Reproduction and development in rats chronologically exposed to 60-Hz electric fields

Previous studies have raised the possibility of reproductive and developmental changes in miniature swine chronically exposed to a strong 60-Hz electric field. Two replicate experiments on rats were performed to determine if similar changes could be detected in animals exposed under a comparable regime, which was based on average, induced-current densities and on the chronology of reproductive development, as dosimetrically and biologically scaled. Beginning at three months of age, female rats of the F0 generation and their subsequent offspring were chronically exposed to a 60-Hz electric field (100 kV/m unperturbed) for 19 h/day for the duration of experimentation. After four weeks of exposure, F0 female rats were mated to unexposed male rats during the field-off period. No significant developmental effects were detected in their litters, confirming our previous results with swine and rats. The F0 females were mated for a second time at 7.2 months of age, and the fetuses were evaluated shortly before term. In the first experiments, the incidence of intrauterine mortality was significantly less in exposed than in sham-exposed litters, and there was a tendency (P = .12) for an increased incidence of malformed fetuses in exposed litters. Neither end point was significantly affected in the second experiment. Copulatory behavior of the female F1 offspring, which were bred at three months of age, was not affected in either experiment. There was a statistically significant decrease in the fertility of F1 exposed females and a significant increase in the fraction of exposed litters with malformed fetuses in the first experiment; both end points were essentially the same in the sham and exposed groups of the second experiment. That the significant effects detected in the first experiment were not seen in the second may be attributed to random or biological variation. Alternatively, the finding may indicate that the response threshold for induction of malformations lies near 100 kV/m.     

Progeny of somatic cell nuclear transfer (SCNT) pig clones are phenotypically similar to non-cloned pigs

Systematic studies of cloned animals generated from adult somatic cell nuclei are critical in assessing the utility of somatic cell cloning in various applications, including the safety of food products from cloned animals and their offspring. Previously, we compared somatic cell derived cloned pigs with naturally bred control pigs on a series of physiological and genetic parameters. We have extended our studies to the F1 progeny of these clones to see whether these phenotypic differences are transmitted to the next generation. There were no differences in the average litter size between litters from cloned gilts and naturally bred controls (7.78 +/- 2.6 and 7.40 +/- 3.0, respectively; mean +/- SD) or in the degree of litter size variation (coefficients of variation of 33.4% and 40.5% for litters of clones and controls, respectively). Similarly there were no statistical differences between sex ratios of cloned litters (51-49%, M:F) and control litters (59-41%, M:F). Blood profiles between cloned pigs, control pigs, and their progeny were compared at two time points (i.e., 15 and 27 weeks) to quantify the effect of cloning on various blood parameters and their transmission to the next generation. Although the range of values for all traits overlapped between different classes, the variability between all the traits in F1 progeny of clones and the control pigs was similar at 15 and 27 weeks, with one exception. Combined, our data and previous results in mice strongly support the hypothesis that offspring of clones are similar to offspring of naturally bred animals, and as such there should not be any increased risks associated with consumption of products from these animals.     

Studies of the teratogenic potential of exposure of rats to 6000-MHz microwave radiation. II. Postnatal psychophysiologic evaluations

Wistar rats (36) were exposed daily throughout pregnancy to a power density level of 35 mW/cm2 of 6000-MHz microwave radiation (11), sham irradiated (10), or used as control animals (15). Litters were culled to a maximum of eight F1a offspring/litter (total = 124) on Postnatal Day 1 and subjected to a series of reflex tests beginning Day 3. Mothers were rebred 10 days after weaning. Teratologic evaluations were completed on 263 F1b offspring. Weekly weights were recorded for 298 F1a offspring. At 60 days, behavioral testing was initiated on 121 offspring. At 90 days, offspring were bred within/across groups. Teratologic evaluations were completed on 659 F2 term fetuses. Organ weight analyses were completed on 17 mothers and 181 F1a adult offspring, and blood analyses on 21 mothers and 131 offspring. Sex differences within groups were observed in four behavioral tests and in blood data. Significant differences between groups were observed for: F1b term fetal weight; F1a eye opening, postnatal growth to the fifth week, water T-maze and open field test results; and several organ/body weight ratios. These results indicate that exposure to 6000-MHz radiation at this power density level may result in subtle long-term neurophysiologic alterations not detectable at term using conventional morphologic teratologic procedures.     

