Participation of embryonic genotype in the pregnancy block phenomenon in mice.

Pregnancy block by male pheromones in mice differs in incidence depending on the combination of strains. Female mice of BALB/cA strain mated with BALB/cA males show a 100% pregnancy block when exposed to males of inbred strain DDK shortly after copulation (Chung et al., Biol Reprod 1997; 57:312-319). In the present study, BALB/cA females mated with the males of other strains--CBA/J, C3H/HeN, C57BL/6Cr, and IXBL--showed higher pregnancy rates (66.6-87. 5%) even when they were exposed to DDK males. In the pharmacological induction of pregnancy block with dopamine agonist (bromocriptine, 4 mg/kg BW), BALB/cA females mated with BALB/cA males showed a 100% pregnancy block. In contrast, BALB/cA females mated with CBA/J, C3H/HeN, and C57BL/6Cr males showed higher pregnancy rates (40-70%). These results suggest that the better pregnancy rate of BALB/cA females mated with alien males may be due to the stronger viability of F(1) embryos. This interpretation was confirmed by an embryo transfer experiment in which a higher implantation rate was observed when BALB/cA embryos grown in BALB/cA females exposed to BALB/cA males were transferred into recipient BALB/cA females exposed to DDK males. These results suggest that the embryonic genotype or viability of the embryo is one factor contributing to the occurrence of pregnancy block by male pheromones in mice.     

Pregnancy and early reproductive failure in the baboon

We documented normal pregnancy and the rate of pregnancy failure in female baboons by measuring chorionic gonadotropin (bCG) and progesterone (P) levels in 162 mated cycles of 70 baboon females on days 10, 12, and 14 postovulation. Females were mated with males during turgescene. The presence of pregnancy was defined by bCG levels >20 μg/ml by day 14 postovulation and/or documentation of a gestational sac using ultrasonography. Of the 162 cycles, 75 were fertile. Of these animals, 33 were used in other studies and thus were not included in these analyses. The analyses are based on 43 pregnancies from 91 cycles that were untreated throughout their gestations. Twenty-six of these pregnancies had abnormal bCG and/or progesterone levels in early pregnancy. All of those pregnancies with abnormal endocrine parameters terminated with spontaneous abortion (60%). Certain abnormal bCG patterns were repeatedly observed in some animals and were correlated with repeated spontaneous abortions. Of 17 pregnancies with normal bCG and P patterns, 15 (88%) continued to term with a normal fetal outcome. In this study, a pregnancy rate per mated cycle of 47% was observed, yet 60% of untreated pregnancies abortyed spontaneously. Overall 16% of the mated cycles had continuing pregnancies with normal outcome. These studies demonstrate that a high rate of early abortions occurs in the baboon and that a single bCG determination is insufficient to define the presence of a “normal” pregnancy which might be expected to carry to term with a normal outcome.     

Influence of single-sire and multiple-sire natural mating on pregnancy rate of beef heifers

Twelve mature Angus bulls possessing similar semen quality were used to study the effects of single and multiple services and single-sire and multiple-sire mating on pregnancy rate of beef heifers. Heifers were observed for natural estrus twice daily and mated within 6 hr of onset of estrus. Pregnancy rate was determined by rectal palpation at 60 days post-mating. Pregnancy rate of heifers receiving one service per estrus averaged 62.1% (n=124) and pregnancy rate of heifers receiving multiple services (single-sire) per estrus averaged 62.9% (n=105). Pregnancy rate per estrus of heifers mated to three sires (multiple-sire, one service per sire; 74.0%, n=123) was significantly higher than that of heifers mated to a single sire (62.9%). The range in pregnancy rate per bull for single-sire matings was large (0 to 95%), but this range was reduced among the same bulls used for multiple-sire matings (68 to 84%). These results indicate that pregnancy rate to one service did not differ (P>.10) from pregnancy rate to multiple services among single-sire matings, but multiple-sire matings increased (P<.05) pregnancy rate per estrus by 11% over that obtained with single-sire matings.     

