Pregnancy blockage and the memory of the stud male in the vole (Microtus agrestis).

When female voles were allowed contact with the stud male for only 1 h at the time of mating, 55% exhibited pregnancy failure when exposed to a strange male 48 h later. When females were made psuedopregnant by hormone treatment and vaginal stimulation (i.e. no stud male involved), 87% exhibited luteal failure when exposed to a strange male. It is suggested that the characteristics of the stud male are rapidly imprinted upon the female at the time of mating and that this imprinting is important in preventing the female showing a blocking response to this male upon any subsequent exposure.     

Influence of pre- and post-copulatory pair contact on pregnancy success in Djungarian hamsters, Phodopus campbelli

Females housed with their mates for 3 or 4 days before mating took place (i.e. early in the oestrous cycle at the time of introduction to the mate) were significantly more likely to litter than were females housed with their mates for only 1 or 2 days before mating. The duration of post-copulatory pair contact had a complex effect on pregnancy success. While only 41% of females littered when they had 24 h of post-copulatory pair contact, females exposed to either longer or shorter durations of post-copulatory pair contact littered at significantly higher rates. Exposure to a strange male 24-48 h after mating did not produce a strange-male induced pregnancy block. The critical parameter responsible for the decrease in the number of females littering was the absence of the mate, irrespective of the presence or absence of a strange male. If this pattern of pregnancy block is adaptive for females, it seems probable that females in the wild require substantial levels of paternal investment by their mates.     

Stud male-induced protection of implantation in food-deprived mice: masking effect of an artificial scent on pheromonal odour

In contrast to unscented stud males, stud males anointed with a commercial perfume failed to protect implantation in food-deprived females. It is suggested that the failure of perfumed stud males to protect pregnancy in their coital partners is due to the masking effect of the perfume on the stud male-originating olfactory cue which stimulates luteotrophic activity in females. The results are also consistent with the view that the newly inseminated female mouse identifies the stud male as an individual through a pheromonal cue and this is involved in the protective effect on implantation in the food-deprived female.     

Induction of oestrus in the camel (Camelus dromedarius) during seasonal anoestrus

Injection of 7000 i.u. PMSG induced oestrus in 7 camels during the last part of seasonal anoestrus. Mature follicles developed and a CL was formed after fertile mating. However, pregnancy was not maintained by Day 60 in the 3 females detected as pregnant by rectal palpation and increased progesterone concentrations at Day 50. A single male camel mated with 4 of the females 2-16 days after the PMSG injection, and 2 or 3 matings occurred. The failure of pregnancy after induction of oestrus and mating during seasonal anoestrus was probably due to inadequate luteal function.     

Luteotrophic influence of early bovine embryos and the relationship between plasma progesterone concentrations and embryo survival

This study was carried out to evaluate the luteotrophic influence of early (before Day 7 as well as after Day 7; Day 0=estrus) bovine embryos and the relationship between plasma progesterone (P4) concentrations and embryo survival. Virgin Holstein dairy heifers (n=325) from a single herd were randomly allocated to be nonbred, bred by artificial insemination (AI) or by embryo transfer (ET). Bred heifers were either treated with 1500 IU human chorionic gonadotrophin (hCG) on Day 7 of the estrous cycle or received no hCG treatment. Plasma P4 concentrations on Days 0, 5, 7, 10, 13, 15, 17, 19 and 21 were similar in pregnant AI- and ET-bred heifers and, this was observed in both hCG-treated and untreated females. Nonbred, AI- and ET-bred nonpregnant heifers (both hCG-treated and untreated) presented similar plasma P4 concentrations. Plasma P4 concentrations of pregnant heifers significantly deviated from those of nonpregnant and nonbred heifers on Day 17. In hCG-treated heifers, plasma P4 concentrations and Day 28 pregnancy rate were significantly higher in females with an induced accessory corpus luteum (CL) than in those females without an induced accessory CL. Treatment with hCG, although inducing the formation of accessory CL and significantly increasing plasma P4 concentrations had no significant effect on Day 28 pregnancy rate. In conclusion, this study does not support the existence of any peripherally detectable luteotrophic influence from early embryos (Days 5-7). Plasma P4 was only significantly related to embryo survival on Day 17, the time of expected onset of luteolysis.     

