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Cited by 33Neuroendocrine regulation and behaviorSexual Differentiation and DisordersHypothalamic control of reproductive hormones

Unexpected Effects of Perinatal Gonadal Hormone Manipulations on Sexual Differentiation of the Extrahypothalamic Arginine-Vasopressin System in Prairie Voles

Open access
Lonstein, Joseph S., Rood, Benjamin D., De Vries, Geert J.·2005·Endocrinology
DOI 10.1210/en.2004-1315PMC PMC1444947
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Abstract

The sexually dimorphic extrahypothalamic arginine-vasopressin (AVP) projections from the bed nucleus of the stria terminalis to the lateral septum (LS) and lateral habenula (LHb) are denser in males than females and, in rats, require males' perinatal exposure to gonadal hormones but the absence of such exposure in females. We examined perinatal hormone effects on development of this sex difference in prairie voles (Microtus ochrogaster), which show atypical effects of hormones on sexual differentiation of some reproductive behaviors. Neonatal castration reduced the number of AVP mRNA-expressing cells in the bed nucleus of the stria terminalis and AVP immunoreactivity (ir) in the LS and LHb. Surprisingly, daily injections of 1000 microg of testosterone propionate (TP) during the first postnatal week did not maintain high levels of AVP-ir in neonatally castrated males. Furthermore, perinatal treatments with TP (75, 500, or 1000 microg), testosterone (100 microg), or dihydrotestosterone (200 microg) did not masculinize AVP-ir in the female LS or LHb. In fact, 1000 microg TP reduced it in some cases. However, 1000 microg TP lengthened anogenital distance, indicating that TP was biologically active. Neonatal estrogen receptor antagonism with tamoxifen reduced AVP-ir in the male LS, whereas treating neonatal females with the synthetic estrogen diethylstilbestrol increased septal AVP-ir. Tamoxifen and diethylstilbestrol had no effects in the LHb. Similar to rats, therefore, postnatal estrogen influences some components of the extrahypothalamic AVP system in prairie voles, but this developing system appears to be insensitive to exogenous androgens, including aromatizable androgens. Such insensitivity is atypical for a sexually dimorphic neural system in a rodent and may reflect the unusual effects of hormones on sexual differentiation of some behaviors in prairie voles.

AI snapshot

Written from the paper’s full text.

TL;DR

In prairie voles, neonatal castration demasculinized extrahypothalamic AVP expression, but perinatal treatment with exogenous androgens (TP, T, DHT) failed to maintain or masculinize AVP-ir in males or females, whereas estrogenic manipulations (tamoxifen in males, DES in females) altered septal AVP. These results suggest that postnatal estrogen influences some components of the extrahypothalamic AVP system in prairie voles, but this developing system is insensitive to exogenous androgens, unlike rats.

Key findings

  • Neonatal castration reduced AVP mRNA-expressing cells in the BST and AVP-ir in the LS and LHb. In Exp 1, control males had more LS AVP-ir than control females, with castrated males intermediate (F(2,27)=9.19, P≤0.001); LHb showed a similar pattern (F(2,28)=7.71, P≤0.003). In Exp 2, sham-castrated males had the greatest number of BST AVP mRNA-expressing cells, significantly greater than neonatally castrated males and control females, which were statistically similar (F(2,32)=5.85, P≤0.007); no MA differences were found.
  • Exogenous androgens did not maintain or masculinize AVP. Daily 1000 g TP during the first postnatal week did not maintain high AVP-ir in neonatally castrated males (Exp 3; neonatal castration reduced LS AVP-ir, F(2,26)=16.33, P≤0.0001; LHb sham vs castrated-oil P=0.053). Perinatal TP (75, 500, or 1000 g), T (100 g), or DHT (200 g) did not masculinize female LS or LHb AVP-ir; 1000 g TP reduced it in some cases. However, 1000 g TP lengthened anogenital distance, indicating biological activity.
  • Estrogenic manipulations affected AVP. Neonatal tamoxifen reduced AVP-ir in the male LS (t18=3.47, P≤0.003) and tended to reduce it in the LHb (P=0.061). Neonatal DES increased septal AVP-ir in females (U=12.50, P≤0.001) but did not significantly affect the LHb; the DES effect in the LS varied with litter sex ratio (F(2,10)=4.98, P≤0.04), being more effective in females from litters with more males.
  • A single 500 g TP injection on postnatal d 3 did not masculinize female AVP to male levels but increased it relative to oil-treated females in the LS (t14=2.36, P≤0.04) and LHb (t14=2.24, P≤0.05); repeated spaced TP had no effect.