Insemination of susceptible and preimmunized gilts with boar semen containing porcine reproductive and respiratory syndrome virus

Twenty-one gilts without measurable PRRSV serum antibody titres were identified for this experiment. Seven gilts were used as controls (Group C) and 14 as principals. Of these, 7 gilts were preimmunized to PRRSV and constituted Group B, while 7 gilts remained seronegative and constituted Group A. The principal gilts were inseminated with boar semen containing PRRSV and were killed 20 d later. The control gilts were treated similarly but were not exposed to PRRSV. Gilts were observed for clinical signs of infection. The effects on the conception rates were studied and gilts and embryos were tested for PRRSV and homologous antibodies. Group A and B gilts developed signs of PRRS associated with anorexia and slightly elevated body temperatures. Transmission of the infection was demonstrated by the isolation of PRRSV from serum and other tissue samples of principal gilts and also by seroconversion. The results show that early infection may have an insignificant effect or no effect on the conception and fertilization rates. However, exposure to PRRSV at the time of insemination can result in transplacental infection of embryos. In Group A gilts, 5 of 6 litters were infected prenatally with 7.6% of embryos infected. In Group B gilts, 1 of 5 litters and 1.3% of embryos were infected. Moreover, approximately 2 and 4 times more embryos were dead in litters of gilts from Group A and Group B than in gilts from control Group C. The isolation of PRRSV in 3 dead embryos suggests that the embryos may have died as a result of the direct effect of the virus. It can be concluded that the insemination of either seronegative or preimmunized gilts with boar semen containing PRRS V may have an insignificant effect or no effect on conception and fertilization rates, although it can result in transmission of the virus and embryonic infection and death.     

The effect of fetal hypophysectomy and fetal death in gilts with small litters on concentrations of relaxin,progesterone and estradiol-17β

During late pregnancy concentrations of relaxin, progesterone (P) and estradiol-17β (E) in maternal plasma were measured in gilts with small litters of intact, hypophysectomized, partially hypophysectomized or dead fetuses and in gilts with litters of normal fetuses and numbers. To achieve a small litter size at parturition all but one or two fetuses were killed at surgery at Day 30 to 40 of gestation. Fetal hypophysectomy or sham procedures were attempted on Day 90 to 95. Gestation was prolonged in gilts carrying hypophysectomized or partially hypophysectomized fetuses (P<0.01). Lactation and farrowing did not occur if hypophysectomy was complete. Basal concentrations of E in plasma were lower (P<0.01), basal P appeared higher and basal relaxin was unchanged in gilts carrying hypophysectomized or dead fetuses as compared to gilts with intact fetuses. Near the end of pregnancy the concentration of E was 119.8 pg/ml in gilts with the normal number of fetuses, 32.6 pg/ml in the group with hypophysectomized fetuses, and 7.3 pg/ml in gilts with dead fetuses. The relaxin peak occurred near term in control pigs and was delayed in the groups with hypophysectomized and partially hypophysectomized fetuses. The concentration of relaxin at the peak in gilts with normal sized litters was 181.4±75.8 ng/ml as compared with 25.3±16.0 ng/ml in gilts with partially hypophysectomized fetuses and was 9.5±1.4 ng/ml in gilts with hypophysectomized fetuses and 10.6±3.3 ng/ml in gilts with one or two fetuses. In gilts with intact or partially hypophysectomized fetuses, or litters containing both types, which came into labor, the patterns of P and E were similar. In gilts with hypophysectomized fetuses, P and E at term showed little change from basal concentrations. The results confirm that the fetus influences basal concentrations of E and possibly P in late normal gestations. In addition, the presence of the fetal pituitary is associated with the peak in relaxin expected at term. These associations are likely to be related to pituitary function and/or the mass of the conceptus. Fetal hypophysectomy is clearly associated with maternal concentrations of P and E at Day 114 that are different from those in normal sows, suggesting that these two hormones may have an effect on the initiation of parturition in the pig.     