Studies of pericopulatory pregnancy blockage and the gestation period in deer mice (Peromyscus maniculatus)

It has previously been shown that female deer mice in cycling estrus that copulate with or are exposed to more than one male at the time of mating have a lower probability of delivering a litter than when mating with and being exposed to just one male. In the present research the effect of post-mating exposure to the bedding of a strange versus a familiar male was studied in females in postpartum estrus. In four experiments females in postpartum estrus were protected from pericopulatory pregnancy blockage, presumably by suckling-induced elevated prolactin levels. Varying female experience, number of ejaculations, and the presence of suckling pups had no effect on this protection. Thus, both the pericopulatory block and the Bruce effect (a more delayed type of pregnancy blockage) occur in cycling, but not suckling, females. This suggests that the two may have a common mechanism and leads to a new perspective in the search for adaptive significance. In addition, the subsequent gestation periods were prolonged in females nursing young; the length was affected by the number of pups weaned from the previous litter and the number of pups being carried.     

Relative roles of oestradiol and of the uterus in the maintenance of the corpus luteum in the pseudopregnant brown hare (Lepus europaeus)

Brown hares were made pseudopregnant by sterile matings or PMSG-hCG treatment (day of mating or hCG injection = Day 0 of pseudopregnancy). Progesterone secretion by the CL began 3-4 days after the ovulatory stimuli, reached maximum on Days 8 to 11 and decreased thereafter to reach low levels from Day 9 to 18, depending on the female. Cauterization of all large ovarian follicles on Day 7 resulted in an immediate luteolysis in young females, but had no effect in older ones. Oestradiol capsules implanted from Day 7 to Day 46 were able to maintain progesterone secretion until at least Day 30, in intact females as well as in females with all large follicles cauterized. Hysterectomy on Day 7 or 8 was followed by an immediate drop in progesterone concentrations; oestradiol capsules implanted at the time of hysterectomy prevented the drop in progesterone values, which remained elevated until Day 38. The induction of ovulation in females hysterectomized 2 months before resulted in CL of slightly shortened life-span. The injection of PGF-2 alpha on Day 7 of pseudopregnancy was followed by an immediate luteolysis. These results suggest that oestradiol secreted by the large ovarian follicles is the main luteotrophic factor in the brown hare. In old hares, the large amount of interstitial tissue could secrete oestrogens, and thus maintain pseudopregnancy. On Day 7 of pseudopregnancy, the uterus secretes a luteotrophic substance acting either directly on the ovary, or via the pituitary, to maintain oestradiol secretion by the follicles. In long-term hysterectomized females, the CL would be able to develop independently of any trophic substance, but for a reduced duration.     

Changes in progesterone levels in pregnancy block caused by the proximity of a strange male and by the injection of PMSG

Attempts were made to examine progesterone levels during pregnancy block, first initiated by pheromones and then by the injection of pregnant mare serum gonadotropin (PMSG). The proximity of a strange male inhibits the usual increase in progesterone levels, resulting in no activation of the corpora lutea, from the estrous cycle to the pregnancy. Strain differences were observed in the imitation of pregnancy block by the injection of PMSG. The C3H and BALB/c strains required much smaller doses of PMSG for pregnancy block than the C57BL/6 strain did. The changes of progesterone observed in pregnancy block initiated by PMSG were slightly different from that caused by the proximity of the strange male: The progesterone levels temporally increased on the day following the injection of PMSG and then decreased on day 4 in the PMSG-treated group. These results suggest that PMSG imitates the effect of the proximate strange male in the pregnancy block.     

