Influence of embryonic and maternal genotype on gestational events in the mouse

Mice with high and low prenatal survival were used to study the influence of maternal and embryonic genotype on the timing of implantation, conceptus growth and gestation length. Mice selected for large litter size (Line S1) or rapid post-weaning weight gain (Line G) showed implantation was delayed and gestation prolonged in mice with low prenatal survival (Line G). Reciprocal transfer of Line-S1 and -G embryos to pseudopregnant recipients indicated that conceptus growth was influenced by maternal as well as embryonic genes, at least until mid-pregnancy. In contrast, fetal genotype had a major effect on the length of gestation.     

The effect of undernutrition on the establishment of pregnancy in the ewe

The relationship between nutrition and reproduction in sheep has been the subject of research in several international groups. This review will particularly focus on the effects of undernutrition on the potential causes of reproductive failure including abnormalities of the ovum or the embryo, luteal inadequacy and failure of the supply of progesterone to the uterus, or the mechanisms involved in maternal recognition of pregnancy. The level of nutrition and peripheral progesterone concentrations are inversely related, and increased rates of embryo loss, associated with higher progesterone concentrations in ewes with low levels of nutrition have been reported. Undernutrition may act through changes in the distribution of progesterone in the endometrium. Thus, lower endometrial levels on day 5 of the cycle in ewes fed half of their maintenance requirements have been observed, providing a link between the known role of progesterone in embryo survival by the modulation of uterine function and the higher embryo losses found in undernourished ewes. The evidence of an effect of maternal nutrition on IFNtau secretion from the conceptus and of PGF2alpha production from the uterus is presented. Moreover, undernutrition provokes a reduction in the sensitivity of the endometrium to progesterone that may affect embryo survival. Finally, a state of undernutrition induces changes in the endometrial sensitivity to steroid hormones at early stages of pregnancy that could adversely alter uterine environment to the detriment of embryo survival.     

Embryo survival, and fetal and placental growth following elevation of maternal estradiol blood concentrations in the rat.

High doses of estrogens cause embryonic mortality, and fetal and placental growth retardation in rats. This study addresses the physiological relevance of such findings. Estradiol benzoate (EB), by s.c. injection, or estradiol-17beta (E2), delivered by a miniosmotic pump, raised maternal E2 concentrations from only slightly above control values to 5-fold. EB (1 microgram/day) over Days 6-13, 8-13, and 11-13, and continuous infusion of E2 (15 ng/h; Days 10-13) reduced fetal survival to 0%, 0%, 22%, and 75%, respectively. Single injections of EB showed that its lethal effect declined rapidly over Days 9 (44% survival) to 13 (90% survival). Embryos died within 48 h, but death was not due to luteal failure since progesterone levels were maintained and progesterone administered with EB did not reduce mortality. Administration of EB at 1 microgram/day (Days 14-21) or E2 at 40 ng/h (Days 13-16) retarded fetal and placental growth but did not affect survival. The rat embryo is highly sensitive to elevated maternal estradiol concentrations over much of gestation. The early lethal effect implies that endogenous E2 production is carefully regulated to maintain pregnancy; the latter growth-retarding effect suggests that E2 may have a role in the normal control of fetal growth.     

Post-insemination milk progesterone concentration and embryo survival in dairy cows

Logistic regression analysis was used to evaluate the relationship between post-insemination milk progesterone concentration and embryo survival, and between milk yield and milk progesterone concentration. Milk samples were collected on Days 1, 4, 5, 6, and 7 (insemination=Day 0) following 871 inseminations in spring-calving dairy cows. Milk progesterone concentrations were measured by enzyme-immunoassay and pregnancy diagnosis was conducted with transrectal ultrasonography at approximately Day 30. There was a negative linear relationship (P<0.01) between milk progesterone concentration on Day 4 and embryo survival while, in contrast, there was a positive linear and quadratic relationship between milk progesterone concentration on Days 5, 6 and 7 (P<0.05) and also between the rate of change in progesterone concentrations between Days 4 and 7 inclusive and embryo survival (P<0.05). There was a weak negative linear relationship between average daily milk yield at the time of insemination and milk progesterone concentrations (P<0.001). There was no association between many production parameters, including liveweight and body condition score measured at various stages between calving and insemination, and milk progesterone concentration between Days 4 and 7 inclusive (P>0.05). In conclusion, low progesterone during Days 5-7 (after insemination) was associated with low fertility in dairy cows and there were indications of a range of progesterone concentrations within which embryo survival was maximal.     