Studies on prenatal and postnatal development in rats exposed to 60-Hz electric fields

A series of three experiments was performed to determine the effects of 30-day exposures to uniform 60-Hz electric fields (100 kV/m) on reproduction and on growth and development in the fetuses and offspring of rats. In the first experiment, exposure of females for 6 days prior to and during the mating period did not affect their reproductive performance, and continued exposure through 20 days of gestation (dg) did not affect the viability, size, or morphology of their fetuses. In the second experiment, exposure of the pregnant rat was begun on 0 dg and continued until the resulting offspring reached 8 days of age. In the third experiment, exposure began at 17 dg and continued through 25 days of postnatal life. In the second and third experiments, no statistically significant differences suggesting impairment of the growth or survival of exposed offspring were detected. In the second experiment, a significantly greater percentage of the exposed offspring showed movement, standing, and grooming at 14 days of age than among-sham-exposed offspring. There was a significant decrease at 14 days in the percentage of exposed offspring displaying the righting reflex in the second experiment and negative geotropism in the third experiment. These differences were all transient and were not found when the animals were tested again at 21 days of age. Evaluation of the reproductive integrity of the offspring of the second experiment did not disclose any deficits.     

Maternal transmission and dental caries induction in Sprague-Dawley rats infected with Streptococcus mutans

Thirty-four female rats (18 days old) were infected with Streptococcus mutans MT8148R (serotype c) or 6715 (g). Diets containing different proportions of sucrose were used to prepare the dams which harbored various levels of S. mutans in their oral cavity. Around 66 days of age, the female rats were bred and 34 dams subsequently bore 322 offspring. The dams were killed upon weaning (20 days of age) of their respective litters. There were positive correlations between the recovery of inoculated S. mutans and the caries incidence in the dams. Transmission of S. mutans from a dam to her offspring was studied in 10-, 15-, 20-, 27-, 34-, 41-, 48-, and 55-day-old rats by evaluating the recover of S. mutans from the offspring. Positive correlation between the magnitudes of recovered S. mutans MT8148R from dams and their offspring was found in all ages of young rats examined. Furthermore, caries incidence in young rats was found to be positively correlated with the recovery of both strains of S. mutans as well as with incidence of caries in their respective dams.     

Postovulatory aging of oocytes decreases reproductive fitness and longevity of offspring.

We analyzed the long-term effects of postovulatory aging of mouse oocytes on reproductive fitness and longevity of offspring. Hybrid (C57BL/6JIco x CBA/JIco) parental generation (F0) females were artificially inseminated at 13 h (approximately 1 h postovulation) or 22 h (approximately 10 h postovulation) after GnRH injection. Reproductive fitness of first generation (F1) females was tested from the age of 28 wk until the end of their reproductive life. In males, the testing period ranged from the age of 2 yr until their natural death. Experimental F1 females exhibited longer between-labor intervals, decreased frequency of litters, and lower total number of litters and offspring born. Experimental second generation (F2) pups displayed teratogenic defects, higher preweaning mortality, and decreased body weight at weaning. Incidence of infertility was higher in experimental F1 males, which translated into lower total number of offspring born when compared with the control group. Life expectancy of F1 offspring was decreased in the experimental group. These results clearly show that postovulatory aging of mouse oocytes decreases reproductive fitness and longevity of offspring.     

Repeat breeding and subsequent reproductive performance in Swedish Landrace and Swedish Yorkshire sows

The objective of the present study was to analyse the association between repeat breeding (RB) in gilts/sows and their subsequent reproductive performance as well as the impact of interactions between repeat breeding and factors like parity number, boar breed, season and mating type (MT) on subsequent reproductive performance in Swedish Landrace (L) and Swedish Yorkshire (Y) sows. Data analysed included 7040 sows (3654 L and 3386 Y), farrowing during January 1994 until December 1999 in 11 L and 8 Y nucleus herds. The study was assigned as a cohort design and the aim was to study gilts/sows from their first mating as gilts until mating after third parity. Analysis of variance was applied to continuous data and logistic regression was applied to categorical data. Percentages of litters as a result of repeat breeding in sow parities 1-3 were 6.1, 12.0 and 6.3% for L sows and 6.7, 13.1 and 7.4% for Y sows. For parity 3, the incidence of litters resulting from repeat breeding was significantly higher (P<0.001) in Y than in L sows. The proportion of irregular return to oestrus (>24 days after first mating) was higher (P<0.01) in primiparous sows than in multiparous sows (69% versus 61%). On average, litters resulting from repeat breeding were larger (P<0.001) than litters resulting from non-repeat breeding (NR) (about 0.5 piglets per litter) in both L and Y sows. For Y sows, if the previous litter was a result of repeat breeding, the subsequent reproductive cycle had 2.7% higher RR (P<0.05) and 2.4% lower FR (N.S.) compared with sows that were not repeat bred. The same trend was found in L sows (1.4% higher RR and 1.3% lower FR) but the differences were not significant. Among the sows removed from the herds, about 24% of L and 28% of Y were culled due to reproductive problems (gilts not included). In addition, a number of sows from these nucleus herds were also culled due to low breeding value and poor conformation.     