Social interactions associated with reproduction in the bushveld gerbil <Emphasis Type="Italic">Gerbilliscus leucogaster</Emphasis>

The study of social behaviour can give us important insights into the social and mating system of a species or population. We investigated male–female interactions in captive bushveld gerbils, Gerbilliscus leucogaster, to gain insight into the social behaviour and mating structure. We conducted two experiments. In experiment 1, we studied the interactions between two strangers for a week in a three-tank setup, which tested behavioural variation spatially and temporally. Although tolerance between strange males and females increased with time, it remained generally low throughout the experiment. Females appeared to be cautious of males initially, but they later became aggressive towards them. Males showed an increase in submissive behaviour over time. The ‘home’ cage did not appear to be defended by either sex. In experiment 2, we investigated the social interactions of male–female pairs during pregnancy and lactation. Aggression persisted throughout the study and amicable behaviour was low; females were much more aggressive than males. We did not detect changes in social behaviour with the progression of pregnancy and lactation. We suggest that pair bonding is unlikely and that promiscuity is the most probable mating system. Female aggression may be related to mate choice prior to mating and mate exclusion thereafter, while it may be a response to infanticide risk during pregnancy and lactation.     

Bromocriptine-induced implantation failure in mice: failure of the stud male to protect pregnancy.

The presence of the stud male failed to prevent implantation failure in newly inseminated females induced by administration of bromocriptine. This is in contrast with the ability of the stud male to prevent implantation failure in females induced by alien male exposure, and nutritional stress. Since bromocriptine is a potent inhibitor of hypophysial prolactin release by virtue of its stimulatory effect on hypothalamic dopaminergic activity, the results suggest that the stud male-originating luteotrophic stimulus is incapable of overriding the dopaminergic activity in bromocriptine-treated females.     

Mating Increases Neuronal Tyrosine Hydroxylase Expression and Selectively Gates Transmission of Male Chemosensory Information in Female Mice

Exposure to chemosensory signals from unfamiliar males can terminate pregnancy in recently mated female mice. The number of tyrosine hydroxylase-positive neurons in the main olfactory bulb has been found to increase following mating and has been implicated in preventing male-induced pregnancy block during the post-implantation period. In contrast, pre-implantation pregnancy block is mediated by the vomeronasal system, and is thought to be prevented by selective inhibition of the mate’s pregnancy blocking chemosignals, at the level of the accessory olfactory bulb. The objectives of this study were firstly to identify the level of the vomeronasal pathway at which selective inhibition of the mate’s pregnancy blocking chemosignals occurs. Secondly, to determine whether a post-mating increase in tyrosine hydroxylase-positive neurons is observed in the vomeronasal system, which could play a role in preventing pre-implantation pregnancy block. Immunohistochemical staining revealed that mating induced an increase in tyrosine-hydroxylase positive neurons in the arcuate hypothalamus of BALB/c females, and suppressed c-Fos expression in these neurons in response to mating male chemosignals. This selective suppression of c-Fos response to mating male chemosignals was not apparent at earlier levels of the pregnancy-blocking neural pathway in the accessory olfactory bulb or corticomedial amygdala. Immunohistochemical staining revealed an increase in the number of tyrosine hydroxylase-positive neurons in the accessory olfactory bulb of BALB/c female mice following mating. However, increased dopamine-mediated inhibition in the accessory olfactory bulb is unlikely to account for the prevention of pregnancy block to the mating male, as tyrosine hydroxylase expression did not increase in females of the C57BL/6 strain, which show normal mate recognition. These findings reveal an association of mating with increased dopaminergic modulation in the pregnancy block pathway and support the hypothesis that mate recognition prevents pregnancy block by suppressing the activation of arcuate dopamine release.     

