Effects of simulated high altitude (3800 m) on reproductive function in the pregnant rat.

Ovarian appearance, uterine contents (including numbers of fetuses, size and weight), and levels of gonadotropic hormones were studied in pregnant rats exposed to simulated altitude (3800 m). Comparisons were made at 11, 20 and 21 days of gestation between sea level controls rats bred at sea level and placed at reduced barometric pressure on confirmation of pregnancy (SHA). Although fewer fetuses were present in SHA rats on day 11 of gestation, litter size at days 20 and 21 was similar to controls. Body weight and length of SHA fetuses were significantly reduced by day 21 of gestation while placental weights remained similar to controls. The follicle-stimulating hormone (FSH) content in the pituitaries of SHA rats was significantly reduced at days 11 and 21 of pregnancy as compared with controls; luteinizing hormone (LH) content was lower at day 20 than at day 11 among SHA rats. Plasma gonadotropin levels were similar between SHA and control rats all all three stages of gestation. It is suggested that, despite lack of placental adaptation and altered endocrine functions, exposure of rats to hypoxic conditions only during gestation permits better maintenance of pregnancy than reported in rats with longer exposure to high altitude.     

Effects of preconceptional and gestational exposure to Tordon 202c on fetal growth and development in CD-1 mice

Female CD-1 mice were exposed to Tordon 202c (a picloram and 2,4-D combination herbicide) in the drinking water at concentrations of 0.21, 0.42, and 0.84% for 60 days prior to mating with untreated males. One-half of the pregnant females subsequently continued treatment throughout gestation while the remaining females were maintained on distilled water. Fetal weight, crown-rump length, placental weight, and maternal gestational weight gain were reduced in a dose-dependent manner following combined preconceptional and gestational exposure. The incidence of malformed fetuses (cleft palate, renal agenesis, hydronephrosis, unilateral testicular agenesis, and umbilical hernia) and fetuses with variants (especially incomplete ossification of the skeleton) were increased in a dose-dependent manner following combined exposure. Increased maternal mortality and decreased preconception weight gain were observed in the highest-dosage group. Relative maternal liver weight was increased in a dose-dependent manner. The results suggest that combined preconceptional and gestational exposure to Tordon 202c is required for teratogenesis and fetal growth depression. Preconceptional exposure alone is not effective in increasing the risk for embryotoxicity.     

Effects of pulsed magnetic fields on the developing mouse embryo

The influence of a pulsed magnetic field (PMF; sawtooth with 45-μs linear rise time and 5-μs decay time, peak strength of 15 μT, and frequency 20 pps) on the embryogenesis of CBA/S mice was investigated in five experiments based on a total of 707 exposed and 543 unexposed primigravidas. Sham and PMF exposures began on day 1 of gestation (experiments 1 and 2), on day 2 (experiment 3), on day 5 (experiment 4). and on day 7 (experiment 5): all exposures continued until day 19 post conception (p.c.) when they were terminated, at which time the following variables were measured: number of implants; number of placental resorptions; number of living fetuses; number of dead fetuses; number of malformations in living and dead fetuses; and length and body mass of living fetuses. Control dams were sham-exposed concurrently with corresponding. PMF-exposed dams. With the exception of experiment 5, in which exposure to PMF started on day 7 p.c., all groups of exposed mice had significantly more placental resorptions when compared with concurrent controls. The increased resorption rate was not reflected in a reduction in litter size or in the number of litters. A significant increase in malformed fetuses was not seen in any of the exposed groups, or when groups were pooled. Only in experiment 1 was the number of dead fetuses affected by exposure to PMF. The effect of PMF on the implantation rate was not significant. Body mass and length of exposed fetuses were significantly reduced only when the PMF treatment began on day 7 p.c. That PMF-treated mice had significantly more placental resorptions when exposure began on day 5 p.c. or earlier (before implantation), but not when exposure began on day 7 (after implantation), may indicate a causative pre-implantation effect. Because a PMF-induced increase in the number of resorptions has not been observed in other strains of mice, the effect might be strain-related. © 1993 Wiley-Liss, Inc.     

