Down-regulation of oxytocin-induced cyclooxygenase-2 and prostaglandin F synthase expression by interferon-tau in bovine endometrial cells.
Oxytocin (OT) is responsible for the episodic release of luteolytic prostaglandin (PG) F2alpha from the uterus in ruminants. The attenuation of OT-stimulated uterine PGF2alpha secretion by interferon-tau (IFN-tau) is essential for prevention of luteolysis during pregnancy in cows. To better understand the mechanisms involved, the effect of recombinant bovine IFN-tau (rbIFN-tau) on OT-induced PG production and cyclooxygenase-2 (COX-2) and PGF synthase (PGFS) expression in cultured endometrial epithelial cells was investigated. Cells were obtained from cows at Days 1-3 of the estrous cycle and cultured to confluence in RPMI medium supplemented with 5% steroid-free fetal calf serum. The cells were then incubated in the presence or absence of either 100 ng/ml OT or OT+100 ng/ml rbIFN-tau for 3, 6, 12, and 24 h. OT significantly increased PGF2alpha and PGE2 secretion at all time points (p < 0.01), while rbIFN-tau inhibited the OT-induced PG production and reduced OT receptor binding in a time-dependent manner. OT increased the steady-state level of COX-2 mRNA, measured by Northern blot, which was maximal at 3 h (9-fold increase) and then decreased with time (p < 0.01). OT also caused an increase in COX-2 protein, which peaked at 12 h (11-fold increase), as measured by Western blot. Addition of rbIFN-tau suppressed the induction of COX-2 mRNA (89%, p < 0.01) and COX-2 protein (50%, p < 0.01) by OT. OT also increased PGFS mRNA, and this stimulation was attenuated by rbIFN-tau (p < 0.01). To ensure that the decrease in COX-2 was not solely due to down-regulation of the OT receptor, cells were stimulated with a phorbol ester (phorbol 12-myristate 13-acetate; PMA) in the presence and absence of rbIFN-tau. The results showed that rbIFN-tau also decreased PMA-stimulated PG production and COX-2 protein. It can be concluded that rbIFN-tau inhibition of OT-stimulated PG production is due to down-regulation of OT receptor, COX-2, and PGFS. (+info)
Steroid regulation of retinol-binding protein in the ovine oviduct.
Two studies were conducted to identify retinol-binding protein (RBP) expression in the ovine oviduct and to determine the role of ovarian steroids in its regulation. Ewes were salpingectomized on Days 1, 5, or 10 of their respective estrous cycles, and oviducts were homogenized for RNA analysis, fixed for immunocytochemistry (ICC), or cultured for 24 h for protein analysis. ICC localized RBP to the epithelium of all oviducts. RBP synthesis was demonstrated by immunoprecipitation of radiolabeled RBP from the medium of oviductal explant cultures. Explant culture medium from oviducts harvested on Day 1 contained significantly more RBP than medium from oviducts collected on Days 5 or 10. Slot-blot analysis demonstrated that steady-state RBP mRNA levels were significantly higher on Day 1 than Day 5 or 10. In the second experiment, ovariectomized ewes were treated with estradiol-17beta (E2), progesterone (P4), E2+P4 (E2+P4), or vehicle control, and oviducts were analyzed as above. P4 alone or in combination with E2 significantly reduced steady-state RBP mRNA levels compared to those in E2-treated animals. Oviductal explants from E2- and E2+P4-treated animals released 3- to 5-fold more RBP into the medium than control and P4 treatments as determined by ELISA. RBP synthesis of metabolically labeled RBP was increased by E2 and E2+P4 treatments. This study demonstrates that P4 applied on an estradiol background negatively regulates RBP gene expression in the oviduct whereas estradiol appears to stimulate RBP synthesis and secretion. (+info)
Luteal regression in the normally cycling rat: apoptosis, monocyte chemoattractant protein-1, and inflammatory cell involvement.
In hypophysectomized rats, prolactin induces regression of the corpora lutea. Luteal regression is accompanied by infiltration of monocytes/macrophages, declines in luteal mass and plasma progestins, and increased staining for monocyte chemoattractant protein-1 (MCP-1). We investigated whether similar events are induced during the estrous cycle, after the proestrous prolactin surge. Rats were killed on proestrus or on estrus, and one ovary was frozen for immunohistochemical detection of MCP-1, monocytes/macrophages (ED1-positive), and differentiated macrophages (ED2-positive) and for in situ detection of apoptotic nuclei. Corpora lutea of the current (proestrus) or preceding (estrus) cycle were dissected from the ovaries of additional rats and frozen for the same analyses and for determination of total protein content. In sections of whole ovaries, intensity and distribution of MCP-1 staining were increased in corpora lutea of multiple ages on estrus as compared to proestrus, as were numbers of differentiated macrophages and apoptotic nuclei per high-power field. Sections of isolated corpora lutea showed these increases on estrus, and the number of monocytes/macrophages per high-power field was also significantly increased. Accompanying these inflammatory/immune events, the corpora lutea on estrus showed decreased weight and total protein per corpus luteum, as compared to corpora lutea on proestrus. These changes are consistent with a proposed role for prolactin in the initiation of luteal apoptosis and of a sequence of inflammatory/immune events that accompany regression of the rat corpus luteum during the normal estrous cycle. (+info)
Precocious estrus and reproductive ability induced by PG 600 in prepuberal gilts.
