Cheryl S. Rosenfeld

Maternal Diet and Other Factors Affecting Offspring Sex Ratio: A Review

Abstract Mammals usually produce approximately equal numbers of sons and daughters, but there are exceptions to this general rule, as has been observed in ruminant ungulate species, where the sex-allocation hypothesis of Trivers and Willard has provided a rational evolutionary underpinning to adaptive changes in sex ratio. Here, we review circumstances whereby ruminants and other mammalian species, especially rodents and primates, appear able to skew the sex ratio of their offspring. We also discuss some of the factors, both nutritional and nonnutritional, that potentially promote such skewing. Work from our laboratory, performed on mice, suggests that age of the mother and maternal diet, rather than the maternal body condition per se, play directive roles in controlling sex ratio. In particular, a diet high in saturated fats but low in carbohydrate leads to the birth of significantly more male than female offspring in mature laboratory mice, whereas when calories are supplied mainly in the form of carbohydrate rather than fat, daughters predominate. As the diets fed to the mice in these experiments were nutritionally complete and because litter sizes did not differ between treatments, dietary inadequacy seems not to be the cause for sex-ratio distortion. A number of mechanisms, all of which are testable, are discussed to provide an explanation for the phenomenon. We conclude the review by discussing potential implications of these observations to human medicine and agriculture.

Contrasting effects of different maternal diets on sexually dimorphic gene expression in the murine placenta

Diet during pregnancy influences the future health of a womans offspring, with outcomes differing depending on the childs sex. Because the placenta buffers the fetus from the mother, we examined the impact of diet and fetal sex on placental gene expression in mice fed either a very-high-fat, low-fat, chow diet of intermediate caloric density. At day 12.5 of pregnancy, placental RNA was extracted and analyzed by microarray. The expression of 1,972 genes was changed more than 2-fold (P < 0.05) in comparisons across diet in at least one of the three groups. Female placentae demonstrated more striking alterations in gene expression in response to maternal diet than male placentae. Notably, each diet provided a distinctive signature of sexually dimorphic genes, with expression generally higher in genes (651 out of 700) from female placentae than those from male placentae. Several genes normally considered as characteristic of kidney function were affected by diet, including genes regulating ion balance and chemoreception. The placenta also expressed most of the known olfactory receptor genes (Olfr), which may allow the placenta to sense odorant molecules and other minor dietary components, with transcript levels of many of these genes influenced by diet and fetal sex. In conclusion, gene expression in the murine placenta is adaptive and shaped by maternal diet. It also exhibits pronounced sexual dimorphism, with placentae of females more sensitive to nutritional perturbations than placentae of males.

Disruption of adult expression of sexually selected traits by developmental exposure to bisphenol A

Exposure to endocrine disrupting compounds (EDCs), such as bisphenol A (BPA), may cause adverse health effects in wildlife and humans, but controversy remains as to what traits are most sensitive to EDCs and might serve as barometers of exposure. Expression of sexually selected traits that have evolved through intrasexual competition for mates and intersexual choice of mating partner are more dependent on developmental and physical condition of an animal than naturally selected traits and thus might be particularly vulnerable to disruption by developmental exposure to EDCs. We have used the deer mouse (Peromyscus maniculatus) as a model to test this hypothesis. Adult male–male competition for mates in this species is supported by enhanced spatial navigational and exploratory abilities, which enable males to search for prospective, widely dispersed females. Male deer mice exposed to BPA or ethinyl estradiol (EE) through maternal diet showed no changes in external phenotype, sensory development, or adult circulating concentrations of testosterone and corticosterone, but spatial learning abilities and exploratory behaviors were severely compromised compared with control males. Because these traits are not sexually selected in females, BPA exposure predictably had no effect, although EE-exposed females demonstrated enhanced spatial navigational abilities. Both BPA-exposed and control females preferred control males to BPA-exposed males. Our demonstration that developmental exposure to BPA compromises cognitive abilities and behaviors essential for males to reproduce successfully has broad implications for other species, including our own. Thus, sexually selected traits might provide useful biomarkers to assess risk of environmental contamination in animal and human populations.