Pesticide Methoxychlor Promotes the Epigenetic Transgenerational Inheritance of Adult-Onset Disease through the Female Germline

Environmental compounds including fungicides, plastics, pesticides, dioxin and hydrocarbons can promote the epigenetic transgenerational inheritance of adult-onset disease in future generation progeny following ancestral exposure during the critical period of fetal gonadal sex determination. This study examined the actions of the pesticide methoxychlor to promote the epigenetic transgenerational inheritance of adult-onset disease and associated differential DNA methylation regions (i.e. epimutations) in sperm. Gestating F0 generation female rats were transiently exposed to methoxychlor during fetal gonadal development (gestation days 8 to 14) and then adult-onset disease was evaluated in adult F1 and F3 (great-grand offspring) generation progeny for control (vehicle exposed) and methoxychlor lineage offspring. There were increases in the incidence of kidney disease, ovary disease, and obesity in the methoxychlor lineage animals. In females and males the incidence of disease increased in both the F1 and the F3 generations and the incidence of multiple disease increased in the F3 generation. There was increased disease incidence in F4 generation reverse outcross (female) offspring indicating disease transmission was primarily transmitted through the female germline. Analysis of the F3 generation sperm epigenome of the methoxychlor lineage males identified differentially DNA methylated regions (DMR) termed epimutations in a genome-wide gene promoters analysis. These epimutations were found to be methoxychlor exposure specific in comparison with other exposure specific sperm epimutation signatures. Observations indicate that the pesticide methoxychlor has the potential to promote the epigenetic transgenerational inheritance of disease and the sperm epimutations appear to provide exposure specific epigenetic biomarkers for transgenerational disease and ancestral environmental exposures.     

Pre-weaning performance and health of pigs born to cloned (fetal cell derived) swine versus non-cloned swine

The objective of this study was to compare the pre-weaning performance of pigs derived from cloned versus non-cloned parents. Five cloned gilts and one cloned boar were used to produce five litters of pigs. One of five cloned females and the cloned boar were derived from two genetically unmanipulated fetal fibroblast cell lines. The remaining female clones were derived from a fetal fibroblast cell line in which random insertion of a alpha-1,3-galactosyltransferase gene targeting construct had occurred. Fetal cell lines had similar genetic backgrounds and were derived from three different fetuses in three different litters. Five litters of pigs were also generated from matings between two non-cloned boars and five non-cloned gilts. The mean gestation length, mean litter size, mean birth and weaning weights for male and female pigs were similar for litters derived from cloned parents versus non-cloned parents. The proportions of pigs born live and pigs that survived to weaning were also similar for pigs born to cloned as compared to non-cloned parents. In summary, matings between cloned swine derived from fetal fibroblast cell lines yielded litters of pigs that were similar in the number born, piglet birth weight and perinatal and pre-weaning mortality to litters produced by non-cloned swine.     

Effect of lactational exposure to tributyltin chloride on innate immunodefenses in the F1 generation in mice

We examined the effect of lactational exposure to tributyltin on innate immunodefenses in the F1 generation using in vivo and in vitro experiments. Pregnant C57BL/6 mice were given drinking water containing 0, 15, or 50 microg/ml of tributyltin chloride (TBTCl) from parturition to weaning. At weaning time, offspring were inoculated with Escherichia coli K-12, and bacterial clearances from the peritoneal cavity and spleen were examined. In vivo infection experiments indicated that bacterial clearance was significantly depressed in offspring breast-fed by dams exposed to 15 microg/ml of TBTCl (15 ppm F1), but not in offspring by dams exposed to 50 microg/ml of TBTCl (50 ppm F1). In vitro functional assays revealed that the killing activity of neutrophils decreased significantly in 15 ppm F1, but not in 50 ppm F1. We suggest that lactational exposure to TBT impairs innate immunodefenses in the F1 generation against non-pathogenic bacterial infection.     

