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

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

Open access
Wolstenholme, Jennifer T., Taylor, Julia A., Shetty, Savera R. J., Edwards, Michelle, Connelly, Jessica J., Rissman, Emilie F.·2011·PLoS ONE
DOI 10.1371/journal.pone.0025448PMC PMC3182223
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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.

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.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1371/journal.pone.0025448

Cites

74 of 74 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

30 of 77 cited sources checked · 18 supported · 1 flagged · accuracy not yet measured

  • Premisecitation

    A 40-fold higher BPA dose decreased male social preference in the identical task.

    “However a 40-fold higher BPA dose decreased male social preference in the identical task [16].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [16] K Cox, 2010, Gestational exposure to bisphenol A and cross-fostering affect behaviors in juvenile mice., Horm Behav source

      The Results (Social Novelty; page unavailable) report a diet-by-sex interaction in foster-reared juveniles, but identify increased investigation by BPA-exposed females rather than a decrease in male social preference. The Discussion (BPA Exposure Counteracts Sex Differences in Juvenile Social Behavior; page unavailable) describes BPA-exposed males as intermediate, not as showing a demonstrated decrease from control males. The Methods confirm a social-novelty task and a BPA dose, but the paper alone cannot establish the claimed 40-fold comparison. (LLM, conf=0.85)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
  • Premisecitation

    In humans, a positive association between gestational levels of BPA in mothers and externalizing behaviors in 2 year old girls has been reported.

    “In humans, a positive association between gestational levels of BPA in mothers and externalizing (hyperactivity and aggression) behaviors in 2 year old girls has been reported [19].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [19] JM Braun, 2009, Prenatal bisphenol A exposure and early childhood behavior., Environ Health Perspect source
  • Premisecitation

    In male mice exposed to low doses of BPA probably comparable to ours, mRNA for ERβ and several NMDA receptors were decreased in hippocampus at days 21 and 56.

    “In male mice, exposed to low doses of BPA that are probably comparable to ours, mRNA for ERβ and several of the NMDA receptors were decreased in hippocampus at day 21 and 56 [33].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [33] XH Xu, 2010, Perinatal exposure to bisphenol-A changes N-methyl-D-aspartate receptor expression in the hippocampus of male rat offspring., Environ Toxicol Chem source

      The Abstract and Results ('Expressions of NR subunits' and 'Expressions of ERb and P450arom'; page numbers unavailable) report reduced hippocampal NMDA-receptor subunit and ERβ expression after perinatal BPA exposure. But the offspring were male rats, not mice, and Western blots measured protein, not mRNA. The Results also report significant ERβ reduction at all postnatal time points for the highest dose, not for low doses at both days 21 and 56. (LLM, conf=0.93)

      Experimental — accuracy not yet measured
  • Study design and conductreasoning

    Pups were fostered to a control dam to limit offspring exposure to BPA 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].”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [16] K Cox, 2010, Gestational exposure to bisphenol A and cross-fostering affect behaviors in juvenile mice., Horm Behav source
    • cites [39] PL Palanza, 2002, Exposure to a low dose of bisphenol A during fetal life or in adulthood alters maternal behavior in mice., Environ Health Perspect source
    • Rests on an assumption: Fostering pups to a control dam successfully limits offspring BPA exposure to the gestational period.
  • Premisecitation

    Phytoestrogens in chow can enhance sex differences in the brain and in behavior.

    “Phytoestrogens in chow can enhance sex differences in the brain and in behavior [51], [52], [53], [54], [55].”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [51] RL Ruhlen, 2008, Low phytoestrogen levels in feed increase fetal serum estradiol resulting in the “fetal estrogenization syndrome” and obesity in CD-1 mice., Environ Health Perspect source

      The Objectives and Results compare reproductive and metabolic outcomes in male and female mice on different feeds, but do not report sex differences in the brain or behavior. Searches for brain, behavior, and sexual dimorphism found no passage supporting the attributed claim. (LLM, conf=0.72)

      Experimental — accuracy not yet measured
    • cites [52] R Ruhlen, 2011, Choice of animal feed can alter fetal steroid levels and mask developmental effects of endocrine disrupting chemicals., Journal of Developmental Origins of Health and Disease source
    • cites [53] KM Flynn, 2000, Effects of genistein exposure on sexually dimorphic behaviors in rats., Toxicol Sci source
  • May combine two claims. 31
  • Classified as probably not a substantial claim. 26

  • May combine two claims.