Methods

  • Subjects were F4 and F5 generation prairie voles (M. ochrogaster) from laboratory colonies. Experiments 1–8 used perinatal manipulations including neonatal castration/sham, prenatal flutamide plus ATD or oil, postnatal TP/T/DHT, tamoxifen, and DES; adults were gonadectomized or sham-operated and implanted with a 2.5-cm T-filled SILASTIC capsule, then killed 3 wk later.
  • Brains were processed for AVP immunocytochemistry (fiber density in LS, LHb, and PVT) or in situ hybridization (AVP mRNA-expressing cells in BST and MA). Slides were randomized and coded; image analysis used NIH IMAGE 1.44.
  • Analyses used one-way ANOVA with Bonferroni post hoc tests; Exp 4 also used two-way ANOVA for females; Exp 8 used unpaired t tests or Mann-Whitney U tests. Significance was P≤0.05.

Limitations

  • Stated: circulating T levels after the 2.5-cm SILASTIC capsule implantation were not determined and were likely supraphysiological; it is not known when the AVP system first becomes sexually dimorphic in prairie voles; the authors state they can only conjecture about species differences.
  • Stated/evident: some data were lost or excluded (e.g., missing BST/MA data for some subjects; outliers removed by Dixon’s test), and sample sizes were modest across experiments.
  • Evident: findings are limited to one vole species and to AVP-ir/mRNA measures; no direct neonatal hormone measurements are reported in this study except preliminary data cited in the Discussion, and possible dietary phytoestrogen differences are discussed as a potential confound.

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

Integrity signals

Registry

Crossref · 10.1210/en.2004-1315

Cites

71 of 71 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 51 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    Prenatal testosterone propionate treatment has no effect on AVP immunoreactivity in the lateral habenula of female prairie voles.

    “Similar to the LS, females that received postnatal TP showed a significant decrease in AVP-ir in the LHb (F (1,1) ϭ 9.72, P Յ 0.005), and no effects of prenatal TP (F (1,1) ϭ 0.34, P Ն 0.56) or prenatal by postnatal interaction (F (1,1) ϭ 0.01, P Ն 0.9) were found.”
    Quote in the paperSubstantial claim
  • Findingown data

    All groups of female prairie voles, regardless of perinatal testosterone propionate treatment, have significantly less AVP immunoreactivity in the lateral septum than control males.

    “All groups of females, regardless of perinatal treatment, had significantly less AVP-ir in the LS (F (4,39) ϭ 14.46, P Յ 0.0001; Fig. 6) and LHb (F (4,39) ϭ 12.849, P Յ 0.0001; Fig. 6) than control males.”
    Quote in the paperSubstantial claim
  • Findingown data

    Administration of testosterone, dihydrotestosterone, or either 75 g or 1000 g testosterone propionate for the first week of life has no effect on AVP immunoreactivity in the lateral habenula of female prairie voles.

    “These treatments also had no effect on AVP-ir in the LHb (F (5,45) ϭ 4.49, P Յ 0.003; Fig. 9).”
    Quote in the paperSubstantial claim
  • Findingown data

    Administration of testosterone, dihydrotestosterone, or either 75 g or 1000 g testosterone propionate for the first week of life has no effects on AVP immunoreactivity in the lateral septum of female prairie voles.

    “Administration of T, DHT, or either dose of TP for the first week of life had no effects on AVP-ir in the LS of females.”
    Quote in the paperSubstantial claim
  • Findingown data

    Neonatal castration reduces the number of AVP mRNA-expressing cells in the bed nucleus of the stria terminalis of male prairie voles.