Intrauterine growth of the guinea pig fetal-placental unit throughout pregnancy: regulation by utero-placental blood flow

The relationships between uterine blood flow (UBF) and growth related changes in guinea pig placental weight, fetal weight, and crown-rump (C-R) length were examined at 5-day intervals throughout pregnancy. Between days 5 and 20 (Day 0 = ovulation) of pregnancy, UBF reached peak levels that were related to increases in total uterine weight as a result of fetal-placental differentiation. Between days 20 and 40 of pregnancy, UBF remained at constant, basal levels while placental weight increased at a rate of approximately 500 mg per 5-day interval. During this time period, fetal weight and C-R length continued to increase, but at a less rapid rate. Between days 40 and 55 of pregnancy, a secondary elevation in UBF was observed that was temporally related to third trimester growth of the fetus (ie, weight and C-R length). Near term (day 65), UBF decreased and placental growth reached a plateau, whereas fetal weight and C-R length continued to increase. In order to determine the direct effects of UBF on regulating fetal-placental growth, mid-gestational (days 35-50) pregnant guinea pigs were subjected to UBF reduction and the growth parameters of the manipulated fetal-placental units were then examined at 2, 3, 4, and 10 days post-alteration and compared to controls. The reduction in UBF at hyperemic pregnancy sites to basal flow levels resulted in an abrupt curtailment of subsequent placental growth noticeable within 48 hr of the vascular reduction. Subsequently, fetal weight and crown-rump length parameters were depressed with respect to controls, but not until 4 and 10 days post-alteration, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Fetal development, and placental and maternal plasma concentrations of progesterone in the little brown bat (Myotis lucifugus)

Progesterone concentrations measured in plasma samples from 280 bats captured during pregnancy or early lactation were related to fetal attributes indicative of stage of pregnancy. Fetal weight increased exponentially from 40 mg at crown-rump length of 6 mm to 2000 mg at 23 mm (term). Fetal weights at term accounted for up to 35% of the weight of intact pregnant animals. Progesterone concentrations increased from less than 5 ng/ml at 2 mm estimated crown-rump length to plateau values of approximately 65 ng/ml (geometric means) from 16 mm crown-rump length until the most advanced stages of pregnancy. Mean concentration in 8 post-partum bats, most of which were actively lactating, was 8.4 ng/ml; 11.6 ng/ml was measured in one animal that was carrying a wet neonate when sampled yet was still pregnant when captured 5 h earlier. Placental concentrations of progesterone ranged from 43 to 964 ng/g wet weight of tissue and mean values increased in a similar fashion though were about 4-fold greater than changes in plasma concentrations of the steroid. The concentrations in placental tissue were at least 15- to 20-fold higher than could be expected from blood contamination, indicating that placental steroidogenesis is likely to occur in this species.     

Chain elongation and desaturation of palmitic acid in liver microsomes of rats subjected to hyperbaric exposure.

The enzyme activities associated with chain elongation and desaturation of fatty acid in hepatic microsomes from rats held at 1 ATA of air, 1 ATA of He-O2, and 20 ATA of He-O2 were studied. It was found that both the microsomal chain elongation and desaturation of fatty acids were depressed in rats held at 1 ATA of He-O2 as compared to animals held at 1 ATA of air. When animals were exposed to an environment of 20 ATA of He-O2, the chain elongation of fatty acid was about the same as for rats held at 1 ATA of air and was two times greater than for the rats held at 1 ATA of He-O2. The desaturase activity was depressed as compared to the two groups of control animals held at 1 ATA of air and 1 ATA of He-O2.     

