Effect of prostaglandin F2 alpha-induced luteolysis on in vivo and in vitro progesterone production by individual placentomes of cows

This study investigated placental progesterone production by bovine placentomes. Catheters were placed in the femoral artery (FA) and in the caruncular artery (CA), caruncular vein (CV) and lymphatic vessel of a prominent placentome of 13 cows at 200 d of gestation. Four of the 13 cows were given prostaglandin F2 alpha (PGF2 alpha) after surgery, and blood and lymph were collected for progesterone determination. After 24 h, progesterone was higher (P less than .01) in FA and CA plasma from control cows that FA and CA plasma from PGF2 alpha-treated cows (5.11 +/- .29 and 5.17 +/- .64 vs 1.41 +/- .08 and 1.15 +/- .08 ng/ml, respectively), but CV concentrations were similar (3.38 +/- .30 vs 2.56 +/- .24, respectively). There was a net uptake of progesterone by placentomes from control cows (P less than .01) but a net secretion in PGF2 alpha-treated cows (P less than .05). Lymph contained low progesterone concentrations regardless of treatment. Cows were slaughtered at 240 d of gestation. Placentomes were removed and perfused with pregnenolone through the maternal and fetal arteries. Fetal venous effluent contained more progesterone than maternal venous effluent (P less than .001) in both groups, and fetal venous effluent of placentomes from PGF2 alpha-treated cows contained more progesterone than that from control cows (P less than .05). Maternal and fetal components of other placentomes were cultured alone or in co-culture along with pregnenolone and (or) epostane. Fetal tissue produced more progesterone (P less than .001) than maternal tissue when each was cultured alone, but fetal tissue production declined when co-cultured with maternal tissue.(ABSTRACT TRUNCATED AT 250 WORDS)     

Structural and mechanical changes of uterine arteries during pregnancy in the pig

A dramatic 15-fold increase in uterine blood flow in pigs occurs during pregnancy in association with marked increases in uterine arterial (UA) diameter. This study was conducted to determine UA collagen and elastin content, diameter, alpha 1- and alpha 2-adrenergic receptor (AR) numbers, norepinephrine (NE) and in vitro reactivity to phenylephrine on d 0, 20, 50, 80 and 110 of pregnancy in the pig. Uterine arterial collagen content declined progressively throughout pregnancy (P less than .01), whereas the content of elastin remained constant from d 0 to d 80, then increased (P less than .05) from d 80 to d 110. The UA collagen to elastin ratio was correlated with UA diameter (r = -.69; P less than .01), which increased from 4.5 mm on d 0 to 9.0 mm on d 110. Uterine arterial alpha 1-AR numbers remained low and constant throughout pregnancy, consistent with its retained ability to contract in response to phenylephrine. Uterine arterial NE content declined (P less than .05) from d 20 to d 80 before increasing slightly to d 110. Uterine arterial alpha 2-AR numbers remained high from d 0 to d 80 before decreasing (P less than .05) to low values on d 110. These data are consistent with a reduced adrenergic neuronal control and increased elasticity of the UA during pregnancy in the pig.     

Abortifacient effects of a unique class of vasoactive lipids from Pinus ponderosa needles.

Pinus ponderosa needle (PN) ingestion by late pregnant cows results in decreased uterine blood flow, premature parturition, and retained placentae. Further, plasma from PN-fed cows increases caruncular arterial tone (i.e., induces prolonged contraction) in an isolated perfused bovine placentome. A novel class of vasoactive lipids was isolated and identified using a bovine placentome assay-guided fractionation of CH2Cl2 extracts of PN. Placentome perfusion tests indicated that 1-12-dodecanedioyl-dimyristate (14-12-14) was the most potent of the PN lipids for increasing caruncular arterial tone. Late pregnant guinea pigs (GP) were used to evaluate the abortifacient activity of these vasoactive lipids. In Study 1, on d 50 of gestation, part of the control diet was replaced with chopped PN (Diet A) or chopped PN subjected to sequential extraction with diethyl ether (Et2O; Diet B); Et2O and CH2Cl2 (Diet C); and Et2O, CH2Cl2, and methanol (Diet D). The GP on Diets A and B exhibited shorter (P<.01) gestation lengths and reduced (P<.01) pig birth weights than GP on the control diet or Diets C and D. Further, only GP on Diets A and B exhibited retained placentae. In Study 2, on d 50 of gestation, part of the control diet was replaced with chopped PN that had been subjected to exhaustive CH2Cl2 extraction and then infiltrated with either CH2Cl2 alone (Diet E), CH2Cl2 containing 14-12-14 (Diet F), or CH2Cl2 containing isocupressic acid (Diet G); then solvents were evaporated. The GP consuming Diet F had shorter (P<.05) gestation lengths and reduced (P<.05) pig birth weights than did GP consuming Diets E or G. The GP consuming Diet F also exhibited a high incidence of retained placentae. These data provide evidence that a unique class of vasoactive lipids in PN exhibit abortifacient activity in guinea pigs.     

