Effects of levonorgestrel and STS 557 on testis in rodents

In immature male Wistar rats levonorgestrel and STS 557 (17 alpha-cyanomethyl-17 beta-hydroxyestra-4,9(10)-diene-3-one) reduced testicular growth, testicular DNA contents and plane of tubular cross-section in a dose dependent manner. In this respect, STS was less active than levonorgestrel. In immature castrated male rats the LH-suppressing effect of STS 557 was about 15 times lower than that of levonorgestrel. The results suggested that peripheral inhibitory effect of the two progestins was coupled with central LH suppression. On the other hand, in mating tests with male hybrid mice STS 557 possessed high fertility inhibiting activities in comparison to levonorgestrel and chlormadinone acetate. Limited dissociation has been shown between fertility inhibition and mating rate. All changes were reversible. Findings are discussed in view of development of male contraceptives.     

Tetrachlorodibenzo-p-dioxin exposure alters radial arm maze performance and hippocampal morphology in female AhR mice

Perinatal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been reported to alter spatial learning in rats tested on a radial arm maze (RAM). TCDD is believed to exert most of its effects through binding to the aryl hydrocarbon receptor (AhR). To determine whether the AhR mediates TCDD-induced alterations in spatial learning, we tested male and female AhR-knockout (AhR-/-), heterozygous (AhR+/-) and wild-type (AhR+/+) mice on the RAM. AhR+/- male and female mice were time mated, and treated dams were dosed with 5 microg TCDD/kg body weight on day 13 of gestation. When offspring reached adulthood, male and female AhR+/+, AhR+/- and AhR-/- mice from TCDD-exposed and unexposed litters were tested on the eight-arm RAM. After testing, we examined hippocampal morphology as visualized by the Timm's silver sulfide stain. TCDD-exposed female AhR+/- mice made more errors than their respective controls on the RAM and exhibited a decrease in the size of the intra- and infrapyramidal mossy fiber (IIP-MF) field of the hippocampus. None of the other TCDD-exposed groups differed from their respective control groups with regard to maze performance or hippocampal morphology. The reduction of IIP-MF field indicates a possible morphological basis for the learning deficit that was observed in the female AhR+/- mice. It is hypothesized that the effect of TCDD exposure is AhR dependent and that TCDD may alter GABAergic activity in the hippocampus of female mice during development.     

Prenatal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin produces alterations in cortical neuron development and a long-term dysfunction of glutamate transmission in rat cerebral cortex

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is considered one of the most toxic dioxin-like compounds. It is ubiquitous in foodstuffs of animal origin and accumulates in the fatty tissues of animals and humans. Prenatal TCDD exposure has been associated, beside other effects, with persistent impaired cognitive development. In the present study, the effects of maternal exposure to TCDD during pregnancy on cortical neuron development at birth and cortical glutamate transmission in new-born, 14- and 60-day-old rat offspring, were investigated. A single dose (0.7μg/kg) of TCDD dissolved in corn oil was orally administrated to the dams on gestational day 18; controls dams were treated with the vehicle. All the experiments have been performed on the male offspring from vehicle-treated (i.e. control group) and TCDD-treated dams. Primary cultures of cerebral cortical neurons obtained from 1-day-old rats born from mothers exposed to TCDD displayed a reduction in cell viability (MTT assay) and an increase in the number of apoptotic nuclei (nuclear staining with Hoechst 33258) possibly associated with altered dendrite outgrowth (MAP2-immunoreactivity) with respect to control cell cultures. These changes were associated with impairment in cortical glutamate transmission, characterized by a reduction in basal and K(+)-evoked outflow as well as a decrease in [(3)H]glutamate uptake. Interestingly, the prenatal TCDD-induced alteration of cortical glutamate signaling is persistent since it was also present in 14- and 60-day-old offspring. Taken together, these results suggest that a single prenatal exposure to TCDD produces alterations in cortical neuron development associated with a long-term dysfunction of glutamate transmission in rat cerebral cortex. The possible relevance of these findings for the understanding of the long-lasting cognitive deficit observed in the offspring from mothers exposed to the toxicant during pregnancy, is discussed.     