Effects of paternal subacute exposure to Tordon 202c on fetal growth and development in CD-1 mice

Male CD-1 mice were exposed to Tordon 202c (a picloram and 2,4-D combination herbicide) in the drinking water at concentrations of 0.21, 0.42, and 0.84% solutions for 60 days prior to mating with untreated females. Subsequently there was no exposure to Tordon 202c during gestation. Fetal weight and crown-rump length were reduced in the highest dosage group. The incidence of malformed fetuses (e.g., ablepharon, cleft palate, and unilateral agenesis of the testes) was increased in the middle dosage group while the incidence of fetuses with variants was increased in the lowest (e.g., an extra pair of ribs) and the highest dosage groups (e.g., incomplete ossification of the skeleton). The frequency of pregnancy failure was increased in the middle dosage group. Indices of paternal toxicity included increased lethality and decreased water consumption in the highest dosage group and increased relative spleen weights in the lowest and middle dosage groups. The results suggest paternally mediated reproductive toxicity.     

Non-invasive ultrasonographic characterization of phenotypic changes during embryo development in non-anesthetized mice of different genotypes

This study aimed to determine phenotypic changes during embryo development in the mouse, and the influence of genetic background, by non-invasive real-time ultrasonography. Serial scannings were performed from Day 4.5 after the appearance of the vaginal plug in a total of 34 adult mice of different strains (9BALB/c, 10C57BL/6 and 15 CD1). Embryonic vesicle diameter was measurable from Day 4.5 of pregnancy. Images of embryos were obtainable from Day 5.5 of gestation onwards, and crown-rump length and trunk parameters (diameter and area) were measured. At more advanced stages, the positions of fetuses prevented accurate measurement of crown-rump length; therefore, head diameters (occipito-snout length and biparietal diameter) were used as an alternative index of fetal size. All measurements correlated highly with gestational age (P<0.0005). No significant differences were observed between genotypes in early pregnancy, but during the last week of gestation trunk measurements were larger in CD1 embryos (P<0.05) while head diameters were larger in C57BL/6 conceptuses (P<0.05). There was a significant effect of genetic background on heart rate throughout pregnancy; although heart rate was similar in CD1 and C57BL/6 embryos (154.5+/-2.8 and 147.8+/-4.5 beats/min, respectively), it was significantly lower in BALB/c mice (127.0+/-2.1; P<0.005 vs. CD1 and C57BL/6).     

Maternal LPS Exposure during Pregnancy Impairs Testicular Development,Steroidogenesis and Spermatogenesis in Male Offspring

Lipopolysaccharide (LPS) is associated with adverse developmental outcomes including embryonic resorption, fetal death, congenital teratogenesis and fetal growth retardation. Here, we explored the effects of maternal LPS exposure during pregnancy on testicular development, steroidogenesis and spermatogenesis in male offspring. The pregnant mice were intraperitoneally injected with LPS (50 µg/kg) daily from gestational day (GD) 13 to GD 17. At fetal period, a significant decrease in body weight and abnormal Leydig cell aggregations were observed in males whose mothers were exposed to LPS during pregnancy. At postnatal day (PND) 26, anogenital distance (AGD), a sensitive index of altered androgen action, was markedly reduced in male pups whose mothers were exposed to LPS daily from GD13 to GD 17. At PND35, the weight of testes, prostates and seminal vesicles, and serum testosterone (T) level were significantly decreased in LPS-treated male pups. At adulthood, the number of sperm was significantly decreased in male offspring whose mothers were exposed to LPS on GD 13–17. Maternal LPS exposure during gestation obviously diminished the percent of seminiferous tubules in stages I–VI, increased the percent of seminiferous tubules in stages IX–XII, and caused massive sloughing of germ cells in seminiferous tubules in mouse testes. Moreover, maternal LPS exposure significantly reduced serum T level in male mice whose mothers were exposed to LPS challenge during pregnancy. Taken together, these results suggest that maternal LPS exposure during pregnancy disrupts T production. The decreased T synthesis might be associated with LPS-induced impairments for spermatogenesis in male offspring.     