Endocrine measurements and calving performance of Swedish red and white and Swedish Holstein dairy cattle with special respect to stillbirth

During 3 consecutive calving seasons, calving performance, placental characteristics and endocrine profiles of total 98 pregnancies of late pregnant Swedish Red and White (SRB) and Swedish Holstein (SLB) dairy heifers and cows, were investigated. Ninety-four singleton pregnancies and 4 sets of twins were recorded. In animals with singleton pregnancy, 8 stillbirths, 7 weak calves, 3 premature parturitions and 1 abortion were registered. In the SLB heifers, 19% of stillbirth (5/26) were observed, while 5% (2/42) were noted for the SRB heifers. One stillborn calf derived from the SRB cows and none was found from the SLB cows. In the heifers and cows delivering a normal living calf with unassisted parturition, the placentome thickness monitored by ultrasonography was constant towards the end of pregnancy. The numbers of foetal cotyledons varied individually between animals but in total, fewer cotyledons were found in the foetal membranes of the SRB animals than in the SLB animals (69 +/- 19) vs. (88 +/- 29) (p < 0.05). No morphological and numerical differences of the placentome thickness in animals delivering a stillborn or weak calf, compared to animals delivering a normal living calf, could be observed. In animals with unassisted parturition and without birth complications, the levels of progesterone (P4), PGF2alpha metabolite (PG-metabolite), cortisol, oestrone sulphate (E1SO4) and pregnancy associated glycoproteins (PAGs) were not different by breeds and parities. In animals carrying stillbirth, higher levels of E1SO4 were found in 3 SRB animals and 1 SLB heifer, whereas lower levels of E1SO4 were recorded in 3 SLB heifers during the last week of pregnancy, compared to the profiles found in animals with unassisted parturition. Additionally, the levels of PAGs remained low and constant in 1 SRB cow (delivering a stillborn calf), 1 SRB heifer (giving birth prematurely), 4 animals (carrying twins) and 1 aborting SRB cow. Our results show a very high rate of stillbirth in especially SLB heifers and deviating profiles of E1SO4 and PAGs in animals with impaired parturition were recorded.     

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.     

Specific effects of blood plasma from beef cows fed pine needles during late pregnancy on increasing tone of caruncular arteries in vitro.

Consumption of Ponderosa pine needles by cows during late pregnancy results in the premature delivery of viable calves. Previous observations made at necropsy suggested that vascular insufficiency in the caruncular bed was associated with this induced parturition. The present studies used the perfused caruncular arterial bed of an isolated bovine placentome to determine whether blood plasma collected from cows fed pine needles contained a vasoactive factor(s). Changes in caruncular arterial tone (i.e., long-term changes in vessel diameter) and phasic contractility (i.e., transient reductions in vessel diameter) were evaluated. Tone was quantified as the pressure exerted against a constant intraluminal flow and as the force of contraction during K(+)-stimulated depolarization of membranes. Phenylephrine, an alpha 1-adrenergic agonist, was used to stimulate a phasic contractile response. Jugular blood was collected from beef cows fed pine needles (n = 4) or a control diet (n = 4), during the last 3 d of gestation, and plasma was used for two studies. In Study 1, placentomes were perfused with decreasing dilutions (1:150 to 1:40) of plasma pooled over the last 3 d of pregnancy from cows fed pine needle or control diets. Caruncular arterial perfusion pressures and responses to K+ progressively increased (P less than .05) with decreasing dilutions of plasma from cows fed pine needles, and responses to phenylephrine remained constant. Caruncular arterial perfusion pressures and responses to K+ and phenylephrine remained constant during perfusion of plasma from cows fed the control diet. In Study 2, the changes in vasoactivity of plasma (1:60 dilution) from cows fed pine needles was evaluated during the last 3 d of pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS)     

