CoChat
Sign Up
Cited by 167Neuroendocrine regulation and behaviorNeuropeptides and Animal PhysiologyMemory and Neural Mechanisms

Oxytocin enhances observational fear in mice

Open access
Pisansky, Marc T., Hanson, Leah R., Gottesman, Irving I., Gewirtz, Jonathan C.·2017·Nature Communications
DOI 10.1038/s41467-017-02279-5PMC PMC5727393
Open this paper in CoChat — free accountOpen PDF

Abstract

Empathy is fundamental to human relations, but its neural substrates remain largely unknown. Here we characterize the involvement of oxytocin in the capacity of mice to display emotional state-matching, an empathy-like behavior. When exposed to a familiar conspecific demonstrator in distress, an observer mouse becomes fearful, as indicated by a tendency to freeze and subsequent efforts to escape. Both intranasal oxytocin administration and chemogenetic stimulation of oxytocin neurons render males sensitive to the distress of an unfamiliar mouse. Acute intranasal oxytocin penetrates the brain and enhances cellular activity within the anterior cingulate cortex, whereas chronic administration produces long-term facilitation of observational fear and downregulates oxytocin receptor expression in the amygdala. None of these manipulations affect fear acquired as a result of direct experience with the stressor. Hence, these results implicate oxytocin in observational fear in mice (rather than fear itself) and provide new avenues for examining the neural substrates of empathy.

AI snapshot

Written from the paper’s full text.

TL;DR

Oxytocin enhances observational/socially transmitted fear in mice, while leaving directly acquired Pavlovian fear largely unaffected. Acute and chronic intranasal oxytocin, as well as chemogenetic activation of hypothalamic oxytocin neurons, increase freezing in unfamiliar male observers, whereas an oxytocin receptor antagonist reduces it.

Key findings

  • Familiar male observers froze significantly more than unfamiliar male observers (two-tailed t-test: t(25)=2.96, p=0.008), while familiar and unfamiliar females did not differ (t(19)=0.728, p=0.48).
  • Acute intranasal oxytocin (20 μg/kg) elevated CSF oxytocin (t(19)=2.73, p=0.015) and enhanced freezing in unfamiliar males (t(23)=2.49, p=0.020); chronic oxytocin (20 μg/kg, 5 daily doses) also enhanced freezing 10 days after cessation (t(21)=2.64, p=0.016) and downregulated Oxtr in the CeL (t(19)=2.08, p=0.05) but not ACC (p=0.95).
  • Chemogenetic activation of PVN oxytocinergic neurons with rM3D(Gs) enhanced freezing in unfamiliar males (ANOVA: F2,28=3.41, p=0.048), and the oxytocin receptor antagonist L-368,899 dose-dependently reduced freezing in familiar males (ANOVA: F2,35=5.73, p=0.007).
  • Oxytocin manipulations did not affect direct Pavlovian fear acquisition, expression, or extinction, and I125-labeled oxytocin was detected in brain regions including ACC, amygdala, and hypothalamus after intranasal delivery.

Methods

  • C57/B6 mice, 8–12 weeks old, tested as same-sex observer–demonstrator pairs; social fear was measured as observer freezing during demonstrator foot-shock conditioning (0.8 mA, 1.5 s, 15 trials), with a separate socially induced avoidance paradigm for escape behavior.
  • Pharmacological and chemogenetic manipulations included intranasal oxytocin (20 μg/kg acute or 5 daily doses), systemic L-368,899 (5 or 10 mg/kg I.P.), and CNO (3 mg/kg I.P.) in Oxt-IRES-Cre mice with bilateral PVN injections of AAV2-hSyn-DIO-rM3D(Gs)-mCherry.
  • Analyses used two-tailed Student’s t-tests, one-way ANOVA, and Dunnett’s post hoc tests, with significance set at P<0.05; supporting measures included CSF ELISA, I125-oxytocin distribution, qRT-PCR for Oxtr, and c-Fos immunohistochemistry.

Limitations

  • Stated: I125-radiolabeling cannot distinguish intact oxytocin from labeled peptide fragments; ELISA could not extract/quantify oxytocin from brain tissue; the experiments did not disambiguate relatedness from familiarity; and no statistical methods were used to predetermine sample sizes.
  • Evident: the key oxytocin manipulations were conducted primarily in male mice, so sex-specific generalization of the oxytocin effects is not established by the reported experiments.