The effects of subnutrition on the components of the gravid uterus in the doe

The effects of subnutrition on the caprine fetus and the other products of pregnancy were investigated in does. Two groups of does were fed on rations calculated to provide 100 and 25% of their energy and protein requirements for maintenance from 19 days before mating until 60 days after mating. Estrus was synchronized in does using PGF(2x). Approximately 60 days after natural mating, pregnant does were slaughtered, and the products of pregnancy were measured. Fetuses from the feed-restricted group were significantly lighter (P<0.05), had shorter crown-rump length (P<0.05), and the uterus contained a smaller volume of fetal fluids (P<0.02). Curved crown-rump length tended to be shorter and fetal placental membranes and cotyledons tended to be lighter (P<0.1) in the feed-restricted group. No significant difference was found between the 2 groups in the head length, number of placentomes, and weight of empty uterus. The number of fetuses affected the number of placentomes (P<0.001), weight of empty uterus (P<0.001), mass of total fetal fluids (P<0.001) and weight of ovaries (P<0.05), but not fetal measurements. Gestation length was found to significantly (P<0.001) affect all the fetal measurements but none of the placental measurements except for the total weight of cotyledons (p<0.001). The results of the study demonstrated detrimental effects of underfeeding on the caprine fetus and placenta.     

Effect of 71 ATA He-O2 on organ blood flow in the rat

Cardiac output and organ blood flow to major organs were investigated in awake rats at 1 atmosphere absolute (ATA) air and at 71 ATA He-O2. Radioactively labeled microspheres [15 +/- 1 (SD) micron] were injected into the left ventricle during constant-rate arterial blood sampling at 1 ATA air and subsequently at 71 ATA He-O2. Intra-arterial blood pressure was continuously recorded. The partial pressure of O2 was kept between 0.4 and 0.6 ATA. The results indicate that the mean blood pressure, heart rate, cardiac output, and organ blood flow are essentially unaltered in the rat at 71 ATA except for increased blood flow to the liver (122%, P less than 0.05), whereas the blood flow to the adrenals, the diaphragm, and the leg muscle fell (P less than 0.05).     

Effect of exposure to extremely low electro-magnetic field during prenatal period on mice spleen

Total body weight of newborns, the volume of spleen, and the number of megakaryocytes decreased following the exposure to ELF-MF (6 x 10(-3) T and 50 Hz) at 1-5, 6-10, 11-15, and 16-20 days of pregnancy of mice. The complete period of gestation was sensitive to ELF-MF exposure; the initial days were more prone to exposure. The results suggest that the use of ELF-MF producing instruments should be limited during gestation.     

Ultrasound fetal measurements and pregnancy associated glycoprotein secretion in early pregnancy in cattle recipients carrying somatic clones

Somatic cloning in the bovine species leads to high levels of fetal losses which occur throughout pregnancy. These losses are most often associated with fetal overgrowth, a syndrome known as large offspring syndrome (LOS), and excessive maternal plasma pregnancy serum protein 60 (PSP60), a protein similar to a pregnancy-associated glycoprotein of 67 kDa (PAG I67) produced by the bovine placenta. Predicting the outcome of pregnancies initiated from cloned embryos has become an important issue both to prevent potential harm to the mother because of excessive fetal size at birth and also to get a better understanding of the relationships between growth, differentiation and placental functions in developing cloned fetuses. Here, we report on a systematic analysis of fetal and placental development in the first trimester of pregnancy performed by ultrasonographic imaging and by measurement of the maternal concentrations of pregnancy associated glycoproteins (PAGS), using four different radioimmunoassays (RIA) (two homologous RIA systems with PSP60 and PAG I67; two heterologous RIA systems with PAG I67 as standard and tracer, and antisera anti-caprine PAGs). We showed that crown-rump length (CRL) in clones appeared smaller than controls at 35, 50 and 62 days (P<0.05). At 62 days of pregnancy, CRL in cloned fetuses that died before 90 days was smaller compared to the other cloned fetuses (P<0.05) whereas the width of the fetal sack and the biparietal diameter (BPD) was larger in fetuses that developed LOS in late gestation (P<0.05). Maternal PAGs concentrations were statistically different between controls and all clone recipients as early as Day 34, suggesting early abnormal placental glycoprotein synthesis for clone pregnancies regardless of pregnancy outcome. This work provides a practical, non-invasive tool to follow up clone pregnancies and suggests that primary growth retardation and abnormal placental function precedes excessive fetal and placental growth at later stages of pregnancy.     