Prostaglandin F2 alpha-induced response of the bovine ovary, oviduct (uterine tube), and uterus

Tissue strips from the ovary, (uterine tube), and oviduct, and uterus of pregnant and nonpregnant cows were tested for their contractile response to prostaglandin F2 alpha (PGF2 alpha). When 2.1 x 10(-6)M PGF2 alpha was added to the uterine strips, tension of tissues from pregnant cows increased sharply; however, tension in tissues from nonpregnant cows only increased moderately. Similar concentrations failed to elicit any response from oviductal tissues of either group. Unlike the uterus and the oviduct, the ovaries contracted slowly and irregularly. They responded with varying degrees of stimulation; ovaries from pregnant cows with brief and mild stimulation and ovaries from nonpregnant cows with slower and relatively stronger stimulation. Results indicate that the bovine ovary contracts rhythmically and that its sensitivity to PGF2 alpha decreases during pregnancy in contrast to the bovine uterus which becomes increasingly sensitive during pregnancy.     

Influence of time of initiation of a prostaglandin F2alpha protocol in dairy cows with puerperal endometritis

A field trial was conducted to elucidate the effect of the time of initiation of a repeated PGF2alpha-application in a 14 day interval for treatment of endometritis in dairy cows. On a commercial dairy farm in Brandenburg, Germany, a total of 494 dairy cows were examined by rectal palpation and adspection for signs of endometritis (vaginal discharge, enlarged uterus) between day 20 to 26 post partum (dpp). We performed two further examinations by rectal palpation and external adspection to monitor the puerperal phase (34.-40. dpp, 55.-61. dpp). All cows with symptoms of an endometritis were treated with PGF2alpha (0.15 mg R-Cloprostenol, Preloban, Intervet Deutschland GmbH Unterschleissheim) twice in a 14-day interval. In the group "Early" (n = 146) the first injection of Cloprostenol was administered at time of the 1st examination. In the group "Late" (n = 129) an identical treatment was administered in cows with endometritis, however it was started 14 days later (34.-40. dpp). The incidence of endometritis was 57.7% in the group "Early" and 53.5% in the group "Late" at the first time of examination. The 1st service conception rates for treated cows were 34% in the group "Early" vs. 37% in the group "Late". In the group "Early" differences were found in days open between treated cows with endometritis and untreated controls without symptoms of endometritis (99.1 d vs. 110.8 d, p > 0.05). In the group "Late", days open for treated (106.8 d) and untreated cows (108.0 d) were similar. The severity of endometritis influenced the percentage of cows pregnant at 200 dpp. Regarding cows with a severe endometritis (E2 and E3) the percentage of pregnant cows 200 dpp was higher in the group treated early (E2: 78.4%; E3: 80.0%) than in the group with the late initiation of the treatment (E2: 68.6%; E3: 54.5%, p < 0.05). Cows with a moderate endometritis (E1) had a similar percentage of pregnant cows (200 dpp) as the untreated cows without endometritis. It is concluded that application of PGF2alpha in the 4th and 6th week post partum in a 14 day interval in cases of severe endometritis is more effective than the application of the same treatment two weeks later.     