    “Neonatal castration reduced the number of AVP mRNA-expressing cells in the bed nucleus of the stria terminalis and AVP immunoreactivity (ir) in the LS and LHb.”
    • Also stated in: § Results. · plain
      “Sham-castrated males had the greatest number of AVP mRNA-expressing cells, which was significantly greater than both neonatally castrated males and control females, which were statistically similar to each other.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The AVP mRNA riboprobe derived from the rat AVP sequence detects prairie vole AVP mRNA.
  • May combine two claims. 29
  • Classified as probably not a substantial claim. 13

  • May combine two claims.

    Findingown data

    Within the medial amygdala, there are no significant differences between groups in the number of AVP mRNA-expressing cells or in the number of grains per cell.

    “Within the MA, there were no significant differences between any of the groups in the number of AVP mRNAexpressing cells (F (2,34) ϭ 0.18, P Ն 0.8; Fig. 3) or in the number of grains per cell (F (2,34) ϭ 1.41, P Ն 0.2; data not shown).”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: The AVP mRNA riboprobe derived from the rat AVP sequence detects prairie vole AVP mRNA.

  • May combine two claims.

    Findingown data

    In the lateral septum, all female treatment groups except testosterone-treated females are significantly lower than control males; testosterone-treated females are not significantly different from any group.

    “All groups of females were statistically similar to each other and had significantly lower AVP-ir than control males, with the exception of T-treated females, which were not significantly different than any group (F (5,48) ϭ 2.51, P Յ 0.05).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Treating neonatally castrated male prairie voles with testosterone propionate during the first postnatal week does not maintain high levels of AVP immunoreactivity and nonsignificantly decreases it.

    “Surprisingly, daily injections of 1000 g of testosterone propionate (TP) during the first postnatal week did not maintain high levels of AVP-ir in neonatally castrated males.”
    • Also stated in: § Experiment 3: effects of T replacement on AVP-ir in neonatally castrated males · plain
      “However, treating neonatally castrated males with TP during the first week of life after neonatal castration did not maintain high levels of AVP-ir but, instead, nonsignificantly decreased AVP-ir (Fig. 5).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    1000 g testosterone propionate lengthened anogenital distance, indicating that testosterone propionate was biologically active.

    “However, 1000 g TP lengthened anogenital distance, indicating that TP was biologically active.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Extending testosterone propionate treatment past the first neonatal week has no effect on AVP immunoreactivity in female prairie voles, and both female groups have significantly lower AVP immunoreactivity than oil-treated males in the lateral septum and lateral habenula.

    “Extending TP treatment past the first neonatal week had no effect on AVP-ir in females, with both groups of females having significantly lower AVP-ir than oil-treated males in the LS (F (2,21) ϭ 9.31, P Յ 0.002) and LHb (F (2,21) ϭ 6.54, P Յ 0.007; Fig. 8).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In experiment 6, injection of 500 g TP on postnatal days 2, 5, 8, and 11 also did not affect anogenital distance, and the distance was shorter for both groups of females than males.

    “In Experiment 6, injection of 500 g TP on postnatal d 2, 5, 8, and 11 also did not affect anogenital distance, and the distance was shorter for both groups of females than males (F (2,19) ϭ 56.64, P Յ 0.0001).”
    Quote in the paperNot a substantial claim

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

    Findingown data

    Adult body weight of female prairie voles was not affected by any perinatal treatment in any experiment.

    “Adult body weight of females was not affected by any perinatal treatment in any experiment (all P Ն 0.5; data not shown).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    In experiment 7, 1 mg TP lengthened female anogenital distance compared with other groups of females, whereas 75 g TP, 100 g T, and 200 g DHT did not; anogenital distance in all groups of females was shorter than males.

    “In experiment 7, 1 mg TP lengthened female anogenital distance compared with other groups of females, whereas 75 g TP, 100 g T, and 200 g DHT did not; anogenital distance in all groups of females was shorter than males (F (5,45) ϭ 41.92, P Յ 0.0001).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingcitation

    Neonatal castration significantly reduced adult anogenital distance in male prairie voles in experiments 1-3, and TP treatment increased it to control values in neonatally castrated males in experiment 3.