Stimulation of alloreactive cytotoxic T cells by paternal major histocompatibility antigens during murine pregnancy

In an effort to elucidate T cell reactivity toward paternal major histocompatibility (MHC) antigens during pregnancy, the ability of pregnant mice to develop alloreactive cytotoxic T lymphocytes (CTL) was studied in individual multiparous females mated with MHC congeneic strains of B10 background. Spleen cells obtained from B10.BR females mated to allogeneic males manifested strikingly higher CTL than those from animals mated to syngeneic males or from virgins; syngeneically mated animals were equivalent to virgin controls in CTL responses. The augmented CTL response in allogeneic pregnancy was detected not only by stimulation with the paternal MHC antigens but also by an unrelated MHC haplotype. However, this augmentation was found only during pregnancy in that 2-5 days after the delivery the CTL activity in allopregnant animals returned to a level comparable to that of virgin controls. No suppressor cells were detected at this stage. These observations suggest that maternal T cells recognize MHC disparity with the fetus in some way during pregnancy. Anti-MHC antibodies, immunoglobulin (Ig) M, and IgGs of all subclasses were not detected in these animals throughout multiple pregnancies.     

Effect of progesterone,mifepristone, and estrogen treatment during early pregnancy on conceptus development and uterine capacity in Swine

A series of experiments was performed to investigate the influence of progesterone at Days 2 and 3 of pregnancy on conceptus development and uterine capacity. In experiment 1, unilaterally hysterectomized-ovariectomized (UHO) white crossbred gilts were given no treatment, estradiol valerate (5 mg given on Days 11 and 12), or progesterone (200 mg/day on Days 2 and 3 after mating). On Day 105 of pregnancy, each fetus and its associated placenta were weighed, and the number of live and dead fetuses was recorded for each litter. Early progesterone treatment reduced (P < 0.05) litter size (a measure of uterine capacity in UHO gilts). In experiment 2, intact white crossbred gilts were mated, given no treatment or progesterone treatment on Days 2 and 3 of pregnancy, and farrowed. Progesterone treatment decreased (P < 0.05) pregnancy rates. In pregnant gilts, progesterone had no effect on the number of live or stillborn piglets at birth, and gestation length was decreased (P < 0.05). Progesterone treatment did not affect the number of large or small piglets. In experiment 3, intact gilts were mated at estrus and then received 1). no treatment or treatment with 2). 100 mg, 3). 200 mg, or 4). 400 mg mifepristone (also known as RU486) on Day 2 of pregnancy. On Day 11 of pregnancy, both uterine horns were flushed, the number and diameter of each conceptus was recorded, and the flushed material was assayed for total protein and acid phosphatase. The 400 mg mifepristone treatment decreased conceptus diameter (P < 0.05) and total protein (P = 0.06) in the uterine flushings. In experiment 4, UHO gilts were mated at estrus, injected with either corn oil (control) or mifepristone (400 mg) on Day 2 of pregnancy, and killed on Day 105 of pregnancy, and the number and weight of live fetuses and placentas was recorded. In contrast to the effect of progesterone treatment, mifepristone decreased uterine capacity by decreasing the number of small conceptuses. These data suggest that progesterone concentrations on Days 2 and 3 of pregnancy in swine influence the rate of conceptus development during early pregnancy and uterine capacity during later pregnancy.     

Further evaluation of the pregnancy-linked down-regulation of the paternal antigen-specific splenic cytotoxic T lymphocyte activity in allogeneically pregnant mice.

Cytotoxic T lymphocyte (CTL) activity directed against paternal alloantigen was examined in allogeneically pregnant mice using various allogeneic combinations. The spleen cells from pregnant C57BL/6 (H-2b) mice mated with BALB/c (H-2d) male mice generated less anti-H-2d CTL after in vitro sensitization than those from unpregnant or syngeneically mated C57BL/6 mice. Different allogeneic combinations including the incompatibility at only D region of H-2 or minor histocompatibility loci were effective for downregulating the anti-paternal CTL activity in pregnancy. The downregulation of anti-paternal CTL activity induced by allogeneic pregnancy occurred at day 10 to day 18 of pregnancy, most extensively at day 14. The allogeneic pregnancy also downregulated the allogeneic CTL activities that had been amplified by injecting alloantigens before mating.     