    Premisecitation

    Neonatal treatment with BPA reduces sex differences in the brain and can modify neurite and dendrite formation in rodents.

    “Neonatal treatment with BPA in general reduces sex differences in the brain and can modify neurite and dendrite formation [3], [8], [9], [10], [11].”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [3] HB Patisaul, 2006, Neonatal genistein or bisphenol-A exposure alters sexual differentiation of the AVPV., Neurotoxicol Teratol source
    • cites [8] T Hajszan, 2010, Bisphenol A interferes with synaptic remodeling., Front Neuroendocrinol source

      The section on testosterone-induced synaptogenesis (page unavailable) reports that BPA reduced spine synapses in adult male rats, supporting a narrower claim about altered brain structure. The perinatal-exposure section discusses developmental effects but says data on BPA’s interference with synapse development were lacking. The copy does not establish that neonatal BPA treatment generally reduces brain sex differences or modifies neurite and dendrite formation in rodents. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
    • cites [9] HB Patisaul, 2008, Influence of endocrine active compounds on the developing rodent brain., Brain Res Rev source

  • May combine two claims.

    Premisecitation

    BPA can influence social interactions and anxiety in rodents.

    “BPA can also influence social interactions and anxiety in rodents [7], [15], [16], [17].”
    Quote in the paperSubstantial claim1 of 3 sources checked
    • cites [7] BC Ryan, 2006, Developmental exposure to environmental estrogens alters anxiety and spatial memory in female mice., Horm Behav source
    • cites [15] F Dessi-Fulgheri, 2002, Effects of perinatal exposure to bisphenol A on play behavior of female and male juvenile rats., Environ Health Perspect source
    • cites [16] K Cox, 2010, Gestational exposure to bisphenol A and cross-fostering affect behaviors in juvenile mice., Horm Behav source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    BPA can act to alter DNA methylation.

    “In addition to steroid-related effects, BPA may have even more global actions as it can act to alter DNA methylation [29].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [29] DC Dolinoy, 2007, Maternal nutrient supplementation counteracts bisphenol A-induced DNA hypomethylation in early development., Proc Natl Acad Sci U S A source

  • Classified as probably not a substantial claim.

    Premisecitation

    Dysregulation of DNA methylation during critical developmental windows could disrupt the normal progression of brain and endocrine system development causing robust changes in the developing embryo that can persist into adulthood or even beyond if effects extend to germ cells.

    “Dysregulation of DNA methylation during these critical developmental windows could disrupt the normal progression of brain and endocrine system development causing robust changes in the developing embryo that can persist into adulthood or even beyond if effects extend to germ cells [30].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [30] JT Wolstenholme, 2011, The role of Bisphenol A in shaping the brain, epigenome and behavior., Horm Behav source

      Split into 2 atoms — 1 confirmed, 1 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Exposure to BPA is fairly ubiquitous, detected in urine in 90% of all humans sampled.

    “Exposure to this chemical is fairly ubiquitous; as it has been detected in urine in 90% of all humans sampled [1].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [1] AM Calafat, 2005, Urinary concentrations of bisphenol A and 4-nonylphenol in a human reference population., Environmental health perspectives source

      The Results report detectable BPA in 95% of 394 urine samples and describe exposure as widespread. The claim gives 90% instead and extends the finding to “all humans sampled,” whereas the Discussion says this sample was relatively small and not representative of the U.S. population. (LLM, conf=0.90)

      Experimental — accuracy not yet measured

  • Classified as probably not a substantial claim.

    Premisecitation

    Steroid hormones organize sex differences in the brain during neonatal development.