Teratogen update: lead and pregnancy

This review focuses on the impacts of lead exposure on reproductive health and outcomes. High levels of paternal lead exposure (>40 microg/dl or >25 microg/dl for a period of years) appear to reduce fertility and to increase the risks of spontaneous abortion and reduced fetal growth (preterm delivery, low birth weight). Maternal blood lead levels of approximately 10 microg/dl have been linked to increased risks of pregnancy hypertension, spontaneous abortion, and reduced offspring neurobehavioral development. Somewhat higher maternal lead levels have been linked to reduced fetal growth. Some studies suggest a link between increased parental lead exposure and congenital malformations, although considerable uncertainty remains regarding the specific malformations and the dose-response relationships. Common methodological weaknesses of studies include potential exposure misclassifications due to the frequent unavailability of exposure biomarker measurements at biologically appropriate times and uncertainty regarding the best exposure biomarker(s) for the various outcomes. A special concern with regard to the pregnant woman is the possibility that a fetus might be exposed to lead mobilized from bone stores as a result of pregnancy-related metabolic changes, making fetal lead exposure the result of exposure to exogenous lead during pregnancy and exposure to endogenous lead accumulated by the woman prior to pregnancy. By reducing bone resorption, increased calcium intake during the second half of pregnancy might reduce the mobilization of lead from bone compartments, even at low blood lead levels. Subgroups of women who incurred substantial exposures to lead prior to pregnancy should be considered to be at increased risk.     

Suppression of fetal testicular cytochrome P450 17 by maternal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin: a mechanism involving an initial effect on gonadotropin synthesis in the pituitary

The effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the fetal expression of testicular cytochrome P450 17 (CYP17), one of the enzymes necessary for sex steroid synthesis, was studied in Wistar rats. Fetal testicular CYP17 exhibited reduced mRNA and protein levels following exposure of the dams at gestational day 15 to 1 microg/kg TCDD. In support of this, CYP17 activity catalyzed by fetal testis homogenate was also reduced by maternal exposure to TCDD. The reduction in CYP17 expression seemed to be specific for fetal stages, because 7 day-old pups born from TCDD-treated dams did not exhibit any reduction in CYP17. In sharp contrast to the in vivo observations, TCDD failed to reduce CYP17 expression in cultured fetal testis, although CYP17 could be induced by activating cAMP-dependent signaling. To assess the role of pituitary luteinizing hormone (LH) on TCDD-induced reduction in fetal testicular CYP17, a further investigation was performed to examine whether the direct injection of LH into fetuses restores the altered CYP17 expression. The results showed that in utero injection of equine chorionic gonadotropin, an LH-mimicking hormone, completely abolishes the TCDD-produced reduction in fetal CYP17. However, neither the alpha- nor beta-subunits of LH in cultured fetal pituitary was reduced by TCDD. These results suggest that 1) maternal exposure to TCDD impairs the expression of testicular CYP17 in a fetal stage-specific manner; 2) this effect is due, at least partially, to a TCDD-produced reduction in circulating LH; and 3) TCDD exerts such an effect by affecting the upstream mechanism regulating the pituitary synthesis of LH.     

Lead exposure on critical days of fetal life affects fertility in the female mouse

Female mice were exposed to lead in utero by intravenous injection of lead chloride into the mothers at different stages of pregnancy. At a mature age the mice exposed as fetuses (F1 generation) conceived at a normal rate, but the litter size and fetal survival varied significantly. Small litters and increased numbers of fetal deaths were observed in mice exposed to lead on day 8 of intrauterine life. The live fetuses in this group were normal with respect to weight and morphological appearance. Serum levels of estradiol and progesterone, measured on day 17 of pregnancy, did not differ significantly between F1 mice of a control, unexposed group and of the group exposed to lead on day 8 of intrauterine life. Ovarian follicle counts revealed a significantly smaller number of primordial follicles in the latter group. It is suggested that the exposure to lead at a time of early organogenesis caused the observed fertility decrease by interfering with the development of the female germ cells.     