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.     

Haemolymph ecdysteroid levels in female tsetse fly Glossina fuscipes (Diptera) during the first reproductive cycle: A comparison between virgin and mated females

The fluctuations in haemolymph ecdysteroid levels were recorded by radioimmunoassay during the first two ovarian cycles of the tsetse fly, Glossina fuscipes. During the first ovarian cycle, the patterns of ecdysteroid levels are similar in virgin and mated females. In virgins, the first oöcyte is not ovulated and ecdysteroid levels during the second vitellogenesis remain low. In mated females, vitellogenesis of the second oöcyte is concomitant with growth of the first larva in the mother's uterus and ecdysteroid levels are higher than during the first cycle. Two ecdysteroid increases were recorded in mated females and not in virgins. One coincides with vitellogenesis of the second oöcyte and the other occurs at the end of pregnancy and finishes after larviposition. The roles of ecdysteroids in the regulation of vitellogenesis, ovulation, pregnancy and larviposition are discussed.     

Effect of unilateral ovariectomy and subsequent ovum implantation in mice.

Unilateral ovariectomy (ULO) was done on any stage of the cycle and the animals were mated within day 1 to day 21 to observe the acute and long term effect of ULO on ovum implantation. Implantation reduced in proportion to single ovary if the animals were mated within 24 hr of ULO. Increase in ovarian weight along with an increase in implantation number continued in mated mice and reached at peak on day 19-21 of ULO (sacrificed after 6 days i.e., 25-27 days of ULO). After ULO the remaining ovary compensated within day 5-6 of ULO even during pregnancy. Ovarian histology showed stimulation of small antral follicles in mice mated on day 3 of ULO (sacrificed after 6 days i.e., day 9 of ULO) along with a decrease of large antral follicles and pre-antral follicles. Preantral follicles were at peak on day 12-14. Large antral follicles attained a peak on day 4 which slowly decreased. The occurrence of implantation in such ULO conditions are discussed.     

Induction of ovulation, mating, and conception in androgen-treated immature rats

Attempts were made to induce pregnancy in androgen-treated immature rats. Treatment with PMSG alone, which causes ovulation in normal immature rats, failed to cause ovulation in androgenized rats. However, treatment with PMSG plus LHRH was effective in causing ovulation. After ovulation, some of the normal and androgenized rats mated. Normal mated rats became pregnant but androgenized mated rats did not. However, when a pituitary gland was transplanted from a normal rat into the kidney capsule of an androgenized rat to maintain functional corpora lutea, implantation occurred in some of the mated animals. The positive decidual reaction in the uteri of such androgenized rats was similar to that observed in normal rats. These results suggest that the uterine sensitivity to blastocyst implantation of androgenized immature rats may be normal.     

Multi-male mating, probability of conception, and litter size in the prairie vole (Microtus ochrogaster)

We conducted a mating experiment in the laboratory using prairie voles, Microtus ochrogaster, to document that multi-male mating (MMM) can occur in this supposedly monogamous species and to test two hypotheses for the advantages of MMM in female mammals. The two hypotheses are that MMM (1) increases the probability of pregnancy and (2) increases litter size. We also tested the hypothesis that multiple copulations, rather than multiple partners, increases litter size and/or probability of pregnancy. Females were given a choice of mating with any of three males, each of which was tethered in a separate compartment. The mate choice bouts were recorded on videotape. We recorded the number of copulations and number of males with which females mated over a 24 h period. Litter size and probability of pregnancy were not significantly different for females that mated with one, two or three males. Increasing numbers of copulations, independent from the number of males, also did not increase litter size but did significantly increase the probability of pregnancy. MMM, at least in prairie voles, must serve some function other than increasing litter size and probability of conception.     

