Endometrial decidualization does not trigger the blood pressure decline of normal early pregnancy in mice

A drop in mean arterial pressure (MAP) characterizes early, normal pregnancies of humans and of inbred mice, species with hemochorial placentation. Murine MAP, assessed by continuous radiotelemetry, falls from implantation to Gestation Day 9 (GD9) and then recovers. The change in the trajectory of mouse MAP after GD9 coincides with full maturity of the placenta and onset of its circulation. To identify whether these early gestational changes in hemodynamic function are conceptus and/or maternally regulated, pseudopregnancy (conceptus absent) with endometrial decidualization was established in radio transmitter-implanted, randomly bred CD1 mice. To avoid destabilization of MAP by anesthesia and surgery, decidualization was induced by transcervical infusion of concanavalin A-coated Sepharose beads 48 h after the female had copulated with a vasectomized male. In comparison to the postimplantation drop in MAP recorded in CD1 females mated by fertile males, pseudopregnancy MAP was stable to Gestation-Equivalent Day 10 in mice with confirmed endometrial decidualization at euthanasia. Thus, decidualization, with its accompanying pregnancy-like endocrine environment and uterine neoangiogensis and immune cell recruitment, is inadequate to depress early postimplantation MAP. These data suggest that the physiological modulation of early gestational MAP is not driven by maternal changes but is altered through conceptus-based mechanisms.     

The effect of LH-RH administration on LH release in the female rabbit.

Intracarotid infusion of LH-RH to female rabbits stimulated a significant increase in plasma LH concentration in the jugular vein. This response varied with the reproductive state of the animal, with a greater release occurring in oestrous (spontaneous or oestrogen-induced) and non-receptive does than in pseudopregnant or ovariectomized animals. If ovariectomized rabbits were pretreated with oestrogen, the pituitary response to LH-RH was restored. These findings suggest that there is little change in pituitary sensitivity to LH-RH infusion between oestrous and non-receptive rabbits, although pseudopregnancy (high physiological levels of progesterone) or ovariectomy inhibit its ability to respond to a releasing-hormone stimulus.     

The Acute Phase Protein Ceruloplasmin as a Non-Invasive Marker of Pseudopregnancy, Pregnancy, and Pregnancy Loss in the Giant Panda

After ovulation, non-pregnant female giant pandas experience pseudopregnancy. During pseudopregnancy, non-pregnant females exhibit physiological and behavioral changes similar to pregnancy. Monitoring hormonal patterns that are usually different in pregnant mammals are not effective at determining pregnancy status in many animals that undergo pseudopregnancy, including the giant panda. Therefore, a physiological test to distinguish between pregnancy and pseudopregnancy in pandas has eluded scientists for decades. We examined other potential markers of pregnancy and found that activity of the acute phase protein ceruloplasmin increases in urine of giant pandas in response to pregnancy. Results indicate that in term pregnancies, levels of active urinary ceruloplasmin were elevated the first week of pregnancy and remain elevated until 20–24 days prior to parturition, while no increase was observed during the luteal phase in known pseudopregnancies. Active ceruloplasmin also increased during ultrasound-confirmed lost pregnancies; however, the pattern was different compared to term pregnancies, particularly during the late luteal phase. In four out of the five additional reproductive cycles included in the current study where females were bred but no birth occurred, active ceruloplasmin in urine increased during the luteal phase. Similar to the known lost pregnancies, the temporal pattern of change in urinary ceruloplasmin during the luteal phase deviated from the term pregnancies suggesting that these cycles may have also been lost pregnancies. Among giant pandas in captivity, it has been presumed that there is a high rate of pregnancy loss and our results are the first to provide evidence supporting this notion.     