A total of 29 SPF Large White prepuberal gilts (mean age 152 days at treatment) were examined for estrous and ovulatory responses after PG 600 treatment. After treatment, 85.2% of the gilts showed standing estrus within 6 days. Whereas the treatment-to-estrus interval and duration were 3.7 and 1.9 days respectively. As ovulation occurred on Day 5 to 6, appropriate timing of artificial insemination would be about 4 days after treatment. Fertility of gilts revealed to be excellent, giving rise to a high percentage of normal embryos, 85.3%. Meanwhile, development and growth of fetuses were mostly normal. Other reproductive performances recorded were: mean litter size 6.8; mean birth weight 1.26 kg; weaning-to-return estrus interval 5 to 8 days. In conclusion, PG 600 was found to be useful in inducing fertile estrus in prepuberal gilts, a result which will be of interest for commercial pig farms. (+info)
Expression of the oxytocin receptor in relation to steroid receptors in the uterus of a primate model, the marmoset monkey.
The dynamics of the receptors for oestrogen (ER), progesterone (PR) and oxytocin (OTR) in the marmoset uterus have been analysed throughout the entire cycle and early pregnancy. Uteri obtained during the early, mid/late and late proliferative phase, and the early, mid and late secretory phase and early pregnancy were examined by immunohistochemistry (OTR, ER, PR) and autoradiography (OTR). A massive upregulation of the ER in the cell nuclei of glandular epithelium and stromal cells during the mid proliferative phase was succeeded by a declining staining intensity and positively stained cell number in the secretory phase. PR immunoreactivity increased in the late proliferative phase and early secretory phase, mainly within the cell nuclei, and then declined in both intensity and cell number towards the mid to late secretory phase. Myometrium showed a similar staining pattern for the steroid receptors. OTR were expressed weakly in stroma throughout the entire cycle, increasing slightly in the secretory phase. Glandular epithelium showed positive staining only during the periovulatory period. Myometrial OTR expression was weak during the proliferative phase, increased towards the secretory phase, and was maximal in the late secretory phase. Myometrial tissue adjacent to endometrium was most strongly stained. A cyclic shift evidently occurred in the pattern of steroid receptors, perhaps reflecting the steroid environment or the luteinizing hormone increase associated with ovulation. (+info)
Gonadotropin-releasing hormone improves reproductive performance of dairy cows with slow involution of the reproductive tract.
Eighty multiparous Holstein cows were assigned randomly at calving to receive either 100 microg of GnRH or saline 13 or 14 d postpartum (PP). From 4 to 28 d PP the cows' reproductive organs were palpated weekly per rectum, and cows were subclassified within each group as undergoing slow (delayed) cervical and uterine involution (abnormal) or as normal cows. Last milk obtained after removing the milking machine was assayed for progesterone 3 times a week for 120 d PP. Fourteen of the 80 cows were removed from the experiment because of culling or various veterinary treatments of pathologic conditions that could confound analysis of the GnRH treatment effects. As expected, the treatment of normal cows with GnRH had no significant effects on the first estrus or the first estrous cycle PP, on services per conception, days open, or any other reproductive trait measured. However, in the abnormal group of cows receiving saline, first rebreeding after calving was delayed (81 vs. 67 d), fewer were pregnant by 105 d PP (23 vs. 64%), and number of days open was greater (121 vs. 87 d) compared with those receiving GnRH; all were significant (P<.05). Treated abnormal cows were equivalent to the control normal cows. Thus, GnRH given 13 to 14 d PP to cows characterized as undergoing slow involution of the reproductive system, but with no other clinical problems, seems to assist in promoting rapid normal reproductive function. Subsequent losses due to culling were greatly reduced. (+info)
Effects of twinning on postpartum reproductive performance in cattle selected for twin births.
The effects of twinning, dystocia, retained placenta, and body weight on postpartum reproduction were evaluated for 3,370 single and 1,014 twin births. Females were bred by AI for 40 d followed by 20 or 30 d of natural service with equal numbers bred and calved in spring and fall. Percentage of dams cyclic by the end of the AI period was lower (P<.05) for dams birthing and nursing a single calf (92.4%) than for dams birthing twins and nursing zero (98.7%) or two (94.7%) calves. Whereas the interval from parturition to first estrus was shorter (P<.01) for dams birthing and nursing a single (56.9 d) than for dams birthing twins and nursing one (68.5 d) or two (69.6 d) calves, length of the interval was further reduced by dystocia in nonlactating dams of either twins or singles (type of birth x dystocia, P<.05). Ensuing pregnancy rates were also affected by type of birth and dystocia. Without dystocia, dams birthing and nursing a single calf had a higher pregnancy rate (79.2%) than dams birthing twins and nursing one (61.7%) or two (66.3%) calves, whereas the lower ensuing pregnancy rates associated with dystocia in dams of singles (71.9%) resulted in similar rates among dams of singles and twins with dystocia (type of birth x dystocia; P<.01). Having a retained placenta resulted in a lower incidence of (93.5 vs. 96.4%, with vs. without; P<.05) and a longer interval to (64.7 vs. 59.2 d; P<.01) estrus while reducing subsequent pregnancy rates (X = 9.6%) in 3 of the 7 yr evaluated (retained placenta x year, P<.01). Because all parous females were bred during the same calendrical period, the shorter gestation length for twin calves (275.6 vs. 281.3 d) resulted in a longer interval from parturition to conception for twin births, whereas means for conception date differed by only 2 d between dams of twins and singles. Furthermore, a reduction (P<.01) in the interval to conception occurred with dystocia in dams of singles (89.3 vs. 85.0 d, without vs. with dystocia) and of twins nursed by zero (116.9 vs. 83.5 d), one (100.2 vs. 92.8 d), or two (96.1 vs. 97.2 d) calves. Another detriment to fertility was the higher incidence of fetal mortality or abortions associated with twin vs. single pregnancies (12.4 vs. 3.5%; P<.01). However, despite the lower conception rates for dams of twins, the increased prolificacy provides an opportunity to increase total beef production with a twinning technology. (+info)