    “Steroid hormones organize sex differences in the brain during neonatal development [20], [21].”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [20] AP Arnold, 2009, The organizational-activational hypothesis as the foundation for a unified theory of sexual differentiation of all mammalian tissues., Hormones and behavior source
    • cites [21] P Negri-Cesi, 2004, Sexual differentiation of the brain: role of testosterone and its active metabolites., J Endocrinol Invest

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Study design and conductreasoning

    Because mRNA was assayed in whole brain and ERα expression is not global, BPA-induced changes in ERα expression may have been missed.

    “In neonatal cortex, estradiol hypermethylates ERα [61] and perhaps the same thing occurs here, but, given that we assayed mRNA in whole brain and ERα expression is not global we may have missed BPA induced changes in expression.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [61] ME Wilson, 2010, Estrogen receptor-alpha gene expression in the cortex: Sex differences during development and in adulthood., Horm Behav source

      The Introduction’s developmental-regulation section describes ERα expression in specific brain regions, supporting the premise that its expression is not uniform. It does not say that a whole-brain assay missed BPA-induced changes; that is the citing authors’ own possibility, while the citation in the printed sentence is attached to the earlier methylation clause. No page number is available in our copy. (LLM, conf=0.78)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    In that study, male C57BL/6J juvenile mice engaged in more social interactions, and females performed more play soliciting.

    “In that study, males engaged in more social interactions, and females performed more play soliciting.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [41] KH Cox, 2011, Sex differences in juvenile mouse social behavior are influenced by sex chromosomes and social context. source

  • Classified as probably not a substantial claim.

    Study design and conductcitation

    Pups were fostered to a control dam because differences in maternal behavior caused by BPA might affect behavior.

    “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].”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [16] K Cox, 2010, Gestational exposure to bisphenol A and cross-fostering affect behaviors in juvenile mice., Horm Behav source
    • cites [39] PL Palanza, 2002, Exposure to a low dose of bisphenol A during fetal life or in adulthood alters maternal behavior in mice., Environ Health Perspect source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Dnmt3a has been implicated in rewarding behavior and neuronal plasticity in the adult mouse nucleus accumbens.

    “Dnmt3a has been implicated in rewarding behavior and neuronal plasticity in the adult mouse accumbens [62].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [62] Q LaPlant, 2010, Dnmt3a regulates emotional behavior and spine plasticity in the nucleus accumbens., Nat Neurosci source

  • May combine two claims.

    Premisecitation

    Exposure to a 40-fold higher BPA dose decreased time in the middle, increased time in the closed arm, and tended to decrease time in the outermost portion of the open arm, indicating increased anxiety-related behaviors in the plus maze.

    “Yet again, exposure to the 40-fold higher BPA dose decreased time in the middle, increased time in the closed arm, and tended to decrease time in the outer most portion of the open arm [16], indicating that BPA at this high dose increased anxiety-related behaviors in the plus maze.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [16] K Cox, 2010, Gestational exposure to bisphenol A and cross-fostering affect behaviors in juvenile mice., Horm Behav source

      The Results (Elevated Plus Maze; page unavailable) report that BPA-exposed juveniles spent less time in the middle and more time in the closed arms. They also report less time in the distal open arm, particularly among males. The Methods and Results give the administered dose but do not establish the claimed 40-fold comparison; the plus-maze effects also varied by sex and rearing condition. (LLM, conf=0.86)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Slc1a1 is found throughout the cortex, hippocampus and basal ganglia and functions to buffer local glutamate at excitatory synaptic connections.

    “Slc1a1 is found throughout the cortex, hippocampus and basal ganglia [64] and functions to buffer local glutamate at excitatory synaptic connections [65].”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [64] JD Rothstein, 1995, Selective loss of glial glutamate transporter GLT-1 in amyotrophic lateral sclerosis., Ann Neurol source
    • cites [65] A Scimemi, 2009, Neuronal transporters regulate glutamate clearance, NMDA receptor activation, and synaptic plasticity in the hippocampus., J Neurosci source

  • May combine two claims.

    Premisecitation

    BPA readily crosses the placenta, exposing fetal tissues to concentrations close to those in maternal placenta.