Effect of progesterone supplementation in the first week post conception on embryo survival in beef heifers

Progesterone is essential for establishment and maintenance of pregnancy in mammals. The objective of this study was to examine the effect of elevating progesterone during the different physiological stages of early embryo development on embryo survival. Estrus was synchronized in cross-bred beef heifers (n = 197, ∼2-years old) and they were inseminated 12-18 h after estrus onset (=Day 0). Inseminated heifers were randomly assigned to 1 of 3 treatments: (1) Control, n = 69; (2) progesterone supplementation using a Controlled Internal Drug Release Device (CIDR) from Day 3 to 6.5, n = 64; or (3) progesterone supplementation using a CIDR from Day 4.5 to 8, n = 64. Body condition (BCS) and locomotion scores (scale of 1-5) were recorded for all animals. Animals with a locomotion score ≥4 (very lame) were excluded. Embryo survival rate was determined at slaughter on Day 25. Conceptus length and weight were recorded and the corpus luteum (CL) of all pregnant animals was dissected and weighed. Supplementation with exogenous progesterone increased (P < 0.05) peripheral progesterone concentrations, but did not affect embryo survival rate compared with controls. Mean CL weight, conceptus length and conceptus weight were not different between treatments. There was a positive relationship (P < 0.04) between the increase in progesterone concentrations from Days 3 to 6.5 and embryo survival rate in treated heifers and a similar trend existed between the increase from Days 4.5 to 8 (P < 0.06). There was also a positive relationship (P < 0.05) between the progesterone concentration on Day 6.5 and the embryo survival rate in treated heifers. A direct correlation was seen between locomotion score and embryo survival rate, with higher (P < 0.05) early embryo survival rates in heifers with a lower locomotion score. In conclusion, supplementation with progesterone at different stages of early embryo development increased peripheral progesterone concentration and resulted in a positive association between changes in progesterone concentration during the early luteal phase and embryo survival rate. Supplementation with progesterone had no effect on either CL weight or conceptus size in pregnant animals. Lameness had a significant negative effect on early embryo survival.     

Relationships between maternal hormone secretion and embryo development on day 5 of pregnancy in dairy cows

In cattle, increasing early embryonic losses are associated with inadequate progesterone concentrations within the first three weeks of pregnancy. The aim of this study was to investigate the complex relationship between early maternal progesterone concentration and embryo development early within the first week of pregnancy, specifically, on day 5 post-oestrus in dairy cows. Twenty Holstein-Friesian cows at the end of lactation were inseminated at oestrus (day 0) and on day 5 post-oestrus cows were slaughtered and the reproductive tract flushed to determine the presence and stage of embryo development. Three cows that had failed to synchronise correctly were excluded from analysis while in the remaining 17 cows 11 (65%) were pregnant with embryos at the morula (n = 3), 9-16 (n = 3) and 8-cell (n = 5) stages of development. No differences in day 5 plasma progesterone concentrations or corpus luteum (CL) size or progesterone content were observed between pregnant (n = 11) and non-pregnant (n = 6) cows. In cows with embryos beyond the 8-cell stage of development (n = 6) plasma progesterone concentration (P < 0.001) and CL weight (P < 0.01) were higher and plasma insulin concentrations lower (P < 0.001) than in cows with 8-cell embryos (n = 5). In addition there was a negative relationship between plasma progesterone and plasma insulin in pregnant cows (R(2) = 0.65; P < 0.005). In cows with an embryo present in the oviduct, oviductal glucose concentrations were lower (P < 0.05) than in cows with no embryo present. These results confirm progesterone is not only directly associated with embryo development, but that it may indirectly modulate embryo development via changes in the oviductal environment. In summary, the association between maternal progesterone concentration and embryo development exists as early as day 5 of pregnancy in dairy cows.     

