugular and utero-ovarian veins, respectively. Interpeak intervals had decreased to 17.2 +/- 3.3 min (P < 0.05). Oxytocin levels remained high after delivery of the calf until the placenta was expelled. The posterior pituitary was the source of circulating OT during most of gestation and labor, but the solo peaks observed during late gestation in the utero-ovarian vein were probably of luteal origin or possibly of caruncular origin, because near term, both tissues express OT mRNA. Fetal posterior pituitary is another possible source for these peaks. Our conclusions are that during bovine pregnancy, low amplitude spurts of OT are secreted intermittently; near term, both the frequency and peak amplitude of the spurts increase; and during labor, a dramatic increase in plasma OT precedes the expulsion of the calf. The main source of OT is the posterior pituitary, but near term, a utero-ovarian source secretes additional OT into the systemic circulation.

Glucocorticoid Receptor Expression Profiles in Mononuclear Leukocytes of Periparturient Holstein Cows

Cortisol-activated glucocorticoid receptors modulate cellular responses to stress by translocating from the cytosol to the nucleus and enhancing or repressing the transcription of target genes. The functional capacity of mononuclear leukocytes is inhibited in parturient dairy cows at a time when blood cortisol concentrations are high. Because the glucocorticoid receptor is autoregulatory in many cell types, the hypothesis of the current study was that glucocorticoid receptor expression by mononuclear leukocytes is altered around parturition in association with elevated blood cortisol. If true, the glucocorticoid receptor could be involved in suppressed functions of mononuclear leukocytes in parturient cows. The objectives of this study were to determine effects of parturition on lymphocyte and monocyte glucocorticoid receptor expression and to correlate expression with serum cortisol concentrations. Objectives were achieved by using fluorescence staining and flow cytometric analyses to monitor glucocorticoid receptors in peripheral blood mononuclear leukocytes collected multiple times from 13 periparturtient test cows (eight multi- and five primiparous) and 10 midgestation control cows (five multi- and five primiparous). Serum cortisol concentrations were determined by radioimmunoassay. Based on intensity of the fluorescent glucocorticoid receptor stain, parturition caused 42 and 47% reductions in lymphocyte and monocyte glucocorticoid receptor expression, respectively, compared with mean expression in corresponding cells from control cows. When mean prepartum values were compared with nadir values at parturition in the test cows, glucocorticoid receptor expression was reduced by 67% in lymphocytes and by 54% in monocytes. Mononuclear cell expression of glucocorticoid receptors was negatively correlated with serum cortisol concentrations. Results suggest that glucocorticoid receptors are down-regulated in bovine mononuclear leukocytes in association with increased adrenal secretion of cortisol at calving. It is possible that glucocorticoid receptor down-regulation is also associated with altered phenotype or function (or both) of lymphocytes and monocytes. This possibility should be substantiated because it could explain increased disease susceptibility in periparturient dairy cows.

Immunolocalization of progesterone receptors in bovine placentomes throughout mid and late gestation and at parturition.

The corpus luteum is the main source of progesterone (P(4)) responsible for maintenance of gestation in cattle. So far it has not been possible to assign any biological role to placental P(4), which contributes only marginally and temporarily to peripheral maternal blood levels. In order to identify possible P(4) target cells within the placenta, placentomes from 150-, 220-, 240-, and 270-day-pregnant cows and from parturient cows (3 animals per group) were screened immunohistochemically for expression of the progesterone receptor (PR). During gestation, PR-positive staining was found exclusively in the nuclei of caruncular stromal cells (CSC; maternal part of the placentome) and of caruncular vascular pericytes. In placentomes from parturient cows, occasional positive nuclear staining was also observed in the walls of small caruncular arteries. The percentage of PR-positive CSC increased slightly from 51.8 +/- 2.6% on Day 150 to 56.2 +/- 5.6% at Day 270 (p < 0.05) and was 58.9 +/- 1.8% at parturition. These results suggest that in pregnant cattle, CSC are under the control of P(4) of placental rather than luteal origin. Thus, whereas luteal P(4) may regulate "coarse" systemic progestational functions in the maternal compartment in the classical hormonal manner, placental P(4) may act as a paracrine factor involved in the local regulation of caruncular growth, differentiation, and functions.