    “BPA readily crosses the placenta [46], [47], exposing fetal tissues to concentrations close to those in maternal placenta [48].”
    Quote in the paperSubstantial claim1 of 3 sources checked
    • cites [46] B Balakrishnan, 2010, Transfer of bisphenol A across the human placenta., Am J Obstet Gynecol source
    • cites [47] M Nishikawa, 2010, Placental transfer of conjugated bisphenol A and subsequent reactivation in the rat fetus., Environ Health Perspect source

      The Results and Discussion (page unavailable) report that BPA-glucuronide crossed the placenta in pregnant rats and that small amounts of it and deconjugated BPA were detected in fetal tissues. They do not report fetal-tissue concentrations close to those in the maternal placenta; the Discussion instead describes the transfer as small. Thus, the source supports fetal exposure but not the concentration comparison attached to [48]. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [48] J Zhang, 2011, GC-MS analysis of bisphenol A in human placental and fetal liver samples., J Chromatogr B Analyt Technol Biomed Life Sci source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Studies directly assessing effects of fostering on maternal rearing behavior have conflicting results but raise the possibility that C57BL/6J dams may lick and/or groom fostered pups more than their biological pups.

    “The few studies directly assessing the effects of fostering on maternal rearing behavior have conflicting results [56], [57] but raise the possibility that C57BL/6J dams may lick and/or groom the fostered pups more than their biological pups.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [56] JP Curley, 2010, Developmental shifts in the behavioral phenotypes of inbred mice: the role of postnatal and juvenile social experiences., Behav Genet source
    • cites [57] R van der Veen, 2008, Impact of intra- and interstrain cross-fostering on mouse maternal care., Genes Brain Behav source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationcitation

    Genes on sex chromosomes may interact with BPA.

    “Another possibility is that genes on sex chromosomes may interact with BPA [58].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [58] AP Arnold, 2009, What does the “four core genotypes” mouse model tell us about sex differences in the brain and other tissues?, Front Neuroendocrinol source

      The Introduction discusses direct effects of X and Y genes, and section 7.2 discusses their possible effects on the brain and other tissues. Neither passage addresses an interaction with BPA; searches for “BPA” and “bisphenol” found no relevant passage in our copy. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The oxytocin receptor has long been associated with the display of social behavior in rodents.

    “The last candidate gene we examined, the oxytocin receptor, has long been associated with the display of social behavior in rodents [34].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [34] TR Insel, 2010, The challenge of translation in social neuroscience: a review of oxytocin, vasopressin, and affiliative behavior., Neuron source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    A polymorphic region of the human OXTR has been associated with empathy and stress reactivity.

    “A polymorphic region of the oxytocin receptor (OXTR) in humans has been associated with empathy and stress reactivity [73].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [73] SM Rodrigues, 2009, Oxytocin receptor genetic variation relates to empathy and stress reactivity in humans., Proc Natl Acad Sci U S A source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Two genome-wide association studies in autistic patients show linkage associations to chromosomal region 3p25.3, which contains OXTR.

    “Two genome wide association studies in autistics patients show linkage associations to chromosomal region 3p25.3, which contains OXTR [74], [75].”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [74] MB Lauritsen, 2006, A genome-wide search for alleles and haplotypes associated with autism and related pervasive developmental disorders on the Faroe Islands., Mol Psychiatry source
    • cites [75] JL McCauley, 2005, Genome-wide and Ordered-Subset linkage analyses provide support for autism loci on 17q and 19p with evidence of phenotypic and interlocus genetic correlates., BMC Med Genet source

      The Results report suggestive autism linkage at 3p25, and the Discussion says at least two other groups reported linkage at 3p. The paper describes a genome-wide linkage screen, not two genome-wide association studies; the cited passages do not establish the narrower 3p25.3 location or identify OXTR there (page numbers unavailable). (LLM, conf=0.88)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Findingown data

    No activity differences were detected as number of times mice crossed between three sections did not vary by group.

    “no activity differences were detected as the number of times that mice crossed between the three sections of the test box did not vary by group (Table 3).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    BPA exposure increased expression of Dnmt3a as main effects of diet were observed.