Plasma alpha-fetoprotein concentrations in pregnant cows exposed to Sarcocystis cruzi, Campylobacter fetus, or Aspergillus fumigatus

Bovine alpha-fetoprotein (AFP) was determined in maternal plasma, using radioimmunoassay in an attempt to detect and monitor fetal distress in pregnant cows. Plasma from pregnant cows in the 4th to 5th month of the gestation which had been exposed to Sarcocystis cruzi, Campylobacter fetus, or Aspergillus fumigatus was used. Plasma AFP concentrations were determined at intervals from before the cows were exposed until they had aborted or calved. The plasma AFP concentration of the exposed pregnant cattle remained at 6.5 +/- 5.0 mg/ml until 24 to 48 hours before abortion or parturition, when the value increased to 25.0 +/- 8.0 ng/ml. This pattern was similar for cattle exposed to each of the infective agents. Unlike in persons, rats, or monkeys, fetal-maternal transfer of AFP seems to be minimal in cows even with inflammation or necrosis of the placentome, Thus, changes in AFP concentrations in bovine plasma cannot be used as a diagnostic tool for fetal distress or fetal death.     

Endometrial expression of the insulin-like growth factor system during uterine involution in the postpartum dairy cow

Rapid uterine involution in the postpartum period of dairy cows is important to achieve a short interval to conception. Expression patterns for members of the insulin-like growth factor (IGF) family were determined by in situ hybridisation at day 14 ± 0.4 postpartum (n = 12 cows) to investigate a potential role for IGFs in modulating uterine involution. Expression in each uterine tissue region was measured as optical density units and data were analysed according to region and horn. IGF-I mRNA was localized to the sub-epithelial stroma (SES) of inter-caruncular and caruncular endometrium. Both IGF-II and IGF-1R expression was detected in the deep endometrial stroma (DES), the caruncular stroma and myometrium. IGFBP-2, IGFBP-4 and IGFBP-6 mRNAs were all localised to the SES of inter-caruncular and caruncular uterine tissue, and in the DES and caruncular stroma, with IGFBP-4 mRNA additionally expressed in myometrium. IGFBP-3 mRNA was only detectable in luminal epithelium. IGFBP-5 mRNA was found in myometrium, inter-caruncular and caruncular SES and caruncular stroma. These data support a role for IGF-I and IGF-II in the extensive tissue remodelling and repair which the postpartum uterus undergoes to return to its non-pregnant state. The differential expression of binding proteins between tissues (IGFBP-3 in epithelium, IGFBP-2, -4, -5 and -6 in stroma and IGFBP-4 and -5 in myometrium) suggest tight control of IGF activity within each compartment. Differential expression of many members of the IGF family between the significantly larger previously gravid horn and the previously non-gravid horn may relate to differences in their rate of tissue remodelling.     

Uterine blood flow during early pregnancy in ewes: interaction between the conceptus and the ovary bearing the corpus luteum

To determine if a transient increase in uterine blood flow (BF) and estradiol-17 beta (E2 beta) secretion occurs during maternal recognition of pregnancy in ewes (as previously observed for sows and cows), 40 nonpregnant (NP) ewes were assigned in equal numbers to surgery on d 9, 11, 13 or 15 postestrus (d 0 = day of estrus). For 20 NP ewes (five/day), each uterine horn (UH) was flushed with saline and uterine flushings (UF) collected. For the remaining 20 ewes, BF was determined for each UH using electromagnetic transducers, and samples of uterine arterial (UA) and uterine venous (UV) blood were obtained from each UH. After an intervening cycle, each ewe was mated, subjected to surgery on the same day postmating as during her previous nonmated cycle, and BF measurements and UA and UV samples were obtained. In addition, each UH of pregnant (P) ewes was flushed and the location of conceptuses was determined. Concentrations of E2 beta and progesterone (P4) in UA and E2 beta in UV and UF were determined by radioimmunoassay. For NP ewes, BF (ml/min) was not different for UH ipsilateral or contralateral to the ovary bearing the corpus luteum (CL), and did not differ across days, averaging 6.5 +/- .4. For P ewes, BF to UH contralateral to the ovary bearing the CL on all days and BF to UH ipsilateral to ovaries bearing CL on d 9 was similar to BF of either UH of NP ewes, averaging 6.8 +/- .6. On d 11, 13 and 15 of pregnancy, BF to UH ipsilateral to the ovary bearing CL was elevated (P less than .01) twofold (13.3 +/- .9).(ABSTRACT TRUNCATED AT 250 WORDS)     