Relationship of body condition score and blood urea and ammonia to pregnancy in Italian Mediterranean buffaloes

The relationship of body condition score (BCS) and blood urea and ammonia to pregnancy outcome was examined in Italian Mediterranean Buffalo cows mated by AI. The study was conducted on 150 buffaloes at 145 +/- 83 days in milk that were fed a diet comprising 14.8% crude protein, 0.9 milk forage units.kg-1 dry matter and a non-structural carbohydrate/crude protein ratio of 2.14. The stage of the oestrous cycle was synchronised by the Ovsynch-TAI programme and blood urea and ammonia levels were assessed on the day of AI. Energy corrected milk (ECM) production and BCS were recorded bi-weekly. The pregnancy risk was 46.7% and was slightly lower in buffaloes with BCS < 6.0 and BCS > 7.5. There were no significant differences in ECM, urea and ammonia between pregnant and non-pregnant buffaloes. However, pregnancy outcome was higher (P = 0.02) in buffaloes with blood urea < 6.83 mmol.L-1. The likelihood of pregnancy for buffaloes with low urea blood level was 2.6 greater than for high urea level and exposure to a high urea level lowered the probability of pregnancy by about 0.25. The findings indicate that buffaloes are similar to cattle and increased blood levels of urea are associated with reduced fertility when animals are mated by AI.     

Plasma progesterone concentrations during early pregnancy in spring- and autumn-bred ewes

The objective of this experiment was to measure blood progesterone concentrations during early gestation to determine if the apparent reproductive failure in ewes bred out-of-season is due to a failure to conceive or embryonic loss. Blood samples were collected from spring- (n=61) and autumn-bred ewes (n=29) from Days 8 to 39 post-oestrus. Serum progesterone concentrations were analysed to ascertain whether ewes were ovulating and failing to maintain pregnancy, or conception was failing. Following pregnancy diagnosis 62 days after ram introduction, ewes were categorised as; no display of oestrus, mated but then identified as non-pregnant, or pregnant. A majority of spring-bred ewes that failed to display oestrus had silent oestrus (86%) and 66% of those ewes had abnormally short-lived corpora lutea. Circulating progesterone concentrations during dioestrus in ewes that had ovulated and displayed oestrus were unaffected by season. Similarly, progesterone concentrations during dioestrus did not differ between pregnant and mated non-pregnant ewes. The results indicated that while early luteylosis, low progesterone secretion from corpora lutea and embryo mortality did occur, these were in only a small proportion of ewes. Progesterone concentrations indicated that a majority of mated non-pregnant ewes had elevated progesterone concentrations necessary for the production of at least one viable embryo/foetus. This may be indicative to the failure of maternal recognition of pregnancy, and it is recommended that events surrounding this stage of pregnancy (Days 12-14) be examined more closely in ewes during the non-breeding season.     

An immunochemical demonstration of a pregnancy-specific protein in the horse and its use in the serological detection of early pregnancy

Two-dimensional crossed immunoelectrophoresis of sera from pregnant and non-pregnant horses, using antisera developed against early pregnant mare serum, revealed the presence of two immunologically related proteins one of which appeared to be specific to the pregnant state. This pregnancy-specific protein had beta 2-electrophoretic mobility and was first detectable at Day 6 after successful mating with a stallion. The second protein had gamma 2-electrophoretic mobility and was present in sera from pregnant and non-pregnant horses. The proteins were termed beta 2-horse pregnancy protein and gamma 2-horse protein respectively. The latter appeared to be immunologically related to the former in that the precipitin lines of the 2 proteins showed continuity. Samples from 16 mares mated with a stallion were investigated for the beta 2-protein during the first 3 weeks after mating. Of the 11 successful matings, confirmed by ultrasonic scanning at 90 days or by a successful outcome, 10 mares showed the presence of the protein. In all of 14 non-pregnant sera taken from mares not recently mated, the protein was not detectable. The validity of detection of beta 2-protein as an indication of pregnancy was clinically significant at the 10% level. The presence of the protein in 2 out of the 5 recently mated mares that did not become pregnant may be indicative of a biochemical pregnancy that failed at a later stage of gestation.     