    “BPA exposure increased expression of Dnmt3a as main effects of diet were observed (F1,18 = 5.13, p<0.04).”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: The endogenous control genes B2M and β-actin are stably expressed across sex and BPA treatment.

  • May combine two claims.

    Findingown data

    Time spent in open arms, closed arms and number of crosses between arms were similar in all groups.

    “The time spent in the open arms, closed arms and the number of crosses between arms were similar in all groups (Table 4).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In the current study, there were no effects of sex or BPA on elevated plus maze behavior.

    “Likewise we did not find any effects of sex or BPA on elevated plus maze behavior in the current study.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    The largest gene expression effect was for Slc1a1, a glutamate transporter gene, whose expression was elevated in brains of female mice exposed to BPA.

    “Despite the lack of change in NMDA and AMPA receptors, the largest gene expression effect was noted for Slc1a1, one of the glutamate transporter genes, and interestingly its expression was elevated in brains of females exposed to BPA.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    BPA exposure altered Dnmt3a expression: the sex difference in controls (male > female) was absent in BPA-exposed brains because BPA increased Dnmt3a expression in females.

    “Dnmt3a, a gene responsible for de novo methylation, was altered by BPA exposure wherein the sex difference in control animals (male>females) was not present in BPA exposed brains because BPA increased Dnmt3a expression in females.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Using a dose well below the LOAEL, given only during gestation, produced several effects on juvenile social behaviors and embryonic whole-brain gene expression in mice.

    “Using a dose well below the LOAEL, given only during gestation, we detected several effects of BPA in diet on juvenile social behaviors as well as embryonic gene expression in whole brain, which may underlie the observed behavioral effects.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    DNA methyltransferases have been shown to have a number of interactions with estrogen receptors, particularly with ERβ.

    “In addition, these two mechanisms may act synergistically, as DNA methyltransferases have been shown to have a number of interactions with estrogen receptors, particularly with ER ß [31].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [31] S Pathak, 2010, Potential role of estrogen in regulation of the insulin-like growth factor2-H19 locus in the rat testis., Mol Cell Endocrinol source

  • May combine two claims.

    Interpretationown data

    Sex differences in juvenile social interactions are affected by BPA and in particular this compound modifies behavior in females.

    “Sex differences in juvenile social interactions are affected by BPA and in particular this compound modifies behavior in females.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Fostering pups to a control dam successfully limits offspring BPA exposure to the gestational period.

  • May combine two claims.

    Interpretationreasoning

    BPA exposure during gestation affects the brain and a number of behaviors in several mammalian species.

    “This convergence of data demonstrates that BPA exposure during gestation affects the brain and a number of behaviors in several mammalian species.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    BPA binds androgen and thyroid receptors.

    “It also binds the estrogen membrane receptor (GPER) with high affinity [23] as well as androgen and thyroid receptors [24], [25], [26], [27].”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [24] P Sohoni, 1998, Several environmental oestrogens are also anti-androgens., J Endocrinol source
    • cites [25] K Moriyama, 2002, Thyroid hormone action is disrupted by bisphenol A as an antagonist., The Journal of clinical endocrinology and metabolism source
    • cites [26] LC Xu, 2005, Evaluation of androgen receptor transcriptional activities of bisphenol A, octylphenol and nonylphenol in vitro., Toxicology source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    BPA is detectable in maternal plasma in ranges from 0.3 to 18.9 ng/ml and in fetal plasma from 0 to 9.2 ng/ml.

    “BPA is detectable in maternal and fetal plasma in ranges from 0.3 to 18.9 and 0 to 9.2 ng/ml respectively [2].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [2] G Schonfelder, 2002, Parent bisphenol A accumulation in the human maternal-fetal-placental unit., Environ Health Perspect source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    In cynomolgus monkeys, prenatal BPA exposure in male offspring is correlated with increased outward looking and exploration, and it affects behavior of the mothers.