Ontogenesis of mRNA levels and binding sites of hepatic alpha-adrenoceptors in young cattle

Catecholamines affect hepatic glucose production through (alpha- and beta2-) adrenoceptors (AR). We studied mRNA abundance and binding of hepatic alpha-AR in pre-term (P0) calves and in full-term calves at day 0 (F0), day 5 (F5) and day 159 (F159) to test the hypothesis that gene expression and numbers of hepatic alpha-AR in calves are influenced by age and associated with beta2-AR and selected traits of glucose metabolism. mRNA levels of alpha1- and alpha2-AR were measured by real time RT-PCR. alpha1- and alpha2-AR numbers (maximal binding, Bmax) were determined by saturation binding of (3H)-prazosin and (3H)-RX821002, respectively. alpha1- and alpha2-AR subtypes were evaluated by competitive binding. alpha1A-AR mRNA levels were lower in P0 than in F0, F5 and F159 and alpha(2AD)-AR mRNA levels were lower in F159 than in P0, F0 and F5, while alpha2C-AR mRNA levels increased from P0 and F0 to F5 and F159. Bmax of alpha1-AR increased from P0 to F5, then decreased in F159. Bmax of alpha2-AR decreased from F0 to F159. Bmax of alpha1-AR was positively associated with mRNA levels of alpha1A-AR (r = 0.7), Bmax of beta2-AR (r = 0.5) and negatively with hepatic glycogen content (r = -0.6). Bmax of alpha2-AR was negatively associated with Bmax of beta2-AR (r = -0.4). In conclusion, mRNA levels and binding sites of alpha1- and alpha2-AR in calves exhibited developmental changes and were negatively associated with hepatic glycogen content.     

Effect of decorin and selected glycosylation inhibitors on the adhesion of caruncular epithelial cells of pregnant cows—part I

Adhesion process ensures the formation of the appropriate connection between mother and foetus during placentation and further placental development, which determines physiological pregnancy course. Extracellular matrix of foetal membranes are a rich source of biologically active proteins, the synthesis of which is regulated by hormones. Depending on the stage of pregnancy, the protein profile of the placenta changes, thanks to which its remodelling is possible. The aim of the study was to evaluate the effect of decorin, as well as selected glycosylation inhibitors on the adhesion of caruncular epithelial cells derived from cows during pregnancy. Placental cells were isolated from healthy, pregnant (2nd and 4th month) cows after slaughter, which allowed for the establishment of 4 primary cell cultures without visible cells of fibroblast morphology. The presence of decorin in cell monolayer and cell lysates was determined by the use of immunocytochemistry and Western blotting, respectively. The viability of cells was evaluated by MTT assay. The adhesion of cells to fibronectin was measured spectrophotometrically. Protein N-glycosylation and O-glycosylation have a modulating effect on the adhesion and viability of placental cells during early–mid pregnancy. Decorin and tunicamycin were shown to have anti-adhesive properties with respect to caruncular cells of the pregnant bovine uterus.     

Expression of messenger RNA coding for 5-HT receptor, alpha and beta adrenoreceptor (subtypes) during oestrus and dioestrus in the bovine uterus