Transformation of endometrium and fertility in late stages of pseudopregnancy in the rabbit

Ovulation was induced in rabbits between Days 14 and 18 of pseudopregnancy by an intravenous injection of hCG. Induction of ovulation from Day 16 onwards led to normal progestational endometrial transformation. In rabbits injected on Day 14 or 15, a normal preimplantation endometrial morphology developed, but not earlier than 7 days after hCG (Day 14/15 + 7). Uteroglobin secretion was advanced during the second pseudopregnancy. After mating or artificial insemination, fertility was greatest on Day 18 of pseudopregnancy. Conception failed on Day 14 and embryo transfers were unsuccessful on Day 14 + 1. Transfers performed on Day 14 + 3, however, led to implantation and offspring, even though endometrial morphology did not correspond to the normal Day 3 preimplantational morphology at the time of transfer. We conclude that endometrial transformation typical of normal pseudopregnancy can be induced by ovulation during the regeneration phase of pseudopregnancy from Day 16 onwards; fertilization and implantation can be achieved as early as Day 15 of pseudopregnancy; an oestrous period with high mating activity and fertility occurs about 3 days later; and Day 14 after hCG represents a limited time of functional change from pseudopregnancy to a fertile uterine cycle in the rabbit.     

1,25-Dihydroxyvitamin D3 induces in vivo the decidualization of rat endometrial cells

Intraluminal injection of female rats at Day 5 of pseudopregnancy with 10-500 ng 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) significantly increased the uterine weight and induced decidual reaction. This effect was observed as early as the 3rd day after 1,25-(OH)2D3 injection. It was detectable only in the injected left horn and not in the non-injected right horn. A 500 ng dose of 25-(OH)D3 had no such effect. The present in-vivo results suggest that 1,25-(OH)2D3 may play a physiological role in endometrial cell differentiation into decidual cells, a crucial step in the process of blastocyst implantation.     

Uterine adenylate cyclase activity during the estrous cycle and early progestation in the rat: responses to fluoride activation and decidual induction

Uterine adenylate cyclase (AC) activity of the rat was measured by radiochemical analysis during the estrous cycle and early pseudopregnancy. During the estrous cycle, AC activity increased from 4.6 to 16.9 pmol cAMP formed/min X mg protein between metestrus and proestrus. Although AC was activated 2- to 3-fold at all cycle stages by 10 mM NaF, the resulting pattern of activity was similar to that measured in the absence of fluoride. The results demonstrated that the pattern of AC activity during the cycle was similar to that of other estrogen-sensitive uterine enzymes and that the ovarian hormones probably altered enzyme biosynthesis and turnover to a greater extent than activation and kinetic properties. Following the induction of pseudopregnancy by cervical stimulation, enzymic activity increased from 3.5 to 9.4 pmol between Days 1-4 (Day 1=leukocytic vaginal smear) and declined thereafter. AC activity was increased 2- to 5-fold by NaF on all days. AC activity was similarly increased by a mechanical trauma to the uterus, but only when the trauma was applied on Day 4. Following trauma to the uterus, AC activity was not increased further by NaF. The similarities between the physicochemical characteristics of AC during the estrous cycle and early progestation suggested that the enzyme during all endocrine states had virtually identical properties. However, the transient sensitivity to activation after trauma on Day 4 was unique to progestational uteri. Because the properties of enzyme were not altered by the endocrine state of the tissue, the transient sensitivity to activation by trauma was suggested to be a result of hormone-induced alterations in the membrane in which AC is sequestered.     

Effects of (-)-trans- delta 9-tetrahydrocannabinol on serum prolactin in the pseudopregnant rat

Groups of pseudopregnant rats were injected intravenously with (-)-trans- delta 9-tetrahydrocannabinol (THC) to determine its effects on serum prolactin (PRL) and the maintenance of pseudopregnancy. A single injection of 4 mg THC/kg BW at 2400 h on the first day of leukocytic vaginal smears of pseudopregnancy (D-1) delayed the ensuing nocturnal PRL surge for approximately one hour. When THC (1.0 mg/kg BW) was administered hourly from 2400 h on D-1 through 1700 h on D-2, the nocturnal surge was blocked and serum PRL levels were suppressed until 0600 h on D-2, but not thereafter. Neither treatment altered the duration of pseudopregnancy. These results indicate that the nocturnal surge secretion of PRL during early pseudopregnancy in the rat is sensitive to THC suppression, but that this suppression is not adequate to influence the duration of pseudopregnancy. The mechanism through which THC exerts this action remains unknown.     