Evidence of an association between the survival of embryos and the periovulatory plasma progesterone concentration in the ewe

Plasma progesterone concentration and embryo survival were determined during successive pregnancies in ewes throughout one breeding season. The probability of an embryo surviving was associated with the progesterone concentration on the days around ovulation, with the timing of the increase from periovulatory to luteal values, and with the rate at which progesterone concentrations increased. Individual embryo survival decreased both as the number of corpora lutea increased, and towards the end of the breeding season; the latter effect could be explained entirely by differences in progesterone concentration. Considerable variation in progesterone secretion and in embryo survival was observed within the same ewes during successive pregnancies. Such variability in progesterone concentrations during early pregnancy may be a cause of some embryo mortality.     

Associations between milk progesterone concentration on different days and with embryo survival during the early luteal phase in dairy cows

The relationships between the concentration of milk progesterone and early embryo survival on Days 4-8 inclusive and between the concentration of progesterone on different days from Days 0-8 inclusive following ovulation and insemination were examined in dairy cows. The relationships were examined following 77 randomly chosen artificial inseminations to cows in standing oestrus. There was a significant (P < 0.05) linear and quadratic relationship between the concentration of milk progesterone on each of Days 4-6 after ovulation and the probability of embryo survival. There was no association (P > 0.05) between milk progesterone concentration and probability of embryo survival on Days 7 and 8 after ovulation. There were no associations between milk progesterone concentration on Days 0-2 and the concentrations on Days 4-7, however, progesterone concentrations on Days 4 and 5 were highly predictive of the concentration on Days 6 and 7, respectively. Overall, the results indicate that suboptimal progesterone support during the early luteal phase is likely to deleteriously affect embryo viability and in addition, that it is possible to predict milk progesterone concentrations during the early luteal phase based on earlier stage concentrations and thus identify cows at risk of early embryo loss.     

Donor and recipient ewe factors affecting in vitro development and post-transfer survival of cultured sheep embryos

Donor and recipient factors were assessed during development of embryos following superovulation, collection at the pronuclear and two-cell stage, culture in Synthetic Oviduct Fluid medium for 5 days and twin transfer into synchronised recipients to elucidate what factors affect embryo development and post-transfer survival. In particular, the administration of exogenous progesterone to recipients using an intravaginal CIDR^TM device immediately following embryo transfer was investigated.

From 138 embryos collected from 30 donor ewes, 75% (103) were of transferable quality following culture, of which 100 were transferred to 50 recipients. There was significant variation (P < 0.001) in embryo development to the blastocyst stage between different donor ewes, but this was not related to the donor ovulation rate. At ultrasound sonography (approximately Day 60 of pregnancy), 58% of recipients were pregnant and 42% embryos had survived. Donor ovulation rate was related to embryo survival (P < 0.05) after transfer; the survival rate of embryos from ewes with high ovulation rates was lower than that of embryos from ewes with low ovulation rates. Exogenous progesterone supplementation following transfer did not affect embryo survival, rate of embryo development or plasma progesterone levels. In general, the results from this study suggest that factors other than efficacy of embryo culture can affect the outcome of embryo survival following transfer and that, where possible, these factors should be considered and balanced in experimental designs.     

Embryo survival and conceptus growth after reciprocal embryo transfer between Chinese Meishan and Landrace x Large White gilts

Embryos were transferred between Meishan and Landrace x Large White (control) gilts on Day 4 or 5 to establish approximately equal numbers of all four possible combinations of donor breed and recipient breed. The breed of the donor gilt significantly (P less than 0.01) affected embryo survival with 44.5% of transferred Meishan embryos and 69.6% of transferred control embryos surviving to Day 30 +/- 1. There was no influence of the breed of the recipient gilt on the proportion of embryos which survived. These differences in embryo survival between the two breeds could not be explained by differences in (1) the number of embryos transferred, (2) the stage of development of the embryos transferred, (3) the interval between ovulation and transfer or (4) the degree of asynchrony between donor and recipient gilt. On Day 30 +/- 1 embryos from control donors developed into longer fetuses (P less than 0.01) with larger allantoic sacs (P less than 0.05) than did embryos from Meishan donors. Fetuses in control recipients were longer (P less than 0.01), heavier (P less than 0.001) and had larger allantoic sacs (P less than 0.05) than fetuses occupying Meishan uteri. The interaction between breed of donor gilt and breed of recipient gilt did not significantly affect conceptus growth. These results suggest that Meishan pig embryos may be less tolerant to routine embryo transfer procedures than those of control gilts, that the genotype of the dam does not affect the proportion of embryos surviving to Day 30 +/- 1, and that both fetal and maternal factors affect conceptus growth.     