Effects of low-frequency magnetic fields on fetal development in CBA/Ca mice

Effects of alternating magnetic fields (MFs) on the embryonic and fetal development in CBA/Ca mice were studied. Mated females were exposed continuously to a sinusoidal 50 Hz (13 μT or 0.13 mT root mean square) or a sawtooth 20 kHz (15 μT peak-to-peak) MF from day 0 to day 18 of pregnancy for 24 h/day until necropsied on day 18. Control animals were kept under the same conditions without the MF. MFs did not cause maternal toxicity. No adverse effects were seen in maternal hematology and the frequency of micronuclei in maternal bone marrow erythrocytes did not change. The MFs did not increase the number of resorptions or fetuses with major or minor malformations in any exposure group. The mean number of implantations and living fetuses per litter were similar in all groups. The corrected weight gain (weight gain without uterine content) of dams, pregnancy rates, incidences of resorptions and late fetal deaths, and fetal body weights were similar in all groups. There was, however, a statistically significant increase in the incidence of fetuses with at least three skeletal variations in all groups exposed to MFs. In conclusion, the 50 Hz or 20 kHz MFs did not increase incidences of malformations or resorptions in CBA/Ca mice, but increased skeletal variations consistently in all exposure groups. Bioelectromagnetics 19:477–485, 1998. © 1998 Wiley-Liss, Inc.     

Effects of secondary bile acids on the intrauterine development in rats

The effects of secondary bile acids (lithocholic--LCA, and deoxycholic--DCA) on the in vivo development of rat embryos and fetuses were studied. Daily intraperitoneal injections of 2 ml of 1 mM LCA and of 5 mM DCA during days 6 till 15 of pregnancy resulted in an increase of resorptions among 20 day-old fetuses to 22.8% and 9.9%, respectively, vs. 6.2% in controls. Similar injections on days 12 to 19 resulted in an increase of resorptions to 10.3% after treatment with LCA and to 36% after treatment with DCA. Percent of retarded embryos was similar for both bile acids: 7.7 and 8.7% after injections on days 6-15 and 12.3-12.5% after injections on days 12-19 of gestation. This was accompanied by a significant increase in the wet weight of the placenta of living embryos. Intraamniotic injections of 2 microliters of 1 mM LCA into 10 day-old embryos resulted in 18.5% resorptions (vs. 7.5% in controls), 9.2% malformations, and 3.1% growth retardations observed on day 12 of pregnancy. The rate of resorptions following this treatment increased on day 20 of pregnancy to 71% vs. 16% in controls. No differences were found in the wet weight of 20 day-old living fetuses or their livers and placentas between experimental and control groups following i.p. or intraamniotic injections. In addition, single intrauterine instillation of 0.2 ml of 1 mM LCA 10-14 days before mating with normal isogeneic males resulted in 9% of malformations among 12 day-old embryos while malformations were absent in the saline-injected controls. The deleterious effects of secondary bile acids to the embryos were accompanied by damage to the visceral yolk sac. These findings may be significant in relation to the complications previously associated with cholestasis of pregnancy in humans.     