Early pregnancy diagnosis by the ultrasonographical device and observation of fatal growth in cynomolgus monkeys (Macaca fascicularis)

Usefulness of the ultrasonographical device (USD) for early diagnosis of pregnancy and for observation of fetal growth was examined in the cynomolgus monkey (Macaca fascicularis). Firstly, 50 females, each of which was mated for 24 hours in an individual cage, were used. They were diagnosed for pregnancy by the use of USD 15 to 29 days and 35 days after mating. Then, 20 of them were judged to be pregnant. The early pregnancy diagnosis was based on the detection of gestational sac (GS) in the uterus. The GS was detected 20.0 +/- 1.8 days after mating. The duration from the day on which GS was first detected to the day on which the heart beat of embryo (HBE) was detected averaged 5.6 +/- 1.5 days. In 17 of these 20 pregnancies, GS was detected 17 to 23 days after mating. As regards the remaining three pregnant cases, GS and HBE were simultaneously detected on the 28th day of pregnancy in two cases, and on the 35th day in one case. Secondly, 225 females who were mated by the routine 3 day-mating system were used. Pregnancy diagnosis by USD was conducted on the 21st, 28th and 35th day after mating. Ninety-six of the 225 females were judged to be pregnant. In 33 (34.1%) of the 96 pregnant cases, GS was detected on the 21st day of pregnancy. In 57 females (59.4%), GS was detected on the 35th day. Furthermore, it was able to judge conditions of fetal growth by measuring the size of GS and biparietal diameter.(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

Pregnancy detection from fecal progestin concentrations in the red panda (Ailures fulgens fulgens)

The reproductive physiology of red pandas (Ailures fulgens fulgens) has not been well documented. This critically endangered species is not self‐sustaining in captivity despite several breeding populations, with low reproductive success and high infant mortality being leading causes of the decline. Hormone profiles were monitored in three groups of females (mated with birth, mated no birth, and not paired) to document pregnancy and parturition. Fecal samples were analyzed for progestins using a radio‐immuno assay. Females that gave birth had significantly higher progestins during the study period compared to females that mated and did not give birth and females that were not paired with a male. Two critical time frames were detected, Weeks 7–11 and Weeks 13–20, in which pregnant females could be differentiated from the others with a 95% confidence interval (CI). Detecting pregnancy in captive red pandas may assist animal care staff in management of the females and increase the survival rate of offspring. Zoo Biol 0:1–11, 2005. © 2005 Wiley‐Liss, Inc.     

Approaches to pregnancy diagnosis in the sable (Martes zibellina, Mustelidae, Carnivora) by noninvasive methods: Postimplantation period

To develop a reliable approach to pregnancy diagnosis in sables based on noninvasive methods of hormonal status assessment, the concentrations of immunoreactive compounds (IRC) binding with antibodies to progesterone have been measured in the feces of females at different stages of the reproductive cycle. The results show that this concentration is higher in truly pregnant than in mated but nonreproducing females. An increase in the IRC concentration relative to its individual baseline level may be regarded as a reliable indication of true 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'.     

Influence of male gonadal hormones and familiarity on pregnancy interruption in prairie voles

Pregnancy interruption (PI) was examined in female prairie voles, Microtus ochrogaster, exposed to stimuli from males 7 to 12 days after pairing. Urine from unfamiliar males interrupted pregnancy when placed directly on the external nares of newly mated females, but urine from familiar stud males was without effect. Castration of males did not reduce the efficacy of unfamiliar male urine in interrupting pregnancy. The neuroendocrine system of female prairie voles responded selectively to male urine as a function of its familiarity; the efficacy of male stimuli leading to PI was not dependent on gonadal hormones.