Cited by 169Effects and risks of endocrine disrupting chemicalsSexual Differentiation and DisordersEpigenetics and DNA Methylation

The DOI matches its Crossref record; 74 of the 75 references it cites were found in a registry or an index; 30 of the 77 sources its claims cite were checked for support.

Gestational Exposure to Low Dose Bisphenol A Alters Social Behavior in Juvenile Mice

Open access

Abstract

Bisphenol A (BPA) is a man-made compound used to make polycarbonate plastics and epoxy resins; public health concerns have been fueled by findings that BPA exposure can reduce sex differences in brain and some behaviors. We asked if a low BPA dose, within the range measured in humans, ingested during pregnancy, would affect social behaviors in prepubertal mice. We noted sex differences in social interactions whereby females spent more time sitting side-by-side, while males engaged in more exploring and sitting alone. In addition BPA increased display of nose-to-nose contacts, play solicitations and approaches in both sexes. Interactions between sex and diet were found for self grooming, social interactions while sitting side-by-side and following the other mouse. In all these cases interactions were produced by differences between control and BPA females. We examined brains from embryos during late gestation to determine if gene expression differences might be correlated with some of the sexually dimorphic or BPA affected behaviors we observed. Because BPA treatments ended at birth we took the brains during embryogenesis to increase the probability of discovering BPA mediated effects. We also selected this embryonic age (E18.5) because it coincides with the onset of sexual differentiation of the brain. Interestingly, mRNA for the glutamate transporter, Slc1a1, was enhanced by exposure to BPA in female brains. Also we noted that BPA changed the expression of two of the three DNA methyltransferase genes, Dnmt1 and Dnmt3a. We propose that BPA affects DNA methylation of Sc1a1 during neural development. Sex differences in juvenile social interactions are affected by BPA and in particular this compound modifies behavior in females.

Overview

AI snapshot

Written from the paper’s full text.

TL;DR

Gestational low-dose BPA exposure in mice altered juvenile social behavior, especially increasing social interactions in females, and changed embryonic brain expression of Slc1a1 and Dnmt genes. At this dose, BPA did not affect juvenile social preference or anxiety-like behavior.

Key findings

  • Sex differences were present: males spent more time exploring the cage and sitting alone, while females spent more time sitting side-by-side (F1,63 = 6.8, 4.91, and 3.92, respectively; p<0.05 at least).
  • BPA increased nose-to-nose contacts and approaches in both sexes (F1,63 = 3.99 and 3.82; p<0.05). Diet × sex interactions occurred for side-by-side non-grooming interactions (F1,63 = 8.13, p<0.006), following (F1,63 = 5.01, p<0.03), and self-grooming (F1,63 = 5.51, p<0.02); all were driven by BPA versus control females, with BPA females more interactive and self-grooming less.
  • At embryonic day 18.5, Slc1a1 was sexually dimorphic (F1,18 = 4.58, p<0.05) and showed a diet × sex interaction (F1,18 = 5.84, p<0.04), with BPA females highest. Dnmt3a was increased by BPA (F1,18 = 5.13, p<0.04) and sexually dimorphic (F1,18 = 5.49, p<0.04; males > females). Dnmt1 showed a trend interaction (F1,18 = 3.68, p<0.07), lowest in BPA females, and Oxtr showed a trend interaction (F1,18 = 4.14, p = 0.06), with BPA males lower.
  • BPA did not alter social preference or elevated plus maze behavior; social preference was sexually dimorphic (F1,66 = 4.43, p<0.05). Serum BPA was 0.099±0.014 ng/ml in control dams (n = 3) and 0.43±0.002 ng/ml in BPA-fed dams (n = 2), within the reported human maternal range of 0.3–18.9 ng/ml.

Methods

  • Pregnant C57BL/6J dams received phytoestrogen-free chow (n = 11) or chow supplemented with 1.25 mg BPA/kg diet (n = 12). Embryonic brains were collected at E18.5 from 10 litters (BPA n = 5, control n = 5), sexed by PCR, and restricted to embryos positioned next to at least one male littermate. qPCR measured Esr1, Esr2, Gper, Oxtr, Dnmt1/3a/3b, Gria1, Grin2a/b, and Slc1a1, normalized to B2M or β-actin using delta-delta Ct.
  • Behavior: pups were fostered at birth to control dams; BPA offspring n = 39 (18 females, 21 males), control n = 28 (13 females, 15 males). On PN21, mice underwent a 30-minute same-sex, same-age, same-treatment dyadic social interaction test; PN22 elevated plus maze; PN24 three-chamber social preference test with an adult male stimulus. Scoring was blind to sex and treatment.
  • Statistics: two-way ANOVA with sex and diet as factors, followed by Fisher post hoc tests with corrections for multiple comparisons; BPA serum was measured by HPLC.

Limitations

  • Stated: fostering introduces new issues, though all animals were fostered to control dams to control for maternal care differences; the authors cannot exclude fostering effects on sex differences. Whole-brain qPCR may miss regional BPA-induced changes, such as in ERα. Treatment ended at birth, and embryonic gene expression was sampled at E18.5.
  • Evident: only one BPA dose was tested, so no dose-response can be assessed. The BPA serum assay had very small sample sizes (control n = 3; BPA n = 2 in the results), and gene expression used n = 5–6 per group.
  • Evident: DNA methylation was not directly measured; the proposed methylation mechanism is based on Dnmt mRNA changes and is speculative.