Serotoninergic and adrenergic receptors (5-HTR and AR) are involved in the regulation of uterine contractility. The objective of this study was to compare mRNA levels of 5-HTR(1A), 5-HTR(1B), 5-HTR(1D), 5-HTR(1F), 5-HTR(2A), 5-HTR(2B), 5-HTR(2C), 5-HTR(4) and alpha(1A), alpha(1B), alpha(1D), alpha(2AD), alpha(2B), alpha(2C), and beta(1), beta(2), beta(3)-AR in oestrus and dioestrus, and at three uterine locations (tip, middle and base) using quantitative real-time polymerase chain reaction. Uterine specimens consisting of endometrium and myometrium including vessels and serosa were collected from cows in oestrus (n = 10) and dioestrus (n = 15) respectively. Levels of 5-HTR and AR mRNA were expressed relative to the geometric mean of ribosomal RNA (18S), ubiquitin and glyceraldehyde phosphate dehydrogenase by the mean values of geNorm algorithm. 5-HTR(1A), 5-HTR(2C) and beta(3)-AR mRNA could not be detected in uterine tissues. The mRNA levels of 5-HTR(1F) and 5-HTR(2B) were lower (P < 0.05), but of 5-HTR(4) were higher (P < 0.05) in oestrus than in dioestrus. The mRNA levels of alpha(1A)-AR, alpha(2AD)-AR, alpha(2B)-AR were lower (P < 0.05), but of alpha(2C)-AR and beta(2)-AR were higher (P < 0.05) in oestrus than dioestrus. The mRNA levels of 5-HTR(1B) and 5-HTR(1D) (oestrus) and of alpha(2AD)-AR (dioestrus) differed among uterine locations (base > middle > tip; P < 0.05). The mRNA expression of 5-HTR and AR (subtypes) in bovine uterus was associated with cycle activity and varied according to uterine location. Additional studies on protein level will be carried out in order to elucidate the role of these receptor families on uterine contractility, which may then help to clarify clinical relevance.     

Changes in the weight, collagen concentration and content of the uterus and cervix of the ewe during pregnancy

The dry and wet weights of the uterus (caruncular and intercaruncular areas) and cervix were measured in non-pregnant (n = 5) and pregnant (n = 25) ewes post mortem; for the latter, five were obtained for each of the 5 months of gestation. The total collagen tissue content was measured in both areas of the uterus and cervix by hydroxyproline analysis and image analysis of Haematoxylin-Van Gieson stained tissue sections. Both wet and dry uterine weights increased significantly with gestational age (P < 0.001). The water content of uterine and cervical tissue remained constant, at between 83 per cent to 85 per cent and 76 per cent to 80 per cent, respectively. There was a close correlation between the two methods used to determine the collagen content (r = 0.96, P < 0.001), and between the increasing weight of the uterus during pregnancy and the total collagen content of tissues (r = 0.97, P < 0.001). At all stages, the total collagen content of the cervix [mean (SEM) 96.2 (5.4) mg g(-1)] was significantly greater (P < 0.001) than that of the caruncular mean [mean (SEM) 24.3 (1.4) mg g(-1)], and the intercaruncular areas [mean (SEM) 29.0 (1.0) mg g(-1)]. The changes in uterine and cervical weights and collagen content of the tissues were similar to those reported in other related species.     

Effect of the uterus on subnormal luteal function in anestrous beef cows

The effect of the uterus on luteal lifespan and pattern of secretion of progesterone following early weaning of calves from anestrous beef cows was studied. Calves were weaned from 15 anestrous beef cows 23 to 33 d postpartum, and cows were allotted to a control (sham surgery, n = 8) or a hysterectomy (n = 7) group, with surgery performed at weaning. Cows in the hysterectomy group were injected (im) with 25 mg prostaglandin F2 alpha (PGF2 alpha) approximately 20 d after first estrus (d 0). The interval from weaning to estrus was longer (P less than .05) for the hysterectomy group (10.4 +/- 1.6 d) than the control group (6.2 +/- .5 d). In the control group, the first estrous cycle (8.8 +/- .3 d) was shorter (P less than .01) than the second estrous cycle (20.2 +/- .5 d). Following first estrus in the hysterectomy group, cows were not detected in estrus until after injection of PGF2 alpha and did not return to estrus. From d 0 to 5, mean concentrations of plasma progesterone were similar (P greater than .05) between groups for both estrous cycles; after d 5 of estrous cycle 1, concentrations of plasma progesterone decreased in the control group. Within the hysterectomy group, the pattern of secretion of progesterone from d 0 to 16 was similar after the first and second estrus. Furthermore, there was no difference in the pattern of secretion of progesterone from d 0 to 16 between hysterectomy (first or second estrous cycles) and control (second estrous cycle) groups.(ABSTRACT TRUNCATED AT 250 WORDS)     

Alpha1-adrenoceptor-mediated negative inotropic effect caused by intracellular ionic activities in guinea-pig papillary muscle