Regulation of adenylate cyclase activity and stimulation response in relation to endometrial receptivity in the rabbit

Adenylate cyclase activity was measured in broken cell preparations of whole endometrial tissue from rabbits on Days 0, 1, 6.5, 9 and 15 of pseudopregnancy and in endometrial epithelial and stromal cells on Days 1 and 6.5 to assess the specific response of individual cell types. In dispersed cells, adenylate cyclase activity was higher (P less than 0.01) in stromal than in epithelial cells and reduced on Day 6.5 compared to Day 1 in both cell types. The response of adenylate cyclase to isoproterenol appeared more important relative to the PGE-2 response in epithelial than in stromal cells and strongly reduced in the former on Day 6.5. In endometrium, the overall adenylate cyclase activity was increased significantly on Day 1 of pseudopregnancy compared to Day 0 (oestrus), only 18 h after injection of hCG. On the following days, the activity decreased progressively on Days 6.5 and 9 and exhibited a recovery on Day 15. Adenylate cyclase response to isoproterenol (% over GTP) was comparable on Days 0, 1 and 6.5, abolished on Day 9 and recovered on Day 15. Maximal response to PGE-2 (% over GTP) was observed on Day 6.5, at the time of implantation, maintained on Day 9 and reduced on Day 15 towards the low levels measured in oestrus and Day 1 of pseudopregnancy. Our results demonstrate a dramatic alteration of adenylate cyclase activity in rabbit endometrium during pseudopregnancy. It suggests a possible involvement of catecholamines and prostaglandin E-2 in the regulation of endometrial receptivity through a cAMP-mediated process.     

Participation of ovarian 20 alpha-hydroxysteroid dehydrogenase in luteotrophic and luteolytic processes during rat pseudopregnancy

In normal rats, before Day 12 of pseudopregnancy, minimal levels of 20 alpha-HSD activity were detected in functional CL whereas those in the residue were 3-5 times higher. When ovulation was blocked for more than 2 weeks by placing rats in a continuously lit environment before the induction of pseudopregnancy, only minimal levels of 20 alpha-HSD activity were detectable in the functional CL and residue before Day 12. In normal pseudopregnant rats, there was a linear increase in 20 alpha-HSD activity from Day 12 to 15 in the functional CL and residue, but the rate of elevation was much higher in functional CL. This tendency was much more clear-cut in rats in the continuous lighting. In immature rats in which pseudopregnancy was induced by PMSG and hCG treatment, 20 alpha-HSD activity peaked twice. The first small peak was attributed to the early regression of some of the large number of corpora lutea, and the changes in 20 alpha-HSD activity in most of the corpora lutea paralleled those in rats in continuous lighting. Bromocriptine abolished the prolactin surges, and in normal pseudopregnant rats an increase in 20 alpha-HSD activity in functional CL started from 12 h and the rate of the increase was accelerated from 36 h afterwards, while a relatively small increase was observed in the residue at 18 h and later.(ABSTRACT TRUNCATED AT 250 WORDS)     

蜱雄性附腺分泌物及其功能的研究进展

交配是蜱类繁殖过程的关键环节,诱发雌蜱发生一系列的生理变化,并最终产卵。蜱雄性附腺分泌物在交配过程中发挥着重要作用,具有保护、活化精子,促进受精、卵巢发育和卵黄发生的功能,并对雌蜱的生殖生理行为等产生影响,如诱导雌蜱快速吸血和加速产卵。本文在简要分析蜱雄性附腺结构和分泌物生化特性基础上,系统阐述了蜱雄性附腺分泌物中各种功能因子的研究现状,着重论述其在精子获能、诱导雌蜱吸血、促进雌蜱卵巢发育和卵黄发生等方面的进展,并对未来研究提出了展望,以期为此领域的研究拓展思路。     

Blockage of induced pseudopregnancy by electrochemical stimulation of the limbic system.