Hormonal regulation and function of an RNA helicase,Ddx5 in corpus luteum of adult Wistar rats

Corpus luteum (CL) is an endocrine tissue involved in regulation of reproductive cycle and early pregnancy establishment. In the present study DEAD-box helicase-5 (Ddx5), a member of the DEAD box family of RNA helicases was investigated for its expression, regulation and function in CL of Wistar rats. Ddx5 was expressed in adult rat CL. Primary cell culture from supra-ovulated ovaries were established for in vitro studies. Addition of luteinizing hormone (LH; 100 ng/ml), a luteotrophic factor in primary cell culture, decreased Ddx5 RNA expression (foldchange:0.6 ± 0.075) while prostaglandin alpha (PGF_2α; 1μM), a luteolytic factor caused an increase (foldchange:2.4 ± 0.4) compared to control group. Under in vivo conditions, the administration of PGF_2α or gonadotropin-releasing hormone antagonist; cetrorelix (CET) caused luteolysis as well as an increase in the protein level of Ddx5 (foldchange:1.9 ± 0.27 and 1.4 ± 0.09 viz.; p < 0.05) in CL of adult rats. LH was administered post CET treatment which suppressed Ddx5 protein expression (foldchange:0.8 ± 0.16; p < 0.05) compared to CET treated group. Further, it was observed that the expression of Ddx5 was upregulated (foldchange:1.5 ± 0.23; p < 0.05) in CL during late pregnancy compared to mid pregnancy concomitant to luteolysis in adult rats. Overall, the results suggest for the first time that Ddx5 is expressed in rat CL and regulated by luteolytic and luteotrophic factors in an inverse fashion. Further, the data significantly correlates ddx5 expression to CL regression suggesting involvement of ddx5 in luteolysis. These results suggest a significant role of Ddx5 in female reproduction biology and warrant in depth examination of the function of Ddx5 in CL.     

Trophoblastin, an antiluteolytic protein present in early pregnancy in sheep.

Trophoblastin, an antiluteolytic component from the embryo, was identified in the ewe by the means of intrauterine injections of homogenates from trophoblasts at 14--16 days pregnancy. Homogenates from embryos and their membranes at 21--23 days pregnancy did not extend the life of the corpus luteum, suggesting that trophoblastin synthesis occurs for only a short period. The trophoblastin was thermolabile (80 degrees C for 30 min) and inactivated by pronase. Treatment of ewes with oCS, hCG, and extracts of 120-day placentae did not affect the time of luteolysis. The protein appears to be insoluble at pH 7 or 8, but to dissolve readily at pH 9.6. After injection of homogenates or extracts from 15--16-day-old trophoblasts, the initial CL were maintained for more than 1 month in most cyclic recipient ewes. Surgical removal of embryos at 21--23 days resulted in luteal maintenace for more than 1 month in over 50% of the operated animals. All the maintained CL were secretory although their average weight was about one-half of that CL of normal pregnancy, suggesting the existence of complementary luteotrophic placental factors. The uteri of most of these pseudopregnant ewes were distended with a clear, sterile fluid.     

Development of partner preferences in female prairie voles (Microtus ochrogaster): the role of social and sexual experience.

Prairie voles (Microtus ochrogaster) exhibit a monogamous mating system characterized by long-term pair bonds between mates. The purpose of this study was to examine the effect of cohabitation time and sexual experience on the development of pair bond formation in female prairie voles. Females that were allowed to cohabit for 24 hr or more, with or without mating, exhibited a strong social preference for a familiar partner versus a strange male. Females that cohabited and mated for 6 hr showed strong preferences for a familiar partner, while cohabitation for less than 24 hr, without mating, did not result in preferences for the familiar male. These results indicate that mating was not essential for partner preference formation; however, preferences developed more rapidly when mating occurred.     