    “In cynomolgus monkeys, prenatal BPA exposure in male offspring is correlated with increased outward looking and exploration, and it affects behavior of the mothers [18].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [18] A Nakagami, 2009, Alterations in male infant behaviors towards its mother by prenatal exposure to bisphenol A in cynomolgus monkeys (Macaca fascicularis) during early suckling period., Psychoneuroendocrinology source

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    Adult female mice were randomly assigned to control or BPA-supplemented diet.

    “Adult female C57BL/6J (B6) mice were randomly assigned to one of two groups and placed on either a phytoestrogen-free chow (n = 11; Harlan Teklad, TD95092) or the same chow supplemented with 1.25 mg BPA per kg diet (n = 12; Harlan Teklad, TD09710).”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Study design and conductown data

    Behavioral scoring was conducted by an observer blind to sex and treatment group.

    “Behaviors were recorded and later scored by an observer blind to sex and treatment group.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    BPA has steroid-like properties and binds to both estrogen receptors (ERα, ERβ) with low affinity.

    “BPA has steroid-like properties and binds to both estrogen receptors (ERα, ERβ) with low affinity [22], [23].”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [22] GG Kuiper, 1998, Interaction of estrogenic chemicals and phytoestrogens with estrogen receptor beta., Endocrinology source
    • cites [23] YB Wetherill, 2007, In vitro molecular mechanisms of bisphenol A action., Reprod Toxicol source

  • May combine two claims.

    Premisecitation

    Pre- and/or peri-natal exposure to BPA in rodents is associated with cognitive impairments and decreased exploration in a novel environment.

    “Pre- and/or peri-natal exposure to BPA in rodents is associated with cognitive impairments and decreased exploration in a novel environment [12], [13], [14].”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [12] YH Tian, 2010, Prenatal and postnatal exposure to bisphenol a induces anxiolytic behaviors and cognitive deficits in mice., Synapse source
    • cites [13] XH Xu, 2010, Perinatal exposure to bisphenol-A impairs learning-memory by concomitant down-regulation of N-methyl-D-aspartate receptors of hippocampus in male offspring mice., Hormones and behavior source
    • cites [14] A Poimenova, 2010, Corticosterone-regulated actions in the rat brain are affected by perinatal exposure to low dose of bisphenol A., Neuroscience source

  • May combine two claims.

    Premisecitation

    BPA exposure decreased the number of TH-positive cells in females, eliminating the sexual dimorphism in the AVPV of the hypothalamus.

    “For example, BPA exposure decreased the number of tyrosine hydroxylase (TH) positive cells in females, eliminating the sexual dimorphism in the anteroventral periventricular region of the hypothalamus [11].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [11] BS Rubin, 2006, Evidence of altered brain sexual differentiation in mice exposed perinatally to low, environmentally relevant levels of bisphenol A., Endocrinology source

  • May combine two claims.

    Premisecitation

    In a study of juvenile rat social behavior, BPA reduced social interest in both sexes, but a lower dose increased social approach and interactions in females.

    “In a study comparing male and female rat juvenile social behavior, BPA reduced social interest in both sexes but a lower dose exposure increased social approach and interactions in females [17].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [17] S Porrini, 2005, Early exposure to a low dose of bisphenol A affects socio-sexual behavior of juvenile female rats., Brain Res Bull source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Study design and conductreasoning

    Fostering at birth may have unidentified effects that alter the display of sex differences, and this cannot be excluded.

    “Second, mice in the present study were fostered at birth and we cannot exclude the possibility that fostering may have as yet unidentified effects which alter the display of sex differences.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Female mice exposed to BPA during brain sexual differentiation were less reactive and explored a novel environment less than control females.

    “Females exposed to BPA during the period of brain sexual differentiation were less reactive and explored a novel environment less than control females [49].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [49] L Gioiosa, 2007, Developmental exposure to low-dose estrogenic endocrine disruptors alters sex differences in exploration and emotional responses in mice., Horm Behav source

  • May combine two claims.

    Interpretationreasoning

    Endogenous estrogens may out-compete BPA for estrogen receptor binding sites in males but not in neonatal females lacking an endogenous hormone surge.

    “Thus endogenous estrogens may out-compete BPA, for example, for estrogen receptor binding sites in males but not in neonatal females that are not experiencing an endogenous hormone surge.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Study design and conductcitation

    Embryo use was limited to those positioned in utero next to at least one male littermate to reduce variation caused by intrauterine position.