Pseudopregnancy induced by cervical stimulation was inhibited by acute electrochemical stimulation of the corticomedial amygdala or dorsal hippocampus under sodium pentobarbital anesthesia (40 mg/kg) in adult, cyclic female Sprague-Dawley rats. The degree to which pseudopregnancy was blocked depended on temporal conditions of brain stimulation and sodium pentobarbital administration. Pentobarbital alone had a suppressing effect on the incidence of pseudopregnancy, especially when it preceded cervical stimulation. Limbic stimulation before cervical stimulation had a tendency to potentiate the suppression of pseudopregnancy by pentobarbital. After cervical stimulation, hippocampal stimulation tended to inhibit the development of pseudopregnancy, potentiating the pentobarbital suppression, while amygdala stimulation tended to override the pentobarbital blockage of pseudopregnancy. These findings suggest a negative influence of these two limbic structures and pentobarbital on the secretion of prolactin.     

Nocturnal surges and reflexive release of prolactin in parentally behaving virgin female and male rats

Maternally behaving virgin rats are capable of releasing prolactin reflexively in response to stimulation by pups, especially during the proestrous/estrous phase of the cycle. When such rats are chronically exposed to pups they usually undergo a state of pseudopregnancy during which prolactin is secreted in a pattern of nocturnal surges. The present series of experiments evaluated the initiation of nocturnal prolactin surges in maternally behaving virgins, the role of estrogen in the reflexive release of prolactin, and the influence of gender on these two modes of prolactin secretion. It was found that the nocturnal surges of prolactin are already present on Days 1 and 2 of pup-induced pseudopregnancy. At this stage, however, the surges are not yet autonomous, seeing that pseudopregnancy is interrupted shortly after removal of the pups on Day 2. Activation by vaginocervical stimulation of the "mnemonic" neurogenic system that controls the autonomous nocturnal prolactin surges did not interfere with the reflexive pup-induced release of prolactin in maternally behaving virgins. The capacity of reflexive prolactin release in the virgin rat was abolished by ovariectomy, restored by estrogen replacement, and persisted for only 24 hr following estrogen removal. Paternally behaving males subjected to chronic exposure to pups were incapable of secreting nocturnal surges of prolactin characteristic of the pseudopregnant female. Such males were also incapable of releasing prolactin reflexively in response to stimulation by pups, even when supplemented with exogenous estrogen. These results indicate that the two modes of prolactin secretion are sex dependent, and that the maternally behaving virgin, unlike the postpartum rat, requires concurrent estrogenic facilitation for releasing prolactin in response to stimulation by young.     

Influence of time of mating and paced copulation on induction of pseudopregnancy in cyclic female rats

The present experiment was designed to examine whether changes occur during the course of behavioural oestrus in the sensitivity of the female to copulatory stimulation and in patterns of sexual behaviour which might influence the likelihood of luteal maintenance. Cyclic female rats were mated on the evening of pro-oestrus (21:00 h) or early on the morning of oestrus (05:00 h) and received either 5 or 10 intromissions from males under conditions which allowed or did not allow pacing of contacts with males to occur. During mating, the levels of sexual receptivity, the timing of sexual mounts from males, and pacing behaviours were measured. The occurrence of pseudopregnancy or pregnancy in each animal was determined by examining vaginal smears for 14 days after mating and by examining the uterus for the presence of fetuses at the end of the experiment. At both mating times, pacing of copulation with males increased the likelihood of prolonged luteal activity. However, females were more likely to become pseudopregnant following non-paced mating at 05:00 h than at 21:00 h the previous evening. Of those females receiving an ejaculation during mating, no difference were seen between groups in the incidence of pregnancy. This change in sensitivity to cervical-vaginal stimulation during oestrus was not associated with changes in sexual receptivity or pacing behaviour. The ability of cervical-vaginal stimulation to induce pseudopregnancy therefore increases toward the end of the period of oestrus, but the behavioural mechanisms which regulate receipt of such stimulation remain constant during that time.     