Transcription and Translation of Estrogen Receptor-β in the Male Reproductive Tract of Estrogen Receptor-α Knock-Out and Wild-Type Mice1

Estrogen receptor-α (ERα) has been identified in the male reproductive tract, but the role of estrogen in the male has not been well characterized. In vivo mutations in ERα genes have demonstrated the necessity for ERα-mediated action in male fertility. We asked whether both ERβ messenger RNA and protein were present in the male reproductive tract of wild-type and ERα knock-out (ERα KO) mice, and whether ERβ could compensate for the lack of ERα in infertile male ERα KO mice. Immunohistochemical localization with both N- and C-terminal anti-ERβ antibodies demonstrated that ERβ is present in the Leydig cells of the testes and in the epithelium of both the efferent ductules and the initial segment of the epididymis. RT-PCR amplification was used to confirm ERβ transcription in these tissues. In conclusion, we observed that ERβ messenger RNA and protein continue to be expressed in the Leydig cells, elongated spermatids, efferent ductules, and the initial segment of the epididymides of ERα KO mice, but the pre...

Striking variation in the sex ratio of pups born to mice according to whether maternal diet is high in fat or carbohydrate

In female mammals, it remains controversial whether maternal diet and particularly the source and availability of energy can influence sex of offspring born. Outbred female mice were fed ad libitum from 30 days to ≈45 wk of age on defined, complete diets that differed only in their relative content of fat and carbohydrate to determine whether calorie source influenced litter size and sex ratio of pups. Diet 1 (very high in saturated fat, VHF) provided 60% of calories as fat, mainly lard. Diet 2 (low in saturated fat, LF) was low in fat (10% of calories) but high in carbohydrate. Mice delivered four litters of pups, resulting in a total of 1,048 young born over 108 pregnancies. Gestation length and litter size did not differ between VHF and LF groups and did not change as mice aged. Sex ratio of pups (fraction male) born to mothers on VHF diet was unusually high (0.67) and to mothers on LF diet very low (0.39) over litters 2, 3, and 4. This skewing of sex ratio was related to diets fed and not to body mass of mothers. Age of mothers was an important variable, however. Mice that were first bred at 10 wk of age delivered similar numbers of sons and daughters, whereas virgin mice bred later than 20 wk of age produced litters that were skewed toward males or females according to diet. The data show that the source of calories provided in a nutritionally complete diet to mature female mice can influence sex of offspring born.

Effects of the environmental estrogenic contaminants bisphenol A and 17α-ethinyl estradiol on sexual development and adult behaviors in aquatic wildlife species.

Endocrine disrupting chemicals (EDCs), including the mass-produced component of plastics, bisphenol A (BPA) are widely prevalent in aquatic and terrestrial habitats. Many aquatic species, such as fish, amphibians, aquatic reptiles and mammals, are exposed daily to high concentrations of BPA and ethinyl estradiol (EE2), estrogen in birth control pills. In this review, we will predominantly focus on BPA and EE2, well-described estrogenic EDCs. First, the evidence that BPA and EE2 are detectable in almost all bodies of water will be discussed. We will consider how BPA affects sexual and neural development in these species, as these effects have been the best characterized across taxa. For instance, such chemicals have been in many cases reported to cause sex-reversal of males to females. Even if these chemicals do not overtly alter the gonadal sex, there are indications that several EDCs might demasculinize male-specific behaviors that are essential for attracting a mate. In so doing, these chemicals may reduce the likelihood that these males reproduce. If exposed males do reproduce, the concern is that they will then be passing on compromised genetic fitness to their offspring and transmitting potential transgenerational effects through their sperm epigenome. We will thus consider how diverse epigenetic changes might be a unifying mechanism of how BPA and EE2 disrupt several processes across species. Such changes might also serve as universal species diagnostic biomarkers of BPA and other EDCs exposure. Lastly, the evidence that estrogenic EDCs-induced effects in aquatic species might translate to humans will be considered.

Sex and dose-dependent effects of developmental exposure to bisphenol A on anxiety and spatial learning in deer mice (Peromyscus maniculatus bairdii) offspring.