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.     

Effects of extremely low frequency magnetic field on fertility of adult male and female rats.

To investigate the effects of an extremely low-frequency (ELF) magnetic field on their fertility, adult male and female Sprague-Dawley rats were exposed to a 50 Hz sinusoidal magnetic field of approximately 25 microT (rms) for 90 days before they were mated with unexposed counterparts. Exposure to a 50 Hz field reduced male rat fertility. The number of pregnant females was reduced when mated with exposed males, and the number of resorptions increased. The effects of magnetic field on male fertility were shown to be partly reversible, when the same exposed group of males were remated 45 and 90 days after being removed from the fields. Exposure of adult female rats to 50 Hz magnetic fields for 90 days before mating significantly reduced their fertility. The mean numbers of implantations and living fetuses per litter were statistically significantly decreased in the 50 Hz group. These results suggest that low frequency magnetic fields have some adverse effects on fertility of male and female rats.     

Developmental and reproductive toxicology studies in IL-12p40 knockout mice

BACKGROUND: Interleukin (IL)‐12 is a cytokine that can exert regulatory effects on T and NK cells. This study was designed to identify potential developmental and reproductive hazards associated with IL‐12p40 knockout in mice. METHODS: In the combined fertility and teratology study, female F0 C57/BL6 wild‐type control mice and female F0 C57/BL6 IL‐12p40 homozgyous knockout mice were assessed for estrous cyclicity, sperm, and mating parameters. Pregnant females were euthanized on gestation day (GD) 18 and their fetuses were assessed for external, visceral, and skeletal development. In the peri and postnatal development study, the F1 wild‐type control and IL‐12p40 knockout mice were assessed for developmental landmarks, sexual development, passive avoidance, motor activity, and morris water maze. RESULTS: The IL‐12p40 knockout male mice exhibited decreased testis weights when compared to the wild‐type control group; however, this finding was not considered adverse, as it had no apparent functional effects on mating, fertility, and pregnancy rates or sperm motility. The IL‐12p40 knockout group exhibited effects on estrous cycle length, passive avoidance, morris water maze, and motor activity when compared to the wild‐type control group. However, since these findings were small in magnitude, transient and/or had no apparent effects on subsequent growth and development, they were not considered adverse. CONCLUSIONS: These results demonstrate that although IL‐12p40 homozygous knockout in mice exhibited effects on developmental and reproductive parameters, these effects were relatively minor and were not considered adverse. Birth Defects Res (Part B) 92:102–110, 2011. © 2011 Wiley‐Liss, Inc.     

Photoperiod influences reproduction in the prairie vole (Microtus ochrogaster)

Four experiments examined the role of photoperiod in the regulation of seasonal breeding in the prairie vole. Adult male voles maintained in short (8L:16D) as compared to long (16L:8D) photoperiods for 10 wk had reduced testicular and seminal vesicle weights, but fertility was not impaired. Male prairie voles reared from birth until 35 days of age in short as compared to long photoperiods also had reduced testicular and seminal vesicle weights, as well as diminished fertility. The incidence of pregnancy did not differ between long- and short-day female voles paired for 6 days with long- or short-day males (93%, 86%, 89%, and 88%, respectively). Photoperiod did not affect the incidence or the timing of postpartum pregnancies in long- or short-day females paired with long-day males through the birth of several litters. Adult male prairie voles retain only marginal sensitivity to short photoperiods, maturing males are highly responsive to short days, and adult females are insensitive to photoperiod. These data suggest that termination of the breeding season in the autumn may be due to the lack of fecund males in the population.     