Apical plasma membrane-bound enzymes of rabbit uterine epithelium. Pattern changes during the periimplantation phase

In order to monitor changes in the apical cell membrane of rabbit uterine epithelium which are postulated to be a precondition for trophoblast attachment, the marker enzymes: alkaline phosphatase, aminopeptidase M, gamma-glutamyl transferase and dipeptidyl peptidase IV were investigated during the periimplantation phase. Endometrium of early pregnancy (implantation chamber, interblastocyst endometrium; 5-8 days post coitum, d p.c.) was compared with specimens obtained at hCG-induced pseudopregnancy (p. hCG) to distinguish between membrane changes regulated by maternal plasma steroid hormones and such which might be induced locally by blastocyst-derived signals. All enzymes tested showed their main activity at 5 d p.c./p. hCG. The weakest reaction in this series of stages was generally found at 8 d p.c. (interblastocyst segments) or at 8 d p. hCG. In contrast to the rest of the epithelium, the implantation chamber retained high activity of dipeptidyl peptidase IV, and the activity of alkaline phosphatase even raised here again at 7 and 8 d p.c. indicating a direct local influence of the blastocyst on the luminal epithelium. The results suggest that 1) considerable changes occur in the composition of the apical plasma membrane of the uterine epithelium when the endometrium enters the "receptive state", 2) the overall trend is towards a loss of apical-type characteristics of this membrane domain and 3) the changes are modulated both systemically (by plasma steroid hormone levels) and locally by signals from the implanting blastocyst.     

Transient DNMT1 suppression reveals hidden heritable marks in the genome

Genome-wide demethylation and remethylation of DNA during early embryogenesis is essential for development. Imprinted germline differentially methylated domains (gDMDs) established by sex-specific methylation in either male or female germ cells, must escape these dynamic changes and sustain precise inheritance of both methylated and unmethylated parental alleles. To identify other, gDMD-like sequences with the same epigenetic inheritance properties, we used a modified embryonic stem (ES) cell line that emulates the early embryonic demethylation and remethylation waves. Transient DNMT1 suppression revealed gDMD-like sequences requiring continuous DNMT1 activity to sustain a highly methylated state. Remethylation of these sequences was also compromised in vivo in a mouse model of transient DNMT1 loss in the preimplantation embryo. These novel regions, possessing heritable epigenetic features similar to imprinted-gDMDs are required for normal physiological and developmental processes and when disrupted are associated with disorders such as cancer and autism spectrum disorders. This study presents new perspectives on DNA methylation heritability during early embryo development that extend beyond conventional imprinted-gDMDs.     

Alterations in intercellular junctions of the uterine epithelium during the preimplantation phase in the rabbit

Summary In the rabbit, the pseudopregnant uterus has been used as a model for studying alterations characteristic of the preimplantation phase. Alterations in intercellular junctions of the uterine epithelium were investigated during early pseudopregnancy (day 0 to day 6) by means of the freeze-fracture technique.In the uterine epithelium of oestrous females the zonula occludens belongs to the tight type of tight junctions. During pseudopregnancy an impressive proliferation of tight junctional belts can be observed. The basal strands proliferate, forming loops perpendicular to the luminal surface, whereas the more or less parallel arrangement of the luminal strands is maintained. At day 4 of pseudopregnancy macular tight junctions begin to develop on the lower portions of the lateral plasmalemma and are extensive by day 6 post hCG.Small gap junctions are infrequent between cells of the uterine epithelium and show no significant changes during the preimplantation phase.The physiological significance of the present morphological observations is discussed in the light of changes occurring during the preimplantation period.Supported by grant Kü 210/9 from the Deutsche Forschungsgemeinschaft     

Induction of pseudopregnancy in the American Black Bear (Ursus americanus)

The aim of this study was to determine whether American Black Bears (Ursus americanus) can experience a pseudopregnancy of the same duration as pregnancy. To do this, we treated three nonmated, captive female bears with human chorionic gonadotropin (hCG) during one breeding season, and saline during another. Progesterone concentrations were measured in monthly blood samples to determine whether pseudopregnancy had occurred. Elevated progesterone concentrations were observed in two out of three bears treated with hCG. We conclude that 1) Elevated progesterone concentrations can be induced in black bears by injection of 35 U/kg hCG during the mating season. 2) Bears can experience a pseudopregnancy, identical in length to pregnancy, in which progesterone profiles are indistinguishable from those of pregnancy.     