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

Integrity signals

Registry

Crossref · 10.1038/s41467-017-02279-5

Cites

84 of 84 references verified0 citation errors · 0 not checkable

Statistics

21 tests recomputed · 0 with the reported and recomputed p on opposite sides of .05

Claims the paper makes

0 of 24 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    Acute intranasal oxytocin elevates oxytocin levels in cerebrospinal fluid in behaviorally naive mice.

    “Intranasal oxytocin successfully elevated CSF levels in behaviorally naive mice, thereby confirming the entry of oxytocin into brain CSF (Fig. 3b).”
    • Also stated in: § Intranasal oxytocin enhances social fear · plain
      “In behaviorally naïve mice administered acute intranasal oxytocin (20 μg/kg; dark blue, n = 10), concentrations of the neuropeptide in CSF were significantly elevated compared to saline-treated controls (gray, n = 10) (two-tailed Student’s t-test: t(19) = 2.73, p = 0.015).”
    Quote in the paperSubstantial claim
  • Findingown data

    Familiar and unfamiliar female mice do not differ significantly in levels of freezing during demonstrator conditioning.

    “Familiar (black, n = 10) and unfamiliar (gray, n = 10) female mice did not exhibit significant differences in levels of freezing (two-tailed Student’s t-test: t(19) = 0.728, p = 0.48)”
    • Also stated in: § Individual differences in social fear · plain
      “In female mice, we discerned no effect of familiarity—that is, sibling (henceforth, familiar) and non-sibling/non-cagemate (henceforth, unfamiliar) observers exhibited high and comparable levels of freezing during demonstrator conditioning (Fig. 1b, right).”
    Quote in the paperSubstantial claim
  • Findingown data

    Unfamiliar male observer mice exhibit significantly less freezing than familiar male observers during demonstrator conditioning.

    “Unfamiliar male observers (gray, n = 13) exhibited significantly less freezing compared to familiar male observers (black, n = 13) across demonstrator conditioning trials (repeated measure ANOVA: F 1,390 = 6.00, p < 0.0001), and when averaged (two-tailed Student’s t-test: t(25) = 2.96, p = 0.008).”
    • Also stated in: § Individual differences in social fear · plain
      “In contrast, unfamiliar male observers exhibited a significant deficit in freezing compared to familiar observers—both when quantified across conditioning trials (Fig. 1b, left) and when averaged (Fig. 1b, right).”
    Quote in the paperSubstantial claim
  • Findingown data

    Chronic intranasal oxytocin (20 μg/kg, 5 daily doses) significantly enhances freezing in unfamiliar male observers compared to saline-treated controls.

    “or chronic (20 μg/kg, 5 daily doses; light blue, n = 11; two-tailed Student’s t-test: t(21) = 2.64, p = 0.016) intranasal oxytocin exhibited a significant enhancement of freezing compared to saline-treated controls (gray, n = 12/11, respectively).”
    • Also stated in: § Intranasal oxytocin enhances social fear · plain
      “Similar to acute oxytocin, this regimen produced a significant enhancement of freezing in oxytocin-treated unfamiliar observers, compared to saline-treated controls (Fig. 3c).”
    Quote in the paperSubstantial claim
  • Findingown data

    Acute intranasal oxytocin (20 μg/kg, single dose, 30 min prior) significantly enhances freezing in unfamiliar male observers compared to saline-treated controls.

    “Unfamiliar male observers given acute (20 μg/kg, single dose, 30 min prior; dark blue, n = 12; two-tailed Student’s t-test: t(23) = 2.49, p = 0.020) or chronic (20 μg/kg, 5 daily doses; light blue, n = 11; two-tailed Student’s t-test: t(21) = 2.64, p = 0.016) intranasal oxytocin exhibited a significant enhancement of freezing compared to saline-treated controls (gray, n = 12/11, respectively).”
    • Also stated in: § Intranasal oxytocin enhances social fear · plain
      “For unfamiliar male mice, this regimen produced a significant enhancement of freezing compared to saline-treated controls (Fig. 3c).”
    Quote in the paperSubstantial claim
  • May combine two claims. 17
  • The quote only partly states this claim. 1
  • Classified as probably not a substantial claim. 23

  • May combine two claims.

    Findingown data

    Acute intranasal oxytocin given prior to conditioning does not alter acquisition, expression, or extinction of directly acquired fear.