Endocrine disrupting compounds: effect of octylphenol on reproduction over three generations

With the growing concern that environmental chemicals might impair human and animal fertility, it is important to investigate the possible influence of these substances on sexual differentiation and genital development of mammals. Many of these substances are suspected to interfere with endocrine processes, and exposure during critical periods of prenatal development might affect reproductive performance over several generations. Alkylphenols and their metabolites are lipophilic substances exerting apparent estrogenic action in in vitro and in vivo testing systems. With the widespread industrial use of alkylphenols, these are disseminated in the environment with sewage sludge, and domestic animals and humans are likely to be exposed via the food chain. Using the pig as an in vivo model, we studied the effect of intrauterine exposure to tertiary octylphenol (OP) on essential reproductive parameters over 3 generations. Sows were treated daily from D 23 to 85 of pregnancy with either 0, 10 or 1000 micrograms OP/kg body weight. Treatment with OP extended pregnancy length and induced basal cell proliferation in the cervical epithelium of the parental generation. In F1 offspring of sows treated with the low dosage of OP, onset of puberty was accelerated. Furthermore, when F1 gilts and F1 boars originating from sows treated with high dosages of OP were bred, the litter size was reduced. The results of the present study are compared with previous reports on estrogenicity of OP, and the usefulness of in vivo animal or embryo models for the evaluation of possible consequences of human exposure to endocrine disrupting compounds is discussed. Furthermore, possible consequences of exposure to endocrine disrupting compounds for the embryo transfer industry are addressed.     

Kin Recognition and Maternal Care under Restricted Feeding in House Mice (Mus domesticus)

This study tests the hypothesis that female house mice (F1 generation of wild caught Mus domesticus) should preferentially invest in own offspring if confronted with young of different degrees of relatedness. The maternal behaviour of females with litters of 4 own and 4 unrelated alien young (cross-fostered at day 1 of lactation) was analysed during a lactation period of 22 days both under ad libitum and under restricted feeding (food was restricted by 20%). Cross-fostering and restricted feeding had no effect on the amount of time spent nursing until weaning. Under both feeding conditions the females did not differ in their maternal behaviour towards own and alien young: there were no significant differences either in the amount of time spent nursing own versus alien pups or in the time spent licking own versus alien young. Weight gain of own and alien = wild littermates did not differ significantly in mixed litters and was similar both under ad libitum and under restricted feeding. Such indiscriminate behaviour might be adaptive if female house mice prefer to communally nest with a relative and thus improve their inclusive fitness by investing in own and related offspring in a communal nest. Under moderate restricted feeding females could not wean the entire litter but reduced litter size by cannibalizing on average 2.7 pups (75% of the pups were killed when they were 4–8 days old). Females with cross-fostered litters killed as many own as alien young. This suggests that females cannot discriminate between own and unrelated young if cross-fostering takes place at day 1 of lactation. Besides testing kin recognition abilities, the experiments also allow analysis of the weaning strategy of females under food shortage. Under restricted feeding, body weight of the females was significantly lower during middle lactation than under ad libitum feeding. Weaning weight of young in reduced litters under food restriction (9–10 g) did not differ significantly from weaning weight of young in litters of 7–10 young, but was lower than that of young in similar sized litters (litter size 6), under ad libitum feeding. The maternal behaviour of cannibalizing some young under food shortage can be interpreted as a weaning strategy which results in the largest number of offspring that can be raised to a minimal weaning weight of 9–10 g. Such a weaning strategy might represent a favourable trade-off between number and size of young produced.     