Genetic control of linolenic acid concentration in seed oil of rapeseed (Brassica napus L.)

Summary Results from a diallel mating of two rapeseed lines with distinctly different linolenic acid concentration show that this trait is mainly under control of nuclear genes of the embryo. However, significant differences in reciprocal F₁, BC₁ and BC₂ indicate maternal control, which is realized by interaction between maternal genotype and nuclear genes of the embryo. Additionally, temperature exerts considerable influence on the degree of maternal control. Since no reciprocal differences are detectable in F₂, cytoplasmic factors seem not to be involved in the inheritance of linolenic acid concentration. Hypotheses on the physiological nature of maternal control of this trait are discussed.     

Supplemental progesterone during early pregnancy exerts divergent responses on embryonic characteristics in sows and gilts

Progesterone (P4) plays a key role in pregnancy establishment and maintenance; during early pregnancy, P4 stimulates the production and release of uterine secretions necessary for conceptus growth prior to implantation; therefore, exogenous P4 supplementation may improve embryo development. This study evaluated the effects of supplementation during early pregnancy with long-acting injectable progesterone or altrenogest on embryonic characteristics of sows and gilts. Thus, a total of 32 sows and 16 gilts were used. On day 6 of pregnancy sows and gilts were allocated to one of the following groups: non-supplemented; supplemented with 20 mg of altrenogest, orally, from days 6 to 12 of pregnancy; supplemented with 2.15 mg/kg of long-acting injectable progesterone on day 6 of pregnancy. Animals were killed on day 28 of pregnancy, and ovulation rate, embryo survival, embryo weight, crown-to-rump length, uterine glandular epithelium and endometrial vascularization were assessed. Treatments had no effect on pregnancy rate, embryo survival or endometrial vascular density (P > 0.05). Non-supplemented gilts presented larger and heavier embryos compared to gilts from supplemented groups (P < 0.05). Sows in the altrenogest group presented larger and heavier embryos compared to non-supplemented sows and sows supplemented with long-acting injectable progesterone. In conclusion, supplementation of sows and gilts with progestagen from day 6 of pregnancy can be used as a means to improve embryo survival without deleterious effects.     

Imprinting of the MEDEA polycomb gene in the Arabidopsis endosperm.

In flowering plants, two cells are fertilized in the haploid female gametophyte. Egg and sperm nuclei fuse to form the embryo. A second sperm nucleus fuses with the central cell nucleus that replicates to generate the endosperm, which is a tissue that supports embryo development. MEDEA (MEA) encodes an Arabidopsis SET domain Polycomb protein. Inheritance of a maternal loss-of-function mea allele results in embryo abortion and prolonged endosperm production, irrespective of the genotype of the paternal allele. Thus, only the maternal wild-type MEA allele is required for proper embryo and endosperm development. To understand the molecular mechanism responsible for the parent-of-origin effects of mea mutations on seed development, we compared the expression of maternal and paternal MEA alleles in the progeny of crosses between two Arabidopsis ecotypes. Only the maternal MEA mRNA was detected in the endosperm from seeds at the torpedo stage and later. By contrast, expression of both maternal and paternal MEA alleles was observed in the embryo from seeds at the torpedo stage and later, in seedling, leaf, stem, and root. Thus, MEA is an imprinted gene that displays parent-of-origin-dependent monoallelic expression specifically in the endosperm. These results suggest that the embryo abortion observed in mutant mea seeds is due, at least in part, to a defect in endosperm function. Silencing of the paternal MEA allele in the endosperm and the phenotype of mutant mea seeds supports the parental conflict theory for the evolution of imprinting in plants and mammals.     