Effects of sympathetic denervation on follicular distribution,oestradiol and progesterone serum levels in prepubertal hemi-ovariectomized female guinea pig

The aim of this study was to determine the effects of maternal undernutrition during pregnancy on adult reproductive function in male and female offspring. Groups of ewes were fed rations providing either 100% (High, H) or 50% (Low, L) of estimated metabolisable energy (ME) requirements for pregnancy, from mating until day 95 of gestation, and thereafter were conventionally managed. At 20 months of age, LH and FSH profiles, and LH responses to exogenous GnRH were measured in male and female offspring and, in males, testicular responses to exogenous LH (as measured by testosterone concentrations) were also measured. Undernutrition had no effect on the mean birth weights of lambs of either sex, or on testicular size in male animals at either 6 weeks or 20 months of age. L males exhibited significantly higher FSH concentrations than H males (P < 0.05) but there were no differences with treatment in FSH profiles in females, basal LH profiles or gonadotrophin responses to GnRH in offspring of either sex, and no difference in basal testosterone concentrations or in the testosterone response to exogenous LH administration in males. Semen quality at 20 months of age was unaffected by pre-natal undernutrition but ovulation rate was significantly reduced in L compared to H female offspring (P < 0.05). It is concluded that pre-natal undernutrition had no effect on male reproductive development and adult function, but reduced ovulation rate in female progeny. This effect was not associated with a change in gonadotrophin profiles or pituitary responsiveness.     

Host plant influence on the mating success of male Mediterranean fruit flies: variable effects within and between individual plants

Observations of field-caged guava trees, Psidium guajava, revealed that males of the Mediterranean fruit fly (medfly), Ceratitis capitata, occasionally formed tight aggregations on the trunk and branches of certain trees. Males at these aggregation sites (stations) appeared to feed on the bark, and chemical analyses showed that stations contained high levels of the male attractant α-copaene whereas ‘nonstations’ on the same tree lacked this chemical. Previous work showed that male medflies exposed to pure α-copaene had a mating advantage over unexposed males. Here, we present data showing that the occurrence of stations is highly variable both within and between individual guava trees. Concurrent mating trials showed that male medflies exposed to entire guava trees containing stations or to individual stations for 3 h gained a mating advantage over unexposed males in tests conducted 1 or 3 days following exposure. In contrast, males exposed to entire trees lacking stations or only to nonstations on trees with stations had similar mating success as unexposed males. Additional experiments showed that exposure to guava leaves had no effect on male mating frequency but that exposure to fruits enhanced male mating success. The discussion considers potential mechanisms underlying the plant-induced increase in mating success and potential effects of α-copaene and other plant-borne compounds on the spacing of medfly leks in the environment.     

Studies of the induction of dominant lethals and translocations in male mice after chronic exposure to microwave radiation

Male C3H mice were exposed to 100 W m-2 of 2.45 GHz continuous-wave microwave radiation for 6 h per day for a total of 120 h over an 8-week period. The exposure level was chosen so that the specific energy absorption rate (SAR) would be approximately equal to the level of 4 W kg-1 which is considered by a number of organizations to be a threshold for adverse biological effects. At the end of the treatment period the mice were mated with a different group of (C3H x 101) F1 hybrid females each week for the following 8 weeks. There was no significant reduction in pregnancy rate, preimplantation survival or postimplantation survival in the exposed group compared to sham-exposed controls. At the end of the mating period a cytogenetic analysis was carried out of meiotic chromosome preparations of testicular tissue, thus sampling cells that were stem cell spermatogonia during the treatment regime. The results showed no difference in the frequency of reciprocal translocations between the sham and treated groups, or in the frequency of cells with autosome or sex chromosome univalents. Low levels of fragments and exchanges were found in both groups. It is concluded that there is no evidence in this experiment to show that chronic exposure of male mice to 2.45 GHz microwave radiation induces a mutagenic response in male germ cells. This conclusion is in agreement with the observations of Berman et al. (1980), who reported a lack of male germ cell mutagenesis after repetitive or chronic exposure of rats to 2.45 GHz.     