In guinea-pig papillary muscle, phenylephrine (PE), an agonist of alpha1-adrenoceptor (alpha1-AR), led to a transient negative inotropic effect (NIE) and a subsequent sustained positive inotropic effect (PIE). To clarify the ionic mechanisms underlying the NIE, we measured the [Na+]i or [pH]i by ion-selective microelectrodes. PE produced a decrease in the intracellular Na+ concentration ([Na+]i) and an increase in intracellular pH ([pH]i). During the phase of NIE, PE produced only a (-) change of [Na+]i (Delta[Na+]i). With a decrease in extracellular Na+ or an increase in extracellular Ca2+, the PE-induced NIE was attenuated and PE produced (+)Delta[Na+]i. The PE-induced NIE and (-)Delta[Na+]i were definitely strengthened by lowering the bath temperature or increasing the stimulation frequency. 2-(2,6-di-methoxyphenoxyethyl)amino-methyl-1,4-benzidioxane HCl, an antagonist of alpha1A-AR, completely abolished the PE-induced NIE and (-)Delta[Na+]i. Phorbol 12,13-dibutyrate, an activator of protein kinase C (PKC), decreased the baseline [Na+]i and twitch force and increased the baseline [pH]i in mimicry of PE. Pretreatment with 1-5(isoquinolinesulphonyl)-2-methylpiperazine, an inhibitor of PKC, abolished the PE-induced NIE and (-)Delta[Na+]i. During pretreatment with benzamil, an inhibitor of Na+/Ca2+ exchange, we found that the PE-induced NIE and (-)Delta[Na+]i were reversibly abolished. Our results indicate the PE-induced NIE may be elicited upon the activation of Na+/Ca2+ exchange which can be attributed to the (-)Delta[Na+]i. (-)Delta[Na+]i is mediated through a PKC-dependent pathway via an activation of alpha1A-AR subtype and its effect could be strengthened remarkably at high [Na+]i and [Ca2+]i values.     

Concentration of IGF-I and estradiol-17beta in the blood plasma of pregnant cattle

Blood samples were collected from pregnant cows, heifers and also from non pregnant cows serving as controls. The determination of insulin-like-growth-factor-I (IGF-I) and estradiol-17 beta (E2) were performed immunologically. In the non pregnant cows E2 remained unchanged. IGF-I decreased around parturition and increased again commencing the 6th week of lactation. Compared to nonpregnant animals E2 but not IGF-I was slightly elevated during the first 10 weeks of pregnancy. During the 10th up to 20th week of pregnancy the mean values of IGF-I were increased as well as the ones of E2. During the late pregnancy the values of IGF-I in heifers are obviously more elevated as in lactating pregnant or non-pregnant cows; analogous high values were determined in pregnant cows only during the dry period. Before parturition even a negative correlation exists between IGF-I and E2. It is concluded that IGF-I is predominantly regulated by other factors.     

Expression of extracellular matrix metalloproteinase inducer (EMMPRIN) and its expected roles in the bovine endometrium during gestation

Extracellular matrix metalloproteinase inducer (EMMPRIN) and its induced matrix metalloproteinases (MMPs) play a crucial role in tissue remodeling during the peri-implantation period. However, the role of EMMPRIN in the bovine placenta is still unclear. We have postulated that EMMPRIN might play a regulatory role in trophoblastic cell functions during gestation by itself or through the regulation of MMP expression. In this study, EMMPRIN mRNA was detected in the bovine placentome and interplacentome throughout gestation, and its expression was significantly higher in the cotyledon during late gestation. In situ hybridization showed that EMMPRIN mRNA was expressed in the caruncular epithelium and the cotyledonary epithelium, including binucleate cells. Western blot analysis detected a band representing a protein of approximately 65 kDa in the caruncular and cotyledonary tissues, and the intensity of its expression was increased in both of these tissues during late gestation. The expression levels of MMP-2 and MMP-14 in the bovine placenta were higher during late gestation, as was observed for EMMPRIN. Therefore, EMMPRIN might regulate trophoblastic cell functions, especially those of binucleate cells, through MMP expression in the bovine placenta.