    “We did not note any differences in these measures when intranasal oxytocin was given acutely prior to conditioning (Supplementary Fig. 8). Therefore, the effects of oxytocin on socially acquired fear were not secondary to alterations in the capacity for fear conditioning in general.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Initial locomotor activity (M1) is equivalent across groups, but unfamiliar male mice fail to maintain freezing behavior throughout each conditioning trial (M2-12).

    “Initial locomotor activity (M1) was equivalent across groups, indicating similar orientation responses for all observers; however, unfamiliar male mice failed to maintain freezing behavior throughout each conditioning trial (M2-12) (Fig. 1e).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In the socially induced avoidance paradigm, both familiar and unfamiliar observer mice exhibit significantly more time on the non-demonstrator side during acclimation on the second conditioning day compared to non-conditioned controls, but only familiar observers maintain this behavior during conditioning trials.

    “During acclimation on this second conditioning day, both familiar and unfamiliar observer mice exhibited significantly more time on the non-demonstrator side compared to non-conditioned controls. However, only familiar observers maintained this behavior during conditioning trials (Fig. 2b).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Chronic intranasal oxytocin significantly reduces oxytocin receptor (Oxtr) transcript expression in the lateral division of the central nucleus of the amygdala (CeL) but not in the ACC.

    “Chronic intranasal oxytocin significantly reduced oxytocin receptor (Oxtr) transcript expression in the lateral division of the central nucleus of the amygdala (CeL) (two-tailed Student’s t-test: t(19) = 2.08, p = 0.05), but not the ACC (two-tailed Student’s t-test, t(19) = 0.06, p = 0.95), assayed one day after completion of chronic oxytocin treatment.”
    • Also stated in: § Intranasal oxytocin enhances social fear · plain
      “amygdala tissue recovered from oxytocin-treated mice showed a significant downregulation of the Oxtr transcript, whereas there was no difference in the ACC (Fig. 3e, Supplementary Fig. 11).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Systemic administration of the oxytocin receptor antagonist L-368,899 hydrochloride to familiar male mice produces a dose-dependent reduction in freezing compared to saline-treated controls, but no differences in direct acquisition of Pavlovian fear.

    “Familiar male observers systemically administered the oxytocin receptor antagonist L-368,899 hydrochloride (OXTA; I.P. single dose, 30 min prior; red, n = 12 per group) showed dose-dependent reductions in socially transmitted fear at 5 mg/kg and 10 mg/kg, compared to saline-treated controls (ANOVA: F 2,35 = 5.73, p = 0.007).”
    • Also stated in: § Systemic oxytocin receptor antagonism reduces social fear · plain
      “Administration of the OXTA 30 min prior to demonstrator conditioning (Fig. 3a) produced a dose-dependent reduction in freezing compared to saline-treated controls (Fig. 3d), but no differences in direct acquisition of Pavlovian fear in these same observer mice (Supplementary Fig. 12a).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: L-368,899 hydrochloride specifically antagonizes oxytocin receptors in vivo at the doses used.

  • May combine two claims. The quote only partly states this claim.

    Findingown data

    Only familiar male observers exhibit a significant preference for the non-demonstrator side at the post-test and a conditioned avoidance of the demonstrator-containing side.

    “only familiar male observers exhibited a significant preference for the non-demonstrator side at the post-test, measured on the day following the last conditioning day (Fig. 2c). Finally, a normalized score of avoidance indicated that only familiar observer mice exhibited a conditioned avoidance of the demonstrator-containing side (Fig. 2d).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Acute intranasal oxytocin significantly enhances the percentage of c-Fos+/DAPI cells within the anterior cingulate cortex but not the primary motor area M1.

    “Using an immunohistochemical approach, we measured elevated c-Fos protein expression in the ACC, but not the neighboring primary motor region (M1), an area devoid of oxytocinergic inputs36 (Fig. 3d).”
    • Also stated in: § Intranasal oxytocin enhances social fear · plain
      “Acute intranasal oxytocin significantly enhanced the percentage of c-Fos+/DAPI cells within the ACC (two-tailed Student’s t-test: t(9) = 2.36, p = 0.048) but not M1 (two-tailed Student’s t-test: t(9) = 0.96, p = 0.369) (n = 5 mice per group).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [36] Knobloch, H. S. et al. Evoked axonal oxytocin release in the central amygdala attenuates fear response. Neuron 73, 553–566 (2012). source

      Not checked in the free run.