    “As found previously (26), neonatal castration significantly reduced adult anogenital distance in males in experiments 1-3, and TP treatment increased it to control values in neonatally castrated males in experiment 3 (all P Յ 0.01; data not shown).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [26]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Findingown data

    In prairie voles, unlike rats, perinatal treatment with exogenous TP could not maintain masculine AVP expression in neonatally castrated males and did not masculinize this system in females.

    “The surprising finding was that, unlike rats, perinatal treatment with exogenous TP could not maintain masculine AVP expression in neonatally castrated males and did not masculinize this system in females.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In DES-treated female prairie voles, there was a significant polynomial regression between AVP-ir in the LS and the sex ratio of their litter at birth, with DES being more effective in masculinizing septal AVP-ir in females originating from litters with more males.

    “In addition, there was a significant polynomial regression between the levels of AVP-ir in the LS of DES-treated females and the sex ratio of their litter at birth (F (2,10) ϭ 4.98, P Յ 0.04), with DES being more effective in masculinizing septal AVP-ir in females originating from litters that had more males.”
    Quote in the paperNot a substantial claim

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

    Findingown data

    Adult body weight of male prairie voles was not affected by any perinatal treatment in any experiment.

    “Adult body weight of males was not affected by any perinatal treatment in any experiment (all P Ն 0.1; data not shown).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Prenatal and postnatal treatment with TP significantly increased anogenital distance in the subset of females examined from experiment 4, although anogenital distance of both TP-treated and oil-treated females was shorter compared with control males.

    “Also similar to results found previously (26), prenatal and postnatal treatment with TP significantly increased anogenital distance in the subset of females examined from experiment 4, although anogenital distance of both TP-treated and oil-treated females was shorter compared with control males (F (2,19) ϭ 100.1, P Յ 0.0001; data not shown).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [26]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Findingown data

    Neonatal castration of male prairie voles completely demasculinized AVP mRNA expression in the BST and partially demasculinized AVP-ir fiber density in the LS and LHb, similar to rats.

    “The present results demonstrate that neonatal castration of male prairie voles produced effects similar to those in rats, that is, completely demasculinizing AVP mRNA expression in the BST and partially demasculinizing AVP-ir fiber density in the LS and LHb.”
    • Also stated in: § Discussion · plain
      “although neonatal castration completely demasculinized AVP mRNA expression in the BST, it did not fully demasculinize AVP-ir in the terminal areas”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Preliminary data indicate that T is nondetectable in prairie voles of either sex when killed between 18-24 h after birth, whereas estradiol levels are higher in neonatal males than in females.

    “preliminary data indicate that T is nondetectable in prairie voles of either sex when killed between 18 -24 h after birth, whereas estradiol levels are higher in neonatal males than in females (Lonstein, J. S., and J. A. French, preliminary data).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Repeated administration of T, DHT, lower doses of TP, or the single injection of 500 g TP still did not masculinize the AVP system in females but was not feminizing.

    “Nonetheless, repeated administration of T, DHT, lower doses of TP, or the single injection of 500 g TP still did not masculinize the AVP system in females but was not feminizing.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Neonatal castration demasculinizes parental responsiveness in virgin male prairie voles, but perinatal treatment of virgin females with testosterone propionate does not masculinize it.

    “Furthermore, neonatal castration demasculinizes parental responsiveness in virgin male prairie voles, but perinatal treatment of virgin females with TP does not masculinize it (25,26).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [25]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [26]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

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

    Premisenone given

    The influence of gonadal hormones on the development of any sex difference in the prairie vole brain has not been examined.

    “The influence of gonadal hormones on the development of any sex difference in the prairie vole brain has not been examined and may differ from rats.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    In rats, the sexually dimorphic extrahypothalamic AVP system originates in AVP-synthesizing cells of the bed nucleus of the stria terminalis and medial amygdala and terminates densely in the lateral septum and lateral habenula.