Use of pyridoxine hydrochloride in the interruption of lactation in female dogs with pseudopregnancy

The aim of this research was to evaluate the use of pyridoxine hydrochloride and its associated side effects in the treatment of pseudopregnancy in female dogs. A total of 40 female dogs, with no defined breed, in non-gestational diestrus, with clinical complaint of milk production were selected. The female dogs were divided into four experimental groups of 10 animals each, treated orally for 20 days with 10mg/kg/day (G1) and 50mg/kg/day (G2) of pyridoxine hydrochloride (vitamin B6), 5μg/kg/day of cabergoline (G3), and with a placebo, in the case of the control group (G4). The effects of the treatments on milk production were investigated, as well as possible systemic side effects, macroscopic uterine and ovarian alterations, and uterine histology. During the investigated period, G2 and G3 were equally efficient (P>0.05) in lactation suppression, differing (P>0.05) from the other groups. There were no systemic side effects or uterine changes associated with administration of the studied drug. Vitamin B6 (50mg/kg) has shown to be a safe and economically viable alternative for lactation interruption in female dogs with pseudopregnancy.     

Changes and localization of ovarian carbonyl reductase during pseudopregnancy and pregnancy in rats

The present study investigates changes in the activity and enzyme content of ovarian carbonyl reductase (CR), which catalyzes the reduction of 9-keto and 15-ketoprostaglandins in rats during pseudopregnancy and pregnancy. The activity of ovarian CR decreased from the onset of pseudopregnancy and pregnancy, reaching 20-30% of the Day 1 value by Day 12 of pseudopregnancy and 50-60% of the Day 1 value by Day 14 of pregnancy. In the case of pregnant rats, the enzyme activity maintained a minimal level between Day 14 of pregnancy and Day 22 of parturition. An acute increase of the enzyme activity was found on the morning after parturition. The CR content in the ovary maintained a constant level from Day 1 to Day 12 of pseudopregnancy and to Day 18 of pregnancy. In pregnant rats, there was a gradual decrease after 18 days and then a surge during parturition. CR was primarily localized in interstitial gland cells and in theca interna cells but was not found in corpora lutea cells in the ovary during the estrous cycle. Additional immunostaining was also observed in corpora lutea cells during pseudopregnancy and pregnancy. The changes in ovarian CR activity, i.e. the rapid decrease with progressing pseudopregnancy and pregnancy, correlated with the increase in progesterone and the decrease in LH. These results indicate that rat ovarian CR may be regulated via the hypothalamo-pituitary-ovarian axis and may also be involved in luteal function.     

Induction of pseudopregnancy in the mongolian gerbil (Meriones unguiculatus) by vaginal stimulation.

In rats, pseudopregnancy has been induced by mating with vasectomized males, by mechanical stimulation of the uterine cervix with a glass rod or vibrator, and by stimulation of the vagina with a tampon. On the other hand, no practical data are available in reports on the induction of pseudopregnancy in Mongolian gerbils. Pseudopregnancy of gerbils has been induced by mating with vasectomized males. But this method was uncertain because the incidence of pseudopregnancy was lower than that obtained in rats by other means. In the present study, two experiments were undertaken as follows. 1) Copulatory behavior of gerbils was observed for one hour to determine the most effective stimulation interval. 2) From the results of Experiment 1, female gerbils in estrus were mechanically stimulated to test the effectiveness of inducing pseudopregnancy by vaginal stimulation at various time intervals. The results of these experiments indicated that, although the frequency of copulatory behavior varied among individuals, on average the most effective method for inducing pseudopregnancy was stimulation of 5 min duration and at 20 or 30 min intervals. Because the incidence of pseudopregnancy induced by such mechanical stimulation (83.3%) was higher than that induced by mating with vasectomized males (30.0%), this method might be useful in inducing pseudopregnancy in Mongolian gerbils.