  • May combine two claims.

    Findingown data

    Familiar male observers raised in smaller litters exhibit higher freezing levels than those raised in larger litters, whereas unfamiliar male mice show the reverse relationship.

    “familiar observers raised in smaller litters (2–6 pups) exhibited higher freezing levels compared to those raised in larger litters (7–12 pups), whereas unfamiliar mice showed the reverse relationship (Fig. 1d).”
    • Also stated in: § Individual differences in social fear · plain
      “Socially transmitted fear differed between familiar (black; small: n = 3, large: n = 10) and unfamiliar (gray; small: n = 2, large: n = 11) male mice when the observer was reared within a relatively small litter (small: 2–6 pups; large: 7–12 pups)”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    I125-labeled oxytocin administered intranasally is detected robustly within the blood and olfactory bulbs and is distributed throughout the brain, including the anterior olfactory nucleus, anterior cingulate cortex, nucleus accumbens, amygdala, and hypothalamus.

    “I125-labeled oxytocin was detected robustly within the blood and olfactory bulbs, and was distributed throughout the brain, including anterior olfactory nucleus, anterior cingulate cortex (ACC), nucleus accumbens, amygdala, and hypothalamus (Table 1).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Saline-treated familiar male mice exhibit a significant correlation between socially and directly acquired freezing levels, a relationship that is not recapitulated in OXTA-treated mice.

    “Indeed, saline-treated familiar male mice exhibited a significant correlation between socially and directly acquired freezing levels, a relationship that was not recapitulated in OXTA-treated mice (Supplementary Fig. 12b).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: L-368,899 hydrochloride specifically antagonizes oxytocin receptors in vivo at the doses used.

  • May combine two claims.

    Findingown data

    DREADD activation does not cause differences in demonstrator vocalizations or in acquisition or retrieval of directly acquired fear.

    “There were no significant differences in demonstrator vocalizations (Supplementary Fig. 13), nor did DREADD activation cause differences in acquisition or retrieval of directly acquired fear (Supplementary Fig. 14).”
    Quote in the paperSubstantial claim

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

    Findingnone given

    Neither the ELISA kit nor a second kit proved effective for extracting and quantifying oxytocin contained within brain tissue.

    “Neither the aforementioned ELISA kit (Enzo Life Sciences, Inc.) nor a second kit (#K048, Arbor Assays, Inc.) proved effective for extracting and quantifying oxytocin contained within brain tissue.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    Both intranasal oxytocin administration and chemogenetic stimulation of oxytocin neurons render males sensitive to the distress of an unfamiliar mouse.

    “Both intranasal oxytocin administration and chemogenetic stimulation of oxytocin neurons render males sensitive to the distress of an unfamiliar mouse.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Socially and directly acquired fear are subject to individual differences along a common trait of fearfulness, and oxytocin neurotransmission contributes exclusively to the former.

    “Collectively, these findings support the idea that socially and directly acquired fear are subject to individual differences along a common trait of fearfulness41 and that oxytocin neurotransmission contributes exclusively to the former.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [41] Vaidyanathan, U. M. A., Patrick, C. J. & Bernat, E. M. Startle reflex potentiation during aversive picture viewing as an indicator of trait fear. Psychophysiology 46, 75–85 (2009). source

      Not checked in the free run.

    • Rests on an assumption: L-368,899 hydrochloride specifically antagonizes oxytocin receptors in vivo at the doses used.

  • May combine two claims.

    Interpretationown data

    The oxytocin system is both necessary and sufficient for the social transmission of fear.

    “Together, these findings indicate that the oxytocin system is both necessary and sufficient for the social transmission of fear.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: L-368,899 hydrochloride specifically antagonizes oxytocin receptors in vivo at the doses used.

  • Classified as probably not a substantial claim. May combine two claims.

    Premisecitation

    Prior studies have suggested that oxytocin is anxiolytic or attenuates measures of fear conditioning.

    “The results of a number of other studies have suggested that oxytocin is anxiolytic19–21 or attenuates measures of fear conditioning15–18.”
    Quote in the paperSubstantial claim0 of 7 sources checked
    • cites [19] Ring, R. H. et al. Anxiolytic-like activity of oxytocin in male mice: behavioral and autonomic evidence, therapeutic implications. Psychopharmacology 185, 218–225 (2006). source

      Not checked in the free run.