    “In rats, this sexually dimorphic system originates in AVP-synthesizing cells of the bed nucleus of the stria terminalis (BST) and medial amygdala (MA) and terminates densely in the lateral septum (LS) and lateral habenula (LHb) (7,8).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [7]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [8]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Premisecitation

    In rats, testosterone propionate requires aromatization into estradiol for much of its effect on extrahypothalamic AVP expression, although high doses of dihydrotestosterone can also partly masculinize this system.

    “TP apparently requires aromatization into estradiol for much of its effect (14), although the reduced androgen dihydrotestosterone (DHT) can also partly masculinize this system in rats if given in high doses (14).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [14]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Premisecitation

    Unlike rats, neonatal castration does not demasculinize sexual behavior in male prairie voles and, in some cases, neonatal testosterone propionate does not masculinize it in females.

    “Unlike rats (22), neonatal castration does not demasculinize sexual behavior in male prairie voles and, in some cases, neonatal TP does not masculinize it in females (23,24).”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [22]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [23]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [24]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

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

    Premisecitation

    In rats, neonatal castration prevents masculinization of AVP expression, whereas treating neonatal females with testosterone propionate masculinizes it.

    “For example, neonatal castration of male rats prevents the masculinization of AVP expression, whereas treating neonatal females with testosterone propionate (TP) masculinizes it (12)(13)(14).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites (12)(13)(14)

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Interpretationown data

    This is the first example of a demasculinization of the male rodent brain after neonatal castration without a restoration after exogenous androgen treatment, and the first demonstration that perinatal androgens are ineffective in masculinizing a sexually dimorphic neural trait in a female rodent.

    “As far as we are aware, this is the first example of a demasculinization of the male rodent brain after neonatal castration without a restoration after exogenous androgen treatment, as well the first demonstration that perinatal androgens are ineffective in masculinizing a sexually dimorphic neural trait in a female rodent.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Numerous other species, including domestic pigs, hamsters, ferrets, and some primates, respond to perinatal manipulations in gonadal hormones differently than laboratory rats, demonstrating that sexual differentiation occurs differently across species.

    “Numerous other species, including domestic pigs, hamsters, ferrets, and some primates, respond to perinatal manipulations in gonadal hormones differently than laboratory rats (51), demonstrating that sexual differentiation occurs differently across species.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [51]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Classified as probably not a substantial claim.

    Study design and conductown data

    The total number of AVP mRNA-expressing cells in the bed nucleus of the stria terminalis and medial amygdala was quantified by a single observer unaware of subject condition.

    “The total number of AVP mRNA-expressing cells in six hemisections through the BST and MA were quantified by eye under bright-field illumination by a single observer (J.S.L.) unaware of subject condition.”
    Quote in the paperSubstantial claim

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

    Study design and conductown data

    Surrogate dams readily accepted foster pups, and fostered pups had very high survival rates.

    “surrogate dams readily accepted foster pups, and the fostered pups in all cases had very high survival rates.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    All slides were randomized and coded for analysis.

    “All slides were randomized and coded for analysis.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    In rats, castration of neonatal males before postnatal day 7 completely demasculinizes AVP mRNA expression in the BST and AVP-ir fiber density in the LS, whereas a single or repeated injection of TP to neonatally castrated males within the first 3 weeks of life maintains masculinization of AVP mRNA expression in the BST and partially maintains masculinization of AVP-ir fibers in the LS.

    “Castration of neonatal males (before postnatal d 7) completely demasculinizes AVP mRNA expression in the BST, as well as AVP-ir fiber density in the LS (12,14,48), whereas a single or repeated injection of TP to neonatally castrated males within the first 3 wk of life maintains masculinization of AVP mRNA expression in the BST and partially maintains masculinization of AVP-ir fibers in the LS (12,”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [12]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [14]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [48]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

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

    Premisecitation

    Treatment of perinatal females with androgens or estrogens significantly masculinizes these neural characteristics.

    “Conversely, most of these studies have demonstrated that treatment of perinatal females with androgens or estrogens significantly masculinizes these neural characteristics.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    In female rats, injection of TP relatively soon after birth completely masculinizes AVP-ir fiber density in the LS, and either a single or repeated injection of TP or estradiol benzoate during this time masculinizes the number of AVP mRNA-expressing cells in the BST.