Bisphenol A (BPA) is a widely produced, endocrine disrupting compound that is pervasive in the environment. Data suggest that developmental exposure to BPA during sexual differentiation of the brain leads to later behavioral consequences in offspring. Outbred deer mice (Peromyscus maniculatus bairdii) are an excellent animal model for such studies as they exhibit well-defined sex- and steroid-dependent behaviors. Here, dams during gestation and lactation were fed with a phytoestrogen-free control diet, the same diet supplemented with either ethinyl estradiol (0.1 ppb), or one of the three doses of BPA (50 mg, 5 mg, 50 μg/kg feed weight). After weaning, the pups were maintained on control diet until they reached sexual maturity and then assessed for both spatial learning capabilities and anxiety-like and exploratory behaviors. Relative to controls, males exposed to the two upper but not the lowest dose of BPA demonstrated similar impairments in spatial learning, increased anxiety and reduced exploratory behaviors as ethinyl estradiol-exposed males, while females exposed to ethinyl estradiol, but not to BPA, consistently exhibited masculinized spatial abilities. We also determined whether dams maintained chronically on the upper dose of BPA contained environmentally relevant concentrations of BPA in their blood. While serum concentrations of unconjugated BPA in controls were below the minimum level of detection, those from dams on the BPA diet were comparable (5.48±2.07 ng/ml) to concentrations that have been observed in humans. Together, these studies demonstrate that developmental exposure to environmentally relevant concentrations of BPA can disrupt adult behaviors in a dose- and sex-dependent manner.

Animal Models to Study Environmental Epigenetics

Abstract Epigenetics provides a means of understanding how environmental factors might alter heritable changes in gene expression without changing DNA sequence, and hence the origin, of some diseases that are not explained by conventional genetic mechanisms. Various animal models have been described, most notably the agouti viable yellow (Avy) and axin 1 fused (Axin1Fu) mice, which lend themselves particularly well to studying this link between epigenetics and development abnormalities, because particular changes in DNA methylation patterns can be linked to a broad spectrum of heritable pathologies in the mice. In addition, there are specific examples, both in mice and other animal species, where nonmutagenic, environmental insults to either parent, such as those caused by consumption of endocrine-disrupting chemicals, can cause unexpected transgenerational phenotypic changes in offspring. Animals derived by somatic cell nuclear transfer also frequently exhibit pathologies that can be linked to inappropriate nuclear reprogramming during early embryo development and provide another means to study a link between epigenetics and disease. This review will consider how all of these animal models might help elucidate the epigenetic basis for a wide range of phenotypes.

Sex-Specific Placental Responses in Fetal Development

The placenta is an ephemeral but critical organ for the survival of all eutherian mammals and marsupials. It is the primary messenger system between the mother and fetus, where communicational signals, nutrients, waste, gases, and extrinsic factors are exchanged. Although the placenta may buffer the fetus from various environmental insults, placental dysfunction might also contribute to detrimental developmental origins of adult health and disease effects. The placenta of one sex over the other might possess greater ability to respond and buffer against environmental insults. Given the potential role of the placenta in effecting the lifetime health of the offspring, it is not surprising that there has been a resurging interest in this organ, including the Human Placental Project launched by the National Institutes of Child Health and Human Development. In this review, we will compare embryological development of the laboratory mouse and human chorioallantoic placentae. Next, evidence that various species, including humans, exhibit normal sex-dependent structural and functional placental differences will be examined followed by how in utero environmental changes (nutritional state, stress, and exposure to environmental chemicals) might interact with fetal sex to affect this organ. Recent data also suggest that paternal state impacts placental function in a sex-dependent manner. The research to date linking placental maladaptive responses and later developmental origins of adult health and disease effects will be explored. Finally, we will focus on how sex chromosomes and epimutations may contribute to sex-dependent differences in placental function, the unanswered questions, and future directions that warrant further consideration.

Flawed Experimental Design Reveals the Need for Guidelines Requiring Appropriate Positive Controls in Endocrine Disruption Research

Frederick S. vom Saal,* Benson T. Akingbemi,† Scott M. Belcher,‡ David A. Crain,§ David Crews,{ Linda C. Guidice,jj Patricia A. Hunt,jjj Csaba Leranth,jjjj John Peterson Myers,# Angel Nadal,** Nicholas Olea,†† Vasantha Padmanabhan, Cheryl S. Rosenfeld, Alan Schneyer, Gilbert Schoenfelder, Carlos Sonnenschein, Ana M. Soto, Richard W. Stahlhut, Shanna H. Swan, Laura N. Vandenberg, Hong-Sheng Wang, Cheryl S. Watson, Wade V. Welshons, and Robert T. Zoeller