    • cites [20] Waldherr, M. & Neumann, I. D. Centrally released oxytocin mediates mating-induced anxiolysis in male rats. Proc. Natl Acad. Sci. USA 104, 16681–16684 (2007). source

      Not checked in the free run.

    • cites [21] Yoshida, M. et al. Evidence that oxytocin exerts anxiolytic effects via oxytocin receptor expressed in serotonergic neurons in mice. J. Neurosci. 29, 2259–2271 (2009). source

      Not checked in the free run.

    • cites [15] Viviani, D. et al. Oxytocin selectively gates fear responses through distinct outputs from the central amygdala. Science 333, 104–107 (2011). source

      Not checked in the free run.

    • cites [16] Lahoud, N. & Maroun, M. Oxytocinergic manipulations in corticolimbic circuit differentially affect fear acquisition and extinction. Psychoneuroendocrinology 38, 2184–2195 (2013). source

      Not checked in the free run.

    • cites [17] Modi, M. E. et al Peripheral administration of long-acting peptide OT receptor agonist inhibits fear induced freezing. J. Pharmacol. Exp. Ther. 358, 164–172 (2016). source

      Not checked in the free run.

    • cites [18] Pagani, J. H., Lee, H. J. & Young, W. S. Postweaning, forebrain-specific perturbation of the oxytocin system impairs fear conditioning. Genes Brain Behav. 10, 710–719 (2011). source

      Not checked in the free run.

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

    Study design and conductnone given

    c-Fos+ neurons were quantified by a treatment-naive experimenter.

    “quantification of c-Fos+ neurons was performed across eight slices containing the PVN (~−0.85 mm to bregma) by a treatment-naive experimenter.”
    • Also stated in: § Histology and c-Fos immunohistochemistry · plain
      “For intranasal oxytocin experiments, quantification of c-Fos+ neurons was performed across five slices containing the Cg1/2 and M1 regions (~+1 mm to bregma) by a treatment-naive experimenter.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductown data

    Video footage was scored by trained, group-naive experimenters.

    “Video footage was collected during all sessions using infrared web cameras and scored over 2-min bins by trained, group-naive experimenters using Button Box 5.0 (Behavioral Research Solutions, LLC).”
    Quote in the paperSubstantial claim

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

    Interpretationcitation

    The presence of a home-cage conspecific at the time oxytocin is administered could have the incidental effect of ameliorating subsequent acquisition of fear.

    “Thus, the presence of a home-cage conspecific at the time that oxytocin is administered could have the incidental effect of ameliorating subsequent acquisition of fear.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    No statistical methods were used to predetermine sample sizes.

    “No statistical methods were used to predetermine sample sizes, although sample sizes were consistent with those from previous studies of this kind7,9,29.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [7] Chen, Q., Panksepp, J. B. & Lahvis, G. P. Empathy is moderated by genetic background in mice. PLoS ONE 4, e4387 (2009). source

      Not checked in the free run.

    • cites [9] Sanders, J., Mayford, M. & Jeste, D. Empathic fear responses in mice are triggered by recognition of a shared experience. PLoS ONE 8, e74609 (2013). source

      Not checked in the free run.

    • cites [29] Burkett, J. P. et al. Oxytocin-dependent consolation behavior in rodents. Science 351, 375–378 (2016). source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Study design and conductown data

    Observer freezing behavior was scored by a treatment-naive experimenter.

    “Videos were scored for observer freezing behavior by a treatment-naive experimenter using Button Box 5.0 (Behavioral Research Solutions, LLC) (Supplementary Figs. 2, 4).”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    Paired home-cage mice were randomly designated as observer or demonstrator prior to the start of testing.

    “Paired home-cage mice were randomly designated as observer or demonstrator prior to the start of testing.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Systemically administered l-368,899 hydrochloride reaches the brain and preferentially binds with oxytocin receptors.

    “Systemically administered l-368,899 hydrochloride is known to reach the brain, where it preferentially binds with oxytocin receptors75.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [75] Boccia, M. L., Goursaud, A. P. S., Bachevalier, J., Anderson, K. D. & Pedersen, Ca Peripherally administered non-peptide oxytocin antagonist, L368,899®, accumulates in limbic brain areas: a new pharmacological tool for the study of social motivation in non-human primates. Horm. Behav. 52, 344–351 (2007). source

      Not checked in the free run.