Long-term and transgenerational effects of cryopreservation on rabbit embryos

The short-term effects of cryopreservation and embryo transfer are well documented (reduced embryo viability, changes in pattern expression), but little is known about their long-term effects. We examined the possibility that embryo vitrification and transfer in rabbit could have an impact on the long-term reproductive physiology of the offspring and whether these phenotypes could be transferred to the progeny. Vitrified rabbit embryos were warmed and transferred to recipient females (F0). The offspring of the F0 generation were the F1 generation (cryopreserved animals). Females from F1 generation offspring were bred to F1 males to generate an F2 generation. In addition, two counterpart groups of noncryopreserved animals were bred and housed simultaneously to F1 and F2 generations (CF1 and CF2, respectively). The reproductive traits studied in all studied groups were litter size (LS), number born alive at birth (BA), and postnatal survival at Day 28 (number of weaned/number born alive expressed as percentage). The reproductive traits were analyzed using Bayesian methodology. Features of the estimated marginal posterior distributions of the differences between F1 and their counterparts (F1 − CF1) and between F2 and their counterparts (F2 − CF2) in reproductive characters found that vitrification and transfer procedures cause a consistent increase in LS and BA between F1 and CF1 females (more than 1.4 kits in LS and more than 1.3 BA) and also between F2 and CF2 females (0.96 kits in LS and 0.94 BA). We concluded that embryo cryopreservation and transfer procedures have long-term effects on derived female reproduction (F1 females) and transgenerational effects on female F1 offspring (F2 females).     

Allelic variation in the erythropoietin receptor gene is associated with uterine capacity and litter size in swine

A single nucleotide polymorphism (SNP; C vs. T) that creates an extra GATA-1 site (T allele) in intron 4 of the swine erythropoietin receptor (EPOR) gene was discovered and a genotyping assay for this SNP was developed. A total of 402 gilts from lines selected either at random (control), for ovulation rate (OR) or for uterine capacity (UC) for 11 generations were unilaterally hysterectomized-ovariectomized (UHO) at 160 days of age, mated at approximately 250 days of age and slaughtered at 105 days of pregnancy. Blood samples and spleens were collected from each foetus and the numbers of corpora lutea (CL) and live foetuses, the weights of each foetus and placenta, and each foetal haematocrit were recorded. In addition, intact gilts from the OR line or from a Yorkshire, Landrace, Duroc, crossbred line (BX) were mated and farrowed. At farrowing, the numbers of fully formed and live piglets were recorded for each litter. Genomic DNA was isolated for both the UHO and intact gilts, from foetuses from the UHO gilts that were heterozygous for the EPOR SNP, and from the boars from the BX line and were then used to determine EPOR SNP genotypes. Only CC and CT gilts were observed in the control, OR and UC selected lines. Presence of the EPOR T allele was associated (P < 0.05) with increased UC in these gilts. The number of heterozygous and homozygous foetuses did not differ within UHO litters, or did EPOR genotype influence foetal haematocrit. In intact gilts from the OR line, litter size was significantly associated (P < 0.05) with EPOR SNP genotype. Finally, results from intact gilts of the BX line, in which both the gilt and the boar genotypes were known, allowed an analysis to determine the effect of the gilt and/or the foetal genotype on litter size. This analysis indicated that the predicted foetal genotype (with gilt genotype as covariate) was associated with litter size (an increase of 2.6 +/- 1.0 piglets born alive predicted for homozygous T litters compared with homozygous C litters, P < 0.01) whereas the effect of the gilt genotype (adjusted for foetal genotype) on litter size was not significant. These results indicate that the EPOR SNP is associated with UC and litter size in two distinct populations and could be useful in increasing litter size in swine that are not limited in OR.     

Oestrone Sulphate Measurements for the Prediction of Small or Large Litters in Pigs

Serum from 88 pregnant sows and gilts was sampled 24 and 28 days after their first insemination or mating day. The oestrone sulphate (E1S) concentration in the samples was assessed with a commercially available radioimmunoassay kit modified for use with swine serum. The first aim was to test whether it was possible to predict litters of total number <10 piglets at term. The second aim was to compare the use of day 24 or day 28 samples, or of both, in this prediction.     