Investigations on the Olfactory Discrimination Exhibited by the Females in the Male-induced Implantation Failure (the Bruce Effect) in Laboratory Mice

Direct contact with the alien male pheromone for 7 days before mating significantly decreased the rate of male-induced implantation failure (the Bruce effect) in newly inseminated female mice. By contrast, females housed with alien males for 7 days before mating so that physical contact with the latter was prevented exhibited a high rate of implantation failure following exposure to alien males after mating. Exposure to an air-borne pheromone produced by alien males induced oestrus (the Whitten effect) in a significant number of grouped females. The results do not support the suggestion that the inability of the stud male to block implantation is due to his original induction of oestrus. The results also do not support the view that the key factor in the induction of the Bruce effect is the ability of the female to identify the stud as an individual which prevents her from responding to him but allows her to react to a new male with hormonal changes leading to implantation failure and return to oestrus. It is suggested that the olfactory discrimination exhibited by the female in the Bruce effect is influenced by her overall olfactory experience before and after mating. The results are consistent with the view that the male-originating pheromone involved in the Bruce effect is distinct from the one involved in the Whitten effect.     

Studies on the control of the corpus luteum in the vole, Microtus agrestis.

The lifespan of corpora lutea resulting from hormonally induced ovulations was prolonged by exogenous prolactin, concurrent lactation, or pregnancy. Treatment of mated females with bromocriptine resulted in failure of luteal function only when the drug was given before Day 6 of pregnancy. Pregnancy was dependent on the presence of the ovaries in its later stages. The results suggest that prolactin is luteotrophic in early pregnancy but that a placental luteotrophin may become effective by Day 6 of pregnancy.     

Early maternal investment in mice: no evidence for compatible-genes sexual selection despite hybrid vigor

Confronting a recently mated female with a strange male can induce a pregnancy block ('Bruce effect'). The physiology of this effect is well studied, but its functional significance is still not fully understood. The 'anticipated infanticide hypothesis' suggests that the pregnancy block serves to avoid the cost of embryogenesis and giving birth to offspring that are likely to be killed by a new territory holder. Some 'compatible-genes sexual selection hypotheses' suggest that the likelihood of a pregnancy block is also dependent on the female's perception of the stud's and the stimulus male's genetic quality. We used two inbred strains of mice (C57BL/6 and BALB/c) to test all possible combinations of female strain, stud strain, and stimulus strain under experimental conditions (N(total) = 241 mated females). As predicted from previous studies, we found increased rates of pregnancy blocks if stud and stimulus strains differed, and we found evidence for hybrid vigour in offspring of between-strain mating. Despite the observed heterosis, pregnancies of within-strain matings were not more likely to be blocked than pregnancies of between-strain matings. A power analysis revealed that if we missed an existing effect (type-II error), the effect must be very small. If a female gave birth, the number and weight of newborns were not significantly influenced by the stimulus males. In conclusion, we found no support for the 'compatible-genes sexual selection hypotheses'.     

Ergocryptine and pregnancy maintenance in hamsters.

Ergocryptine (ECR) terminated pregnancy in hamsters when administered on Day 5; when ECR was given on Day 6 the response was diminished, and pregnancy continued after ECR treatment on Day 7. The abortifacient action of ECR in Day 5 pregnant hamsters was overcome by exogenous prolactin but not FSH and LH. When sera collected from hamsters on different days of gestation were examined for their ability to neutralize the effect of ECR in Day 5 pregnant hamsters, a peak of luteotrophic activity was observed in sera collected on Days 10 and 11. The results of these studies suggest that in hamsters the role of hypophyseal prolactin in luteal support is diminished by Day 7 of pregnancy, and the appearance of luteotrophic activity in sera collected on Days 10 and 11 may be indicative of a placental luteotrophin.     

Disruption of pair bonding induces pregnancy failure in newly mated mongolian gerbils (Meriones unguiculatus)

The efficacy of several exteroceptive factors including exposure to a 'strange' male, were tested to induce pregnancy failure in newly mated Mongolian gerbils. Only physical separation of monogamous pairs consistently induced pregnancy failure. This phenomenon was restricted to the immediate period after mating and is believed to be due to disruption of pair bonding, with associated luteal and implantation failure. Concurrent lactation annulled this effect.