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

    Study design and conductnone given

    Four mice were removed from analysis due to escaping the testing apparatus in the socially induced avoidance paradigm.

    “Four mice were removed from analysis due to escaping the testing apparatus in the socially induced avoidance paradigm.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Demonstrator freezing during conditioning was measured in a randomly selected subset of videos of male familiar and unfamiliar pairs.

    “Demonstrator freezing during conditioning was measured in a randomly selected subset of videos of male familiar and unfamiliar pairs (Supplementary Fig. 4).”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Slices used for c-Fos immunohistochemistry in Figs. 3d and 4d were randomly selected.

    “Slices used for c-Fos immunohistochemistry in Figs. 3d, 4d were randomly selected.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Samples contaminated with blood were discarded.

    “Samples contaminated with blood were discarded.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Samples were run in duplicate on a spectrophotometer at 405 nm.

    “Samples were then reconstituted in assay buffer and run in duplicate on a spectrophotometer (Multiskan EX, Thermo Scientific, Inc.) at 405 nm.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    One mouse was removed from analysis due to viral injection misplacement in chemogenetic experiments.

    “One mouse was removed from analysis due to viral injection misplacement in chemogenetic experiments.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    Vocalization recordings were not collected for one familiar male, one unfamiliar female, and two unfamiliar males with acute intranasal saline due to technical error.

    “Vocalization recordings were not collected for one familiar male, one unfamiliar female, and two unfamiliar males with acute intranasal saline due to technical error.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Two mice were removed from analysis due to poor perfusion following I125-oxytocin administration.

    “Two mice were removed from analysis due to poor perfusion following I125-oxytocin administration.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    All animals used for chemogenetics experiments had their brains removed and processed to confirm viral injection location.

    “All animals used for chemogenetics experiments had their brains removed and processed to confirm viral injection location.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Mice without Pavlovian fear conditioning experience were used for quantification of c-Fos+ neurons.

    “Mice without Pavlovian fear conditioning experience were used for quantification of c-Fos+ neurons.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    An experienced experimenter confirmed the accuracy of automated call detection using the generated spectrogram.

    “An experienced experimenter confirmed the accuracy of automated call detection using the generated spectrogram.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductcitation

    Observer mice did not have experience with foot-shock prior to demonstrator conditioning.

    “Observer mice did not have experience with foot-shock (i.e., “priming”) prior to demonstrator conditioning (cf. 7–9).”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [7] Chen, Q., Panksepp, J. B. & Lahvis, G. P. Empathy is moderated by genetic background in mice. PLoS ONE 4, e4387 (2009). source

      Not checked in the free run.

    • cites [8] Kim, E. S. E. J., Covey, E. & Kim, J. J. Social transmission of fear in rats: the role of 22-kHz ultrasonic distress vocalization. PLoS ONE 5, e15077 (2010). source

      Not checked in the free run.

    • cites [9] Sanders, J., Mayford, M. & Jeste, D. Empathic fear responses in mice are triggered by recognition of a shared experience. PLoS ONE 8, e74609 (2013). source

      Not checked in the free run.

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

    Study design and conductnone given

    Three replicate PCR reactions were conducted for each sample.

    “We conducted three replicate PCR reactions for each sample.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    The foot-shock parameters were piloted to elicit reliable auditory and visual signs of distress from demonstrators.

    “These parameters were piloted to elicit reliable auditory and visual signs of distress (i.e., vocalizations and thigmotaxis/freezing behavior, respectively) from demonstrators.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    Videos of demonstrator freezing were randomly selected from data in Fig. 1b for Supplementary Fig. 4.

    “Videos of demonstrator freezing were randomly selected from data in Fig. 1b for Supplementary Fig. 4.”
    Quote in the paperNot a substantial claim
38 of 38

  • The paper relies on this without support.

    L-368,899 hydrochloride specifically antagonizes oxytocin receptors in vivo at the doses used.

    “we systemically administered the non-peptidergic oxytocin receptor antagonist (OXTA) l-368,899 hydrochloride to familiar male mice.”

  • The paper relies on this without support.

    The intranasal oxytocin doses and timing used in this study are sufficient to activate central oxytocin receptors that mediate social fear.

    “Acute intranasal oxytocin penetrates the brain and enhances cellular activity within the anterior cingulate cortex, whereas chronic administration produces long-term facilitation of observational fear and downregulates oxytocin receptor expression in the amygdala.”