Time Course for Onset and Recovery from Effects of a Novel Male Reproductive Toxicant: Implications for Apical Preclinical Study Designs

In the pharmaceutic ICH S5(R2) guidelines for reproductive toxicity testing, a premating dose duration of 14 days is considered sufficient for assessment of male fertility for compounds that are not testicular toxicants. A novel α7 subtype of nicotinic acetylcholine receptor (α7nAChR) agonist, originally intended for treatment of Alzheimer's disease, did not cause changes in sperm counts, motility, or testicular histopathology in rat toxicity studies of up to 6 months duration. However, profound decrements in male fertility (reduced pregnancy rates and litter sizes) occurred after 11 weeks of dosing in male rats. In two time‐course investigations, dosed male rats were paired with undosed females after 5, 14, and 28 daily doses and again after 2 and 4 weeks off‐dose. Effects on male fertility were undetectable after 5 days. After 14 days, there was no effect on pregnancy rate, but preimplantation losses were increased. Effects on both pregnancy rates and preimplantation losses were clearly detectable after 28 days, but were of lesser magnitude than after 11 weeks of dosing. Fertility recovered rapidly after dose cessation. These studies illustrate the sensitivity of a long premating dose period at revealing hazard and determining the magnitude of effect on male fertility for compounds that are intended for chronic administration and do not affect testicular histopathology     

Polyandry produces sexy sons at the cost of daughters in red flour beetles

Female mating with multiple males within a single fertile period is a common phenomenon in the animal kingdom. Female insects are particularly promiscuous. It is not clear why females mate with multiple partners despite several potential costs, such as expenditure of time and energy, reduced lifespan, risk of predation and contracting sexually transmitted diseases. Female red flour beetles (Tribolium castaneum) obtain sufficient sperm from a single insemination to retain fertility for several months. Nonetheless they copulate repeatedly within minutes with different males despite no direct fitness benefits from this behaviour. One hypothesis is that females mate with multiple partners to provide indirect benefits via enhanced offspring fitness. To test this hypothesis, we compared the relative fitness of F(1) offspring from females mated with single males and multiple males (2, 4, 8, or 16 partners), under the condition of relatively high intraspecific competition. We found that a female mating with 16 males enhanced the relative fitness of F(1) males (in two out of three trials) but reduced F(1) females' fitness (in two independent trials) in comparison with singly mated females. We also determined whether several important fitness correlates were affected by polyandry. We found that F(1) males from mothers with 16 partners inseminated more females than F(1) males from mothers with a single partner. The viability of the eggs sired or produced by F(1) males and females from highly polyandrous mothers was also increased under conditions of low intra-specific competition. Thus, the effects of polyandry on F(1) offspring fitness depend on environmental conditions. Our results demonstrated a fitness trade-off between male and female offspring from polyandrous mothers in a competitive environment. The mechanisms and biological significance of this unique phenomenon are discussed.     

Long-term exposure of male and female mice to 50 Hz magnetic field: effects on fertility

The effect of an extremely low frequency (ELF) magnetic field on the fertility of adult male and female Swiss mice was investigated. Adult male and female mice were exposed to a 50 Hz sinusoidal magnetic field at approximately 25 microT (rms) for 90 days before they were mated with unexposed counterparts. There were no exposure related effects on the fertility of male or female mice. The number of implantation sites, viable fetuses, and the total number of resorptions were not significantly affected in females impregnated by males exposed to the 50 Hz magnetic field as compared with the control group. The number of implantation sites, viable fetuses and the total number of resorptions in exposed females were also not statistically different from the control group. There were no significant effects on the weights of the testes, seminal vesicles, preputial gland or body weights of males exposed to 50 Hz magnetic field. Furthermore, body and uterine weights were not affected in females exposed to 50 Hz field; however, ovarian weight was significantly increased in females exposed to the same field. These results suggest that exposure of male and female mice to low frequency magnetic field had no adverse effects on fertility and reproduction in mice.