    “Embryos were sexed by PCR [37] and we limited our use to embryos positioned in utero next to at least one male littermate to reduce any potential variation caused by intrauterine position [38].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [38] R Gandelman, 1977, Contiguity to male foetuses affects morphology and behaviour of female mice., Nature source

  • Probably not a substantial claim: may not pass S4 (based).

    Study design and conductown data

    Use of plastic in the assay was limited to pipet tips previously established not to leach BPA.

    “Use of plastic in the assay was limited to pipet tips (previously established not to leach BPA).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductown data

    Extraction efficiency averaged over 95%.

    “extraction efficiency was assessed from recovery of the internal standard, which averaged over 95%.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    BPA likely influenced ERα, because the oxytocin receptor is an ERα target gene and Oxtr mRNA was changed.

    “In fact it is likely that BPA did influence ERα since the oxytocin receptor is an ERα target gene and we noted changes in Oxtr mRNA [59], [60].”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [59] Y Kubota, 1996, Structure and expression of the mouse oxytocin receptor gene., Mol Cell Endocrinol source
    • cites [60] LJ Young, 1998, Estrogen receptor alpha is essential for induction of oxytocin receptor by estrogen., Neuroreport source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    In juvenile female rats, BPA increased both non-social and social investigation.

    “In juvenile females, BPA increased both non-social and social investigation [15].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [15] F Dessi-Fulgheri, 2002, Effects of perinatal exposure to bisphenol A on play behavior of female and male juvenile rats., Environ Health Perspect source

  • May combine two claims.

    Premisecitation

    Gestational BPA exposure disrupts amphetamine-induced conditioned place preference in female mice but not in male mice.

    “Amphetamine-induced conditioned place preference is also disrupted in female mice exposed to BPA during gestation while males displayed no change in place preference [50].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [50] G Laviola, 2005, D-amphetamine-related reinforcing effects are reduced in mice exposed prenatally to estrogenic endocrine disruptors., Brain Res Bull source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductown data

    The limit of detection for BPA in serum was 0.5 ng/ml.

    “The limit of detection (LOD) for BPA in serum by these methods was 0.5 ng/ml based on extraction of 0.5 ml of serum”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Slc1a1 null mice appear to have age-related impairments in self-grooming and spatial learning.

    “Behavioral characterization of the Slc1a1 null mouse is not extensive but with age these animals appear to have impairments in self-grooming and spatial learning [66].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [66] K Aoyama, 2006, Neuronal glutathione deficiency and age-dependent neurodegeneration in the EAAC1 deficient mouse., Nat Neurosci source
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

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • BPA alters AVP and OT neuron numbers and innervation of sexually dimorphic regions associated with social and aggressive behaviors in multiple species, and is anxiogenic in multiple species.

    “Altered AVP and OT neuron numbers and innervation of sexually dimorphic regions associated with social and aggressive behaviors in multiple species; anxiogenic in multiple species”

    From Endocrine Disruption of Vasopressin Systems and Related Behaviors (2017) · cites this paper as [64]

    The Results ('BPA exposure affects gene expression'; page unavailable) report altered oxytocin-receptor mRNA, not altered AVP or OT neuron numbers or innervation. The Discussion (page unavailable) reports that a 40-fold higher BPA dose increased anxiety-related behavior in mice, whereas the current low-dose study found no anxiety effect. The paper does not establish that BPA is anxiogenic in multiple species. (LLM, conf=0.87)

    Experimental — accuracy not yet measured

  • In C57BL/6J mice, gestational BPA exposure (dams on approximately 170 μg/kg bw diet) produced social deficits, particularly in females, coinciding with decreased AVP mRNA in whole embryonic brains of ED18.5 siblings of both sexes.

    “Mice reared on a diet delivering approximately 170 µg/kg bw BPA (to the dams) during gestation displayed social deficits, particularly in females, that coincided with a decrease in AVP mRNA expression levels in whole embryonic brains obtained from their ED18.5 siblings (both sexes).”