  • The authors state this as an assumption.

    Vocalizations recorded during conditioning were elicited from demonstrators.

    “Vocalizations were presumed to be elicited from demonstrators based on coincidence with foot-shock onset/offset and previously identified characteristics of distress vocalizations elicited by foot-shock7.”

  • The paper relies on this without support.

    The data were normally distributed.

    “Data were normally distributed; therefore, parametric tests were used to compare groups.”

  • The paper relies on this without support.

    The oxytocin receptor antagonist L-368,899 hydrochloride is selective for oxytocin receptors.

    “For the systemic oxytocin receptor antagonist (OXTA), mice were briefly anesthetized with isoflurane and quickly administered the selective OXTA L-368,899 hydrochloride (5 or 10 mg/kg, I.P.).”
5 of 5

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?

Statistics that don’t add up

Each reported p-value recomputed from its test statistic and degrees of freedom.

  • Inconsistent p-value: t(25) = 2.96, p = .008

    Minor

    The paper reports p = .008 here; recomputing from the test statistic and its degrees of freedom gives p = .007 (two-tailed). The check cannot tell whether the statistic or the p-value is the misprint.

    Reported p = .008 · recomputed p = .007

  • Inconsistent p-value: t(21) = 2.64, p = .016

    Minor

    The paper reports p = .016 here; recomputing from the test statistic and its degrees of freedom gives p = .015 (two-tailed). The check cannot tell whether the statistic or the p-value is the misprint.

    Reported p = .016 · recomputed p = .015

  • Inconsistent p-value: t(24) = 2.25, p = .035

    Minor

    The paper reports p = .035 here; recomputing from the test statistic and its degrees of freedom gives p = .034 (two-tailed). The check cannot tell whether the statistic or the p-value is the misprint.

    Reported p = .035 · recomputed p = .034

  • Inconsistent p-value: t(7) = 2.65, p = .050

    Minor

    The paper reports p = .05 here; recomputing from the test statistic and its degrees of freedom gives p = .033 (two-tailed). The check cannot tell whether the statistic or the p-value is the misprint.

    Reported p = .05 · recomputed p = .033

  • Inconsistent p-value: t(9) = 2.36, p = .048

    Minor

    The paper reports p = .048 here; recomputing from the test statistic and its degrees of freedom gives p = .043 (two-tailed). The check cannot tell whether the statistic or the p-value is the misprint.

    Reported p = .048 · recomputed p = .043

  • Inconsistent p-value: t(4) = 4.38, p = .035

    Minor

    The paper reports p = .035 here; recomputing from the test statistic and its degrees of freedom gives p = .012 (two-tailed). The check cannot tell whether the statistic or the p-value is the misprint.

    Reported p = .035 · recomputed p = .012

  • Inconsistent p-value: t(9) = 0.96, p = .369

    Minor

    The paper reports p = .369 here; recomputing from the test statistic and its degrees of freedom gives p = .362 (two-tailed). The check cannot tell whether the statistic or the p-value is the misprint.

    Reported p = .369 · recomputed p = .362

  • Inconsistent p-value: t(19) = 2.73, p = .015

    Minor

    The paper reports p = .015 here; recomputing from the test statistic and its degrees of freedom gives p = .013 (two-tailed). The check cannot tell whether the statistic or the p-value is the misprint.

    Reported p = .015 · recomputed p = .013

8 of 8

Statistics recomputed

Every reported test result, recomputed from its test statistic and degrees of freedom. A result shows as not adding up only once an independent check has confirmed it.

  • Doesn’t add upt(25) = 2.96reported p = .008 · recomputed p = .007

    “t(25) = 2.96, p = .008”

  • Doesn’t add upt(24) = 2.25reported p = .035 · recomputed p = .034

    “t(24) = 2.25, p = .035”

  • Doesn’t add upt(4) = 4.38reported p = .035 · recomputed p = .012

    “t(4) = 4.38, p = .035”

  • Doesn’t add upt(19) = 2.73reported p = .015 · recomputed p = .013

    “t(19) = 2.73, p = .015”

  • Doesn’t add upt(21) = 2.64reported p = .016 · recomputed p = .015

    “t(21) = 2.64, p = .016”

  • Doesn’t add upt(9) = 2.36reported p = .048 · recomputed p = .043

    “t(9) = 2.36, p = .048”