    “In females, injection of TP relatively soon after birth completely masculinizes AVP-ir fiber density in the LS (48), and either a single or repeated injection of TP or estradiol benzoate during this time masculinizes the number of AVP mRNA-expressing cells in the BST (12,14).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [48]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [12]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [14]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Interpretationown data

    The findings that neonatal castration demasculinized AVP expression in males and that neonatal estrogenic manipulations affected AVP in both sexes suggest that at least part of the critical period for AVP sexual differentiation exists during early postnatal life.

    “Furthermore, our findings that neonatal castration demasculinized AVP expression in males and that neonatal estrogenic manipulations affected AVP in both sexes suggest that at least part of this critical period exists during early postnatal life.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    The AVP system probably becomes sexually dimorphic early in development in prairie voles, and it is likely that the TP treatments encompassed this period.

    “It is not known when this system first becomes sexually dimorphic in prairie voles, but considering that prairie vole pups are precocial compared with rat pups and, at birth or within a few days after, have teeth and fur and are mobile (53-55), this system probably becomes sexually dimorphic early in development in voles, and it is likely that our TP treatments encompassed this period.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [53]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [54]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [55]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

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

    Study design and conductown data

    Survival of neonatally manipulated pups raised by their own dams was invariably 100% in all experiments.

    “survival of neonatally manipulated pups raised by their own dams was invariably 100% in all experiments.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    In virgin prairie voles, parental behavior shows the same pattern: males are demasculinized by neonatal castration but the behavior of females is not masculinized by perinatal TP.

    “Interestingly, the same effect can be found for sex differences in the parental behavior of virgin prairie voles, such that males are demasculinized by neonatal castration but the behavior of females is not masculinized by perinatal TP (26).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [26]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Interpretationown data

    Estrogen is a major masculinizing factor of the AVP system in prairie voles, similar to rats.

    “This indicates that, similar to rats, estrogen is a major masculinizing factor of this system in prairie voles.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Early exposure to phytoestrogens modulates reproductive development and endocrine function in rodents and may or may not influence sexual differentiation of their brain.

    “Early exposure to phytoestrogens modulates reproductive development and endocrine function in rodents (64,65) and may (66) or may not (64, 67) influence sexual differentiation of their brain.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [64]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [65]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [66]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

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

    Premisecitation

    Gonadal hormones can down-regulate estradiol receptors in some areas of the developing and adult rat brain.

    “Gonadal hormones can down-regulate estradiol receptors in some areas of the developing and adult rat brain (56,57)”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [56]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [57]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

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

    Interpretationreasoning

    Perinatal hormone manipulations may influence sexual differentiation in rats and voles differently because of inadvertent differences in endocrine factors present in the diets of their mothers.

    “Therefore, perinatal hormone manipulations may influence sexual differentiation in rats and voles differently because of inadvertent differences in endocrine factors present in the diets of their mothers.”
    Quote in the paperNot a substantial claim
38 of 38

  • The paper relies on this without support.

    The AVP mRNA riboprobe derived from the rat AVP sequence detects prairie vole AVP mRNA.

    “The riboprobe contained 293 bases that complemented the rat AVP mRNA.”

  • The authors state this as an assumption.

    Circulating testosterone levels in adult prairie voles after implantation of a 2.5-cm long capsule are likely supraphysiological.

    “Circulating levels of T in adult voles after implantation of a 2.5-cm long capsule have not been determined but are likely supraphysiological.”
2 of 2

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?

  • Neonatal manipulation of estradiol or testosterone alters prairie vole affiliative behaviors later in life, and estradiol administration during adulthood alters estrus and locomotor activity.

    “Similarly, neonatal manipulation of estradiol or testosterone alters prairie vole affiliative behaviors later in life, and estradiol administration during adulthood alters estrus and locomotor activity (57, 128, 129).”

    From Endocrine Disruption of Vasopressin Systems and Related Behaviors (2017) · cites this paper as [129]

    The introductory passage (page unavailable) reports that neonatal castration changes later parental responsiveness in male prairie voles, supporting a limited link between neonatal hormone manipulation and affiliative behavior. It also says perinatal testosterone propionate does not masculinize parental responsiveness in females. The paper examines hormone effects on vasopressin expression; it does not report that adult estradiol administration alters estrus or locomotor activity. (LLM, conf=0.84)

    Experimental — accuracy not yet measured

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.