Use of fluorogestone acetate after breeding to reduce the effect of premature luteal regression in dairy goats when superovulation is induced with FSH

The aim of the present study was to determine the effect of fluorogestone acetate (FGA) administered after mating, on embryo production in the dairy goat subjected to conventional superovulatory and embryo recovery protocols. Adult does, most of them of the French Alpine breed, were randomly assigned after a FSH-superovulatory estrus and fertile matings to a control group (n=20) or to a treated group (n=20) in which intravaginal sponges impregnated with FGA were inserted after mating and remove before embryo collection (day 6). Blood samples were collected every 12h from days 1 to 7 post-estrus and serum progesterone concentrations were determined. The FGA-group had a lesser percentage of does with normal corpora lutea (CL) and a greater percentage of animals with CL in regression or mixed (normal and in regression) when compared with the control group (13.3 and 64.7%, 53.3 and 23.5%, and 33.3 and 11.8%, respectively; P<0.05). Mean number of normal CL per doe was less and mean number of regressed CL greater in FGA as compared with the control group (4.2 compared with 10.7 and 8.5 compared with 3.6, respectively; P<0.05). There were no differences (P>0.05) in recovery rate, total number of CL, total recovered structures, oocytes and transferable and non-transferable embryos between groups. Serum progesterone concentrations from day 5 to 7 post-estrus were lower (P<0.05) in FGA as compared with the control group. Percentage of does with luteal failure on day 6 post-estrus was greater in FGA as compared with the control group (86.6 compared with 33.3%; P<0.01). When considering only does with luteal failure on day 6 post-estrus, mean total recovered structures, transferable embryos and percentage of does rendering > or =3 transferable embryos were greater in the FGA compared with the control group (6.3 and 1.3 structures, 4.5 and 1.2 embryos, 67 and 17%, respectively; P<0.05). In does not having luteal failure, FGA administration did not appear to affect embryo production or embryo survival. These results indicate that FGA administration after mating improves embryo recovery in dairy goats with luteal failure after superovulatory treatment. However, it also increases the incidence of luteal regression when administered early in the estrous cycle.     

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.     

Influence of progesterone on oocyte quality and embryo development in cows

In cattle, the majority of embryo loss occurs very early during pregnancy (approximately Day 16), around or prior to maternal recognition of pregnancy. The actions of P4 in controlling LH pulsatility and ovarian follicular development may impinge negatively on oocyte quality. A considerable proportion of embryo loss may be attributable to inadequate circulating progesterone (P4) concentrations and the subsequent downstream consequences on endometrial gene expression and histotroph secretion into the uterine lumen. Conceptus growth and development require the action of P4 on the uterus to regulate endometrial function, including conceptus–maternal interactions, pregnancy recognition, and uterine receptivity for implantation. This review summarizes recent data highlighting the role of progesterone in determining oocyte quality and embryo development in cattle.     

Genetic control of survival of frozen mouse embryos

Lines of mice selected for increased litter size (L+), increased body weight (W+), or randomly (K) were used to study genetic variation in embryo cryosurvival in response to standard cryopreservation protocols. A total of 60528-cell embryos from 400 females were used in two studies. In Study 1, embryos from L+, W+, and K were frozen by slow-cool and ultrarapid (direct-plunge) methods to evaluate effects of selection on cryosurvival and genotype X freezing method interaction. Post-thaw survival (PTS) was measured as percentage of recovered embryos developing in vitro to blastocyst per donor female. Nonfrozen control embryos developed similarly for each line. Within slow-cool freezing, lines differed (W+ greater than K, W+ = L+, L+ = K; p less than 0.05); no differences were observed within the ultrarapid freezing. However, line X method interaction effects on PTS were not significant. In Study 2, reciprocal crosses were made between L+ and K and between W+ and K. Hybrid and pure line embryos were frozen by slow-cooling. Control embryos developed similarly for all genotypes. Selection lines did not differ for overall PTS. However, hybrid embryos from L+ dams were superior to those from K dams (84 vs. 61%; p less than .001). No overall embryo heterosis was observed. Differences were not significant among embryo genotypes or treatments for cell number or in vivo survival. These results demonstrate significant correlated responses in embryo post-thaw cryosurvival due to selection, and implicate both maternal and embryonic genomes as controlling mouse embryo cryosurvival.     