  • Doesn’t add upt(9) = 0.96reported p = .369 · recomputed p = .362

    “t(9) = 0.96, p = .369”

  • Doesn’t add upt(7) = 2.65reported p = .05 · recomputed p = .033

    “t(7) = 2.65, p = .050”

  • Not confirmedt(25) = 3.7reported p = .002 · recomputed p = .001

    “t(25) = 3.70, p = .002”

  • Consistentt(19) = 0.728reported p = .48 · recomputed p = .475

    “t(19) = 0.728, p = .48”

  • Consistentt(25) = 3.96reported p = .001 · recomputed p < .001

    “t(25) = 3.96, p = .001”

  • Consistentt(21) = 2.9reported p = .011 · recomputed p = .009

    “t(21) = 2.90, p = .011”

    The paper states a correction for multiple tests

  • Consistentt(19) = 2.32reported p = .042 · recomputed p = .032

    “t(19) = 2.32, p = .042”

    The paper states a correction for multiple tests

  • Consistentt(21) = 4.09reported p = .001 · recomputed p < .001

    “t(21) = 4.09, p = .001”

  • Consistentt(19) = 0.14reported p = .889 · recomputed p = .890

    “t(19) = 0.14, p = .889”

  • Consistentt(21) = 2.73reported p = .017 · recomputed p = .013

    “t(21) = 2.73, p = .017”

    The paper states a correction for multiple tests

  • Consistentt(19) = 1.27reported p = .229 · recomputed p = .219

    “t(19) = 1.27, p = .229”

    The paper states a correction for multiple tests

  • Consistentt(21) = 2.17reported p = .052 · recomputed p = .042

    “t(21) = 2.17, p = .052”

    The paper states a correction for multiple tests

  • Consistentt(23) = 2.49reported p = .02 · recomputed p = .020

    “t(23) = 2.49, p = .020”

  • Consistentt(19) = 2.08reported p = .05 · recomputed p = .051

    “t(19) = 2.08, p = .05”

  • Consistentt(19) = 0.06reported p = .95 · recomputed p = .953

    “t(19) = 0.06, p = .95”

21 of 21

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.

  • CeL-specific receptor result conflicts with whole-amygdala sampling

    Major

    The result is attributed specifically to the lateral division of the central amygdala, but Methods says the RNA was obtained from the whole amygdala. The described assay therefore does not establish the stated CeL-specific change.

  • Antagonist result points to the wrong figure panel

    Minor

    The text directs readers to Fig. 3d for the antagonist-induced reduction in freezing, whereas the Fig. 3 legend identifies panel d as c-Fos labeling and panel f as the antagonist experiment.

  • Open and closed conditioning days are reversed in Methods

    Minor

    The Results say observers were confined on the first, closed day and could escape on the second, open day. Methods assigns the opposite labels to those two procedures.

3 of 3

Cites

Flagged first, then by citations

Cited by

Sorted by citations

Related

Authors
Pisansky, M. T., Hanson, L. R., Gottesman, I. I., & Gewirtz, J. C.
Affiliations
  • University of Minnesota (US)
  • HealthPartners (US)
Corresponding author
Gewirtz, Jonathan C.
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2017-12-13
Volume
8
Issue
1
Pages
2102-2102
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-017-02279-5
PMC
PMC5727393
OpenAlex
W2773566393
Semantic Scholar
b652d631208729cab94baef8651b26994e2fd359
Cited by
167
References
83
Citations by year
2026: 15 · 2025: 24 · 2024: 19 · 2023: 16 · 2022: 25 · 2021: 22 · 2020: 13 · 2019: 15 · 2018: 17 · 2017: 1
FWCI
18.02
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
University of Minnesota
Topics
  • Social Sciences › Psychology › Social Psychology › Neuroendocrine regulation and behavior
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuropeptides and Animal Physiology
  • Life Sciences › Neuroscience › Cognitive Neuroscience › Memory and Neural Mechanisms
Keywords
oxytocin, empathy, anterior cingulate cortex, amygdala, oxytocin receptor, chemogenetic stimulation, intranasal oxytocin, freezing response, escape behavior
MeSH terms
Administration, Intranasal, Amygdala, Animals, Brain, Escape Reaction, Fear, Female, Gyrus Cinguli, Humans, Male, Mice, Inbred C57BL, Oxytocin, Reflex, Startle, Receptors, Oxytocin

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 2026-10-09.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-09.

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.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review