  • Experiment 2 MA ANOVA implies more animals than enrolled

    Minor

    The listed groups total 36 animals, and MA data were unavailable for one sham male. That leaves 35 observations and 32 residual degrees of freedom for a three-group ANOVA; the reported 34 would require 37 observations.

  • DES-group regression degrees of freedom imply too many subjects

    Minor

    The regression is described as using DES-treated females, of whom there were 11. A polynomial regression with two model degrees of freedom on 11 observations would have 8 residual degrees of freedom, not the reported 10.

  • Experiment 8 female LHb result uses a test the methods say was replaced

    Minor

    The data-analysis methods say female experiment 8 data were analyzed with a Mann-Whitney U test instead of an unpaired t test, yet the female LHb result reports a t test.

  • Experiment 2 BST degrees of freedom do not reflect exclusions

    Minor

    The stated groups total 36 animals. Four BST observations were excluded or unavailable, leaving 32 animals and 29 residual degrees of freedom for a three-group ANOVA, rather than the reported 32.

  • Experiment 1 ANOVA degrees of freedom exceed the sample sizes

    Minor

    The three groups contain 30 animals in total. A three-group ANOVA would have 27 residual degrees of freedom for the LHb, not 28; removing the stated LS outlier leaves 29 animals and 26 residual degrees of freedom, not 27.

  • Experiment 4 two-way ANOVA reports one residual degree of freedom

    Minor

    The four female groups contain 35 animals. A two-way ANOVA with the stated prenatal and postnatal factors would have 31 residual degrees of freedom, not the reported 1; the same F(1,1) format is used for the other effects.

6 of 6

Cites

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Cited by

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Related

Authors
Lonstein, J. S., Rood, B. D., & De Vries, G. J.
Affiliations
  • Michigan State University (US)
  • University of Massachusetts Amherst (US)
Corresponding author
Lonstein, Joseph S.
ORCID
Venue
Endocrinology
Publisher
The Endocrine Society
Published
2005-03
Volume
146
Issue
3
Pages
1559-1567
ISSN
  • 0013-7227 (ISSN-L)
  • 1945-7170
DOI
10.1210/en.2004-1315
PMC
PMC1444947
OpenAlex
W2056913049
Cited by
33
References
71
Citations by year
2025: 1 · 2022: 1 · 2021: 1 · 2019: 1 · 2018: 1 · 2017: 1 · 2014: 4 · 2013: 1 · 2012: 4
FWCI
4.73
Type
journal article
Language
en
Open access
Locations
  • Endocrinology (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (submittedVersion)
  • ScholarWorks@UMassAmherst (University of Massachusetts Amherst) (submittedVersion)
Funders
  • National Institute of Mental Health (K02 MH001497, MH47538, R01-MH047538)
  • Eunice Kennedy Shriver National Institute of Child Health and Human Development (HD08392, F32 HD008392)
Topics
  • Social Sciences › Psychology › Social Psychology › Neuroendocrine regulation and behavior
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Sexual Differentiation and Disorders
  • Health Sciences › Medicine › Reproductive Medicine › Hypothalamic control of reproductive hormones
Keywords
vasopressin, sexual differentiation, Microtus ochrogaster, bed nucleus of the stria terminalis, lateral septum, lateral habenula, testosterone propionate, neonatal castration, tamoxifen, diethylstilbestrol, estrogen receptor antagonism, anogenital distance
MeSH terms
Animals, Arginine Vasopressin, Body Weight, Arvicolinae, Diethylstilbestrol, Dose-Response Relationship, Drug, Female, Hypothalamus, Immunohistochemistry, Male, RNA, Messenger, Sex Differentiation, Sex Factors, Tamoxifen, Testosterone, In Situ Hybridization, Gonadal Hormones, Testosterone Propionate, Rats

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.

Statistics and arithmetic are recomputed, not judged.

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