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Integrity signals

Registry

Crossref · 10.1371/journal.pone.0025448

Cites

74 of 74 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Figures

Figure 1 Mean +/- SEM behaviors displayed during a reciprocal social interaction task. Juvenile male and female mice were exposed to control (phytoestrogen-free chow, black histograms, n = 15 males and n = 13 females) or BPA supplemented chow (gray histograms, n = 21 males and n = 18 females) during gestation. A) Total amount of time in seconds spent exploring the cage. B) Duration of side by side sitting (seconds). C) Amount of time spent self grooming (seconds). D) Duration of side by side interactions (seconds). * Sex effect; males are significantly different from females, p<0.05. # Significantly different from control females, p<0.05.
Figure 1 Mean +/- SEM behaviors displayed during a reciprocal social interaction task. Juvenile male and female mice were exposed to control (phytoestrogen-free chow, black histograms, n = 15 males and n = 13 females) or BPA supplemented chow (gray histograms, n = 21 males and n = 18 females) during gestation. A) Total amount of time in seconds spent exploring the cage. B) Duration of side by side sitting (seconds). C) Amount of time spent self grooming (seconds). D) Duration of side by side interactions (seconds). * Sex effect; males are significantly different from females, p<0.05. # Significantly different from control females, p<0.05.CC BY · PubMed Central
Figure 2 Mean +/- SEM mRNA expression measured with qPCR in embryonic day 18.5 brain relative to control males and normalized to beta-2 microglobulin or beta-actin (n = 5–6/group). Black bars indicate groups exposed to control, phytoestrogen-free diet. Gray bars indicate groups exposed to BPA during gestation. A) Oxytocin receptor (Oxtr). B) Neuronal glutamate transporter (Scl1a1). C) DNA methlytransferase 1 (Dnmt1). D) DNA methyltransferase 3a (Dnmt3a). *Significantly different from all other groups, p<0.05.
Figure 2 Mean +/- SEM mRNA expression measured with qPCR in embryonic day 18.5 brain relative to control males and normalized to beta-2 microglobulin or beta-actin (n = 5–6/group). Black bars indicate groups exposed to control, phytoestrogen-free diet. Gray bars indicate groups exposed to BPA during gestation. A) Oxytocin receptor (Oxtr). B) Neuronal glutamate transporter (Scl1a1). C) DNA methlytransferase 1 (Dnmt1). D) DNA methyltransferase 3a (Dnmt3a). *Significantly different from all other groups, p<0.05.CC BY · PubMed Central

Verification

Claims the paper makes

52 Own result · 22 No full text of the sources · 12 Supported · 9 Partly supported · 9 Own method · 8 Own interpretation · 3 Supported so far · 2 Couldn’t confirm · 1 Not supported · 1 Not checked yet · 1 No basis given · accuracy not yet measured

Show all 72 claims

  • It may state two separate claims as one. 31
  • Probably not a substantial claim. 26
48 of 48
  • The paper relies on this without support.

    Fostering pups to a control dam successfully limits offspring BPA exposure to the gestational period.

    “We did this to limit offspring exposure to BPA to the gestational period and because differences in maternal behavior caused by BPA might affect behavior [16], [39].”
  • The paper relies on this without support.

    The endogenous control genes B2M and β-actin are stably expressed across sex and BPA treatment.

    “normalized to B2M for TaqMan assays and ß-actin for SYBR Green assays as endogenous controls.”
  • The authors state this as an assumption.

    BPA levels achieved in mouse dams were also achieved in embryos.

    “Incorporating BPA in chow we produced tonic levels in mouse dams comparable to humans and we assume these were also achieved in the embryos.”
3 of 3

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Metadata

Authors
Wolstenholme, J. T., Taylor, J. A., Shetty, S. R. J., Edwards, M., Connelly, J. J., & Rissman, E. F.
Affiliations
  • University of Virginia (US)
  • University of Missouri (US)
Corresponding author
Rissman, Emilie F.
Venue
PLoS ONE
Published
2011-09-28
Volume
6
Issue
9
Pages
e25448
ISSN
1932-6203 (ISSN-L)
OpenAlex
W2013138247
Cited by
169
References
75
Citations by year
2026: 3 · 2025: 3 · 2024: 8 · 2023: 2 · 2022: 10 · 2021: 11 · 2020: 9 · 2019: 12 · 2018: 22 · 2017: 14 · 2016: 15 · 2015: 21 · 2014: 17 · 2013: 10 · 2012: 12
FWCI
8.16
Type
journal article
Language
en
Open access
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Autism Speaks (AS4802)
  • National Institute of Mental Health (R01MH086711)
  • National Institute of Diabetes and Digestive and Kidney Diseases (T32DK007646)
  • National Institute of Environmental Health Sciences (F32ES01940, F32ES019404)
Topics
  • Physical Sciences › Environmental Science › Health, Toxicology and Mutagenesis › Effects and risks of endocrine disrupting chemicals
  • Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Sexual Differentiation and Disorders
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
Keywords
bisphenol A, prenatal exposure, social behavior, sex differences, juvenile mice, social interaction, DNA methylation, DNMT1, DNMT3A, SLC1A1, glutamate transporters, embryonic brain development, sexual differentiation of the brain, self-grooming
MeSH terms
Bisphenol A Compounds, Animals, Animal Feed, Benzhydryl Compounds, Brain, Dose-Response Relationship, Drug, Female, Gene Expression Regulation, Humans, Male, Phenols, Pregnancy, Prenatal Exposure Delayed Effects, Receptors, Estrogen, RNA, Messenger, Sex Characteristics, Social Behavior, Receptors, Oxytocin, Mice

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