    From Endocrine Disruption of Vasopressin Systems and Related Behaviors (2017) · cites this paper as [64]

    The Methods (Animals) identify C57BL/6J mice and gestational exposure through BPA-supplemented chow, while the Discussion reports altered juvenile social behavior, especially in females. But the Discussion describes increased female social interactions, not social deficits. The Results (BPA exposure affects gene expression) report reduced oxytocin-receptor (Oxtr) mRNA in exposed male E18.5 brains; the paper does not report decreased AVP mRNA in both sexes. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • Reduced OT mRNA expression was observed in F4 male mice following BPA exposure.

    “Reduced OT mRNA expression was observed in the F4 males.”

    From Endocrine Disruption of Vasopressin Systems and Related Behaviors (2017) · cites this paper as [64]

    The Results section (“BPA exposure affects gene expression”; page unavailable) reports significantly reduced Oxtr mRNA in BPA-exposed male embryos. Oxtr encodes the oxytocin receptor, not oxytocin (OT), and the paper reports gestationally exposed embryos, not F4 males. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • BPA exposure causes female-specific alteration of one-on-one social interactions in juvenile mice and prairie voles.

    “as well as female-specific alteration of one-on-one social interactions in juvenile mice (Wolstenholme, 2011) and prairie voles (Sullivan et al., 2014).”

    From The effects of prenatal PCBs on adult social behavior in rats (2015) · cites this paper as Wolstenholme, 2011

  • In none of the three studies (Porrini et al., 2005; Dessi-Fulgheri et al., 2002; Wolstenholme et al., 2011) was the litter statistically controlled.

    “However, in none of those three studies was the litter statistically controlled.”

    From Developmental Treatment with Ethinyl Estradiol, but Not Bisphenol A, Causes Alterations in Sexually Dimorphic Behaviors in Male and Female Sprague Dawley Rats (2014) · cites this paper as Wolstenholme et al., 2011

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • BPA-female chamber times exceed the test duration

    Minor

    The BPA-female means for the three chambers sum to 602.3 seconds, exceeding the stated 10-minute (600-second) recording period. Rounding each mean to one decimal place cannot account for the excess.

  • Four genes are described as three

    Minor

    The parenthetical list names four genes, although the sentence calls them three.

  • Dose comparison uses the safe limit, not the LOAEL

    Minor

    For a 68 kg person, the stated LOAEL is 3,400 mg/day, which is 680,000 times the mice's stated 5 μg daily intake. The reported 680-fold comparison instead uses the 3.4 mg/day limit after the 1,000-fold safety factor.

  • Social-preference ANOVA degrees of freedom conflict with group sizes

    Minor

    The four Table 3 groups contain 67 mice. A two-way ANOVA with four groups and all 67 mice would have 63 residual degrees of freedom, not the reported 66.

4 of 4

Cites

Flagged first, then by citations

1 of the 75 references couldn’t be matched to a known paper.

Cited by

Sorted by citations

Related

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.
ORCID
Venue
PLoS ONE
Publisher
Public Library of Science (PLoS)
Published
2011-09-28
Volume
6
Issue
9
Pages
e25448
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0025448
PMC
PMC3182223
OpenAlex
W2013138247
Cited by
168
References
75
Citations by year
2026: 2 · 2025: 3 · 2024: 8 · 2023: 2 · 2022: 9 · 2021: 11 · 2020: 9 · 2019: 11 · 2018: 21 · 2017: 13 · 2016: 16 · 2015: 20 · 2014: 17 · 2013: 10 · 2012: 11
FWCI
8.16
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (submittedVersion, cc-by)
  • PubMed Central (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Topics
  • Physical Sciences › Environmental Science › Health, Toxicology and Mutagenesis › Effects and risks of endocrine disrupting chemicals
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › 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
MeSH terms
Bisphenol A Compounds, Animals, Animal Feed, Benzhydryl Compounds, Brain, Dose-Response Relationship, Drug, Female, Gene Expression Regulation

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-12.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-12.

Cited sources are checked by gpt-6-sol.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

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