Maternal tobacco smoke exposure during lactation inhibits catecholamine production by adrenal medullae in adult rat offspring

Previously, we have shown that maternal smoke exposure during lactation, even when pups are not exposed, affects biochemical profiles in the offspring at weaning, eliciting lower body adiposity, hyperinsulinemia, hypocorticosteronemia and lower adrenal catecholamine content. However, the future impact of tobacco exposure is still unknown. As postnatal nicotine exposure causes short- and long-term effects on pups' biochemistry and endocrine profiles, we have now evaluated some endocrine and metabolic parameters of the adult offspring whose mothers were tobacco exposed during lactation. For this, from day 3 to 21 of lactation, rat dams were divided in: 1) SE group, cigarette smoke-exposed (1.7 mg nicotine/cigarettes for 1 h, 4 times/day, daily), without their pups, and 2) C group, exposed to air, in the same conditions. Offspring were killed at 180-days-old. Body weight and food intake were evaluated. Blood, white adipose tissue, adrenal, and liver were collected. All significant data were p<0.05. The adult SE offspring showed no change in body weight, cumulative food intake, serum hormone profile, serum lipid profile, or triglycerides content in liver. However, in adrenal gland, adult SE offspring showed lower catecholamine content ( - 50%) and lower tyrosine hydroxylase protein expression ( - 56%). Despite the hormonal alterations during lactation, tobacco smoke exposure through breast milk only programmed the adrenal medullary function at adulthood and this dysfunction can have consequence on stress response. Thus, an environment free of smoke during lactation period is essential to improve health outcomes in adult offspring.     

Placental insufficiency leads to developmental hypertension and mesenteric artery dysfunction in two generations of Sprague-Dawley rat offspring

It is generally accepted that preeclampsia results from reduction in perfusion to the uteroplacental unit leading to maternal hypertension and fetal growth restriction. Placental insufficiency creates an environment of fetal undernutriton, predisposing the fetus to the development of adult disease. In this study, we characterized the development and perpetuation of hypertension in two generations of male and female offspring subjected to an environment of fetal undernutrition via reduced uteroplacental perfusion pressure. Further, we examined vascular responses of resistance arteries in these animals to determine the influence of placental insufficiency on the development and perpetuation of hypertension. Experimental dams underwent a surgical procedure to reduce uteroplacental perfusion pressure, with resulting offspring comprising the first generation (F1). One male and one female from each of the F1 experimental litters served as breeders of the second generation (F2). Weekly systolic blood pressure measurements were obtained from 4 to 24 wk in control, F1, and F2 offspring. Vascular responsiveness to the vasoconstrictors phenylephrine and potassium chloride and the vasorelaxants acetylcholine and sodium nitroprusside was determined in the three offspring groups at 6, 9, and 12 wk of age. Our findings indicate that placental insufficiency during a critical developmental window in late gestation leads to hypertension in juvenile Sprague-Dawley rat offspring and is perpetuated in a second generation of offspring in a gender-specific manner. Further, exposure to placental insufficiency during late gestation leads to developmental alterations characterized by vascular hyperresponsiveness, perpetuated to a second generation of offspring in the absence of persistent environmental stimuli, contributing to hypertension.     

Increased postimplantation loss and malformations among the F2 progeny of male rats chronically treated with cyclophosphamide.

Cyclophosphamide, administered to the male rat, produces increased pre- and postimplantation loss in the progeny as well as an increase in the numbers of malformed and growth retarded fetuses. The purpose of this study was to determine whether the adverse effects of chronic paternal cyclophosphamide exposure are transmissible to the next generation, the F2 progeny. Adult male rats were treated by gavage daily with saline or with cyclophosphamide (3.4 or 5.1 mg/kg) for 4 or 18 weeks and mated. The male and female offspring in each treatment group (F1 generation) were randomly mated. The resulting pregnant females were killed on day 20 of gestation to evaluate progeny outcome in the F2 generation. There was a significant increase in postimplantation loss among the offspring of the group whose fathers had been treated with cyclophosphamide at a dose of 5.1 mg/kg/day. Exposure to a dose of 5.1 mg/kg/day of cyclophosphamide also resulted in an F2 generation with a significantly decreased mean fetal weight per litter and a significant increase in the number of malformed fetuses. The malformations observed among the F2 progeny included open eyes, omphalocele, generalized edema, syndactyly, gigantism, and dwarfism. Thus, exposure of the father to cyclophosphamide does result in a specific and heritable alteration in the fertility of the surviving "apparently normal" F1 progeny. Interestingly, the adverse consequences of exposure of male rats to cyclophosphamide are similar in the F2 generation to those previously reported for the F1 progeny.