Hyperbaric-induced subfertility in mice: lack of effect of decompression

Male mice were exposed to 50 atmospheres absolute (ATA). He for 30 min, twice weekly for 5 wk and their fertility assessed by subsequently mating with untreated virgin females. The exposure schedule was designed to provide compression and decompression times the same as previous positive studies of hyperbaric-induced subfertility but with much shorter periods at maximum pressure (30 min vs. 24 h). We postulated that, if subfertility were associated with exposure to pressure per se rather than with compression or decompression, then the present experiments would fail to produce a decrease in male fertility. The data were compared with those of a control group placed in the hyperbaric chamber at 1 ATA. The pregnancy rate for exposed vs. control mice was 89 vs. 86%, and the mean live liter size was 6.2 vs. 5.6. There were no statistically significant differences between these and other indexes of male fertility, and the results support our hypothesis that the hyperbaric-induced subfertility in male mice is not associated with these decompression procedures.     

Normal prenatal growth of the golden hamster (author's transl)]

In the golden hamster (Mesocricetus auratus) the gestation period and the loculus size of the gravid uterus from day 6 to day 15.5 (parturition) of gestation as well as the weight, width (umbilcus-black) and lenght (crown-rump) of the embryos from day 9 to the parturition were measured. The results obtained were as follows: 1. The gestation period of the golden hamster was 15 days and 15 +/- 3 hours post coitum. 2. The average values of the length (in the direction of uterine long axis) and width (mesome-trial-antimesometrial axis) in the loculus of the gravid uterus were 0.39 cm and 0.56 cm at 6 days, and 2.42 cm and 1.74 cm at 15.5 days (partiurition), respectively. 3. The average values of the crown-rump measurements, width (umblicus-back) and weight in the hamster embryos were 0.42 cm, 0.24 cm and 0.026 g at 9 days, and 2.40 cm, 1.39 cm and 2.272 g at 15.5 days (parturition), respectively. 4. The loculus size of the gravid uterus, the crown-rump measurements and width (umblicus-back) of the embryos in the period from day 13 to day 14 of gestation were markedly increased in comparison with any other period, and the weight increase of the embryos occurred rapidly from day 13 of gestation. 5. The shape of the loculus during the gestation was ovoid (mesometrial-antimesometrial axis) until the end of 10 days converged to the spherical form and thereafter changed gradually to the ovoid from in the direction of uterine long axis contraly to the previous days.     

Supplementation with high-dose cholecalciferol throughout pregnancy induces fetal growth restriction through inhibiting placental proliferation and trophoblast epithelial-mesenchymal transition

Vitamin D deficiency has been associated with adverse pregnant outcomes. Several studies investigated the effects of maternal vitamin D3 supplementation on fetal development with inconsistent results. The aim of this study was to investigate the effects of maternal supplementation with different doses of vitamin D3 on fetal development. Pregnant mice were administered with different doses of cholecalciferol (0, 2,000, 10,000, 40,000 IU/kg/day) by gavage throughout pregnancy. Fetal weight and crown-rump length were measured. Placental proliferation and mesenchymal characteristics were detected. HTR-8/SVneo cells were incubated in the absence or presence of calcitriol (500 nmol/L) to evaluate the effects of active vitamin D3 on migration and invasion of human trophoblast cells. Although a low dose of cholecalciferol was safe, fetal weight and crown-rump length were decreased in dams treated with high-dose cholecalciferol throughout pregnancy. Placental weight and labyrinth thickness were reduced in mice administered with high-dose cholecalciferol. An obvious calcification was observed in placentae of mice administered with high-dose cholecalciferol. Ki67-positive cells, a marker of placental proliferation, were reduced in mice administered with high-dose cholecalciferol. N-cadherin and vimentin, two mesenchymal markers, were decreased in cholecalciferol-treated mouse placentae and calcitriol-treated human trophoblast cells. MMP-2 and MMP-9, two matrix metalloproteinases, were downregulated in cholecalciferol-treated mouse placentae and calcitriol-treated human trophoblast cells. In addition, trophoblast migration and invasion were suppressed by calcitriol. Supplementation with high-dose cholecalciferol induces fetal growth restriction partially through inhibiting placental proliferation and trophoblast epithelial-mesenchymal transition.