Embryo survival in dairy cows managed under pastoral conditions

Efficient pasture-based milk production systems require a compact calving pattern aligned to the onset of the grazing season, a 365-day calving interval and low culling rates for infertility. Achievement of these targets requires high herd reproductive performance. While high genetic merit Holstein cows produce more milk in grass-based systems their fertility is compromised. Management of the modern high genetic merit Holstein dairy cow presents a major challenge in pasture-based systems of production. It appears that the extent of early embryo loss is greater (up to 20% points greater) in the modern high-producing dairy cow and that a much higher proportion of the embryos die before day 7 following insemination in contrast to heifers and lower yielding cows. About 7-8% of pregnancies are lost between days 30 and 90 of gestation with no evidence that loss rate is related to cow genetic merit, parity or level of production. Systemic concentrations of progesterone during both the cycle preceding and following insemination affect embryo survival rate with evidence that too low or indeed too high a concentration of progesterone been negatively associated with embryo survival rate. Peripheral concentrations of both progesterone and oestradiol are lowered by increased plane of feed intake due to increased metabolic clearance rate of the steroids, which is related to liver blood flow. It appears that high producing dairy cows have an increased risk of embryo death as a result of lowered peripheral concentrations of progesterone as a consequence of increased hepatic metabolism of progesterone. Uterine expression of mRNA for progesterone receptor, oestradiol receptor and retinol binding protein mRNA appears to be sensitive to changes in peripheral concentrations of progesterone during the first week after AI. It would appear that energy balance and dry matter intake during the 4 weeks, immediately after calving are critically important in determining conception rate when cows are inseminated at 70-100 days post-calving. Concentrate supplementation of cows at pasture during the breeding period has minimal affects on conception rates though sudden reduction in dietary intake should be avoided. For pasture-based systems of milk production more balanced breeding strategies, with greater emphasis on fertility and feed intake must be developed.     

Effect of long-term selection for early postnatal growth rate on survival and prenatal development of transferred mouse embryos

Reciprocal embryo transfer procedures were performed among mouse selection lines to examine prenatal maternal effects on survival and development of transferred embryos. Mice were from generations 28 and 29 of an experiment to select for (i) increased body weight again from 0 to 10 days (E+); (ii) decreased body weight gain from 0 to 10 days (E-); or (iii) a randomly bred control line (C). A total of 118 embryo transfer procedures performed 12 h after conception resulted in 983 progeny born to 89 litters. There was a 39% overall embryo survival rate and 75% overall pregnancy success rate. Response to superovulation and oestrous synchronization was significantly lower (P < 0.01) in the E+ line. E+ individuals that did superovulate produced an average of 37 oocytes per flush, which was significantly higher than in the control line mice (29 oocytes per flush; P < 0.01). The ability to complete pregnancy successfully was not influenced by uterine environment or embryo-uterine interaction. In contrast, embryo survival in successful pregnancies was significantly affected by uterine environment. There were large maternal effects for body weight and tail length at birth; E+ recipients produced pups that were significantly larger than E- recipient pups (P < 0.01), which in turn were significantly larger than pups gestated by control recipients (P < 0.01).     

Changing responsiveness of luteal cells of the marmoset monkey (Callithrix jacchus) to luteotrophic and luteolytic agents during normal and conception cycles

Dispersed marmoset luteal cells were incubated for 2 h and progesterone production measured after exposure to hCG, cloprostenol, dibutyryl cAMP, PGF-2 alpha, PGF-2, adrenaline or melatonin. The cells were studied on Days 6, 14 and 20 after ovulation in conception and non-conception cycles. Luteal cells from Day 14 non-pregnant marmosets were compared with human luteal cells taken in the mid-luteal phase. All the treatments stimulated progesterone production including cloprostenol, which is luteolytic when administered to the marmoset in vivo, but the degree of response varied with the stage of the cycle or pregnancy and between marmoset and human luteal cells. In the marmoset, overall analysis of the effect of the treatments showed that, on Day 6 after ovulation, there was no significant effect of any of the treatments in cells from pregnant or non-pregnant animals. In contrast, luteal cells from non-pregnant animals on Day 14 showed a significant response to the treatments (F (8,41) = 2.79, P less than 0.0145) whereas cells from pregnant Day-14 animals were responsive; in cells from pregnant animals, the control production of progesterone was high and already equivalent to the levels stimulated by the treatments. By Day 20, cells from pregnant animals produced lower control concentrations of progesterone than did those on Day 14 and there was a significant overall effect of the treatments (F (8,33) = 3.78, P less than 0.003). These results show that the marmoset CL gains responsiveness to treatment between Days 6 and 14 after ovulation in the non-pregnant cycle. In pregnancy, on Day 14, 2 days after attachment of the embryo, the high control concentrations of progesterone and absence of response to treatment suggest that an embryo message may have affected the CL, providing an endogenous stimulus.