Insular cortex mediates approach and avoidance responses to social affective stimuli

May combine two claims.

Stress altered ultrasonic vocalizations during social interactions: rising and trill calls were frequent during naive interactions but reduced during stressed interactions.

“Rising and Trill calls, which are emitted during and in anticipation of rewarding stimuli, were quite frequent during naive interactions but reduced during stressed interactions (Fig. 1J).”

May combine two claims.

Approximately 82% of rats tested with PN 30 conspecifics exhibited a preference for the stressed target, while only 21% of rats tested with PN 50 conspecifics exhibited preference for the stressed target.

“Approximately 82% (42 of 51) of rats tested with PN 30 conspecifics exhibited a preference for the stressed target, while only 21% (10 of 46) of rats tested with PN 50 conspecifics exhibited preference for the stressed target.”

May combine two claims.

In tests with PN 30 conspecifics, experimental rats spent more time interacting with the stressed than naive conspecifics after vehicle injection and less time interacting with the stressed than naive conspecific after Gö injection.

“In tests with PN 30 conspecifics, experimental rats spent more time interacting with the stressed than naive conspecifics after vehicle injection (p < 0.0001) and less time interacting with the stressed than naive conspecific after Gö injection (p = 0.0090).”

May combine two claims.

Intra-insular OT administration increased time spent interacting with naive PN 30 conspecifics and decreased time with naive PN 50 conspecifics.

“OT increased time spent interacting with the naive PN 30 conspecific (p < 0.0005) and decreased time with the naive PN 50 conspecific (p = 0.002, Fig. 5F).”

May combine two claims.

After OT application, action potentials were significantly smaller with decreased amplitude, rise rate, decay rate and longer duration (half-width).

“after OT application, action potentials were significantly smaller with decreased amplitude, rise rate, decay rate and longer duration (half-width)”

May combine two claims.

OT reduced mEPSC amplitude but had no effect on mEPSC frequency in deep layer insular cortex pyramidal neurons.

“OT reduced mEPSC amplitude (p = 0.004) but had no effect on mEPSC frequency.”

May combine two claims.

Optical treatment did not influence exploratory activity or general behavior in the SAP test.

“and did not influence exploratory activity or general behavior in the SAP test (Sup. Fig. 5).”

May combine two claims.

In the vehicle condition of the OTRa SAP experiment, 7 of 10 experimental rats exhibited a preference for stressed over naive juveniles and 7 of the 8 experimental rats exhibited a preference for naive over stressed adults.

“In the vehicle condition, 7 of 10 experimental rats exhibited a preference for stressed over naive juveniles and 7 of the 8 experimental rats exhibited a preference for naive over stressed adults (Fig. 5B–C).”

May combine two claims.

In tests with PN 50 conspecifics, experimental rats spent less time interacting with the stressed conspecific in the vehicle condition but equal time interacting with the stressed conspecific after Gö injection.

“In tests with PN 50 conspecifics, the experimental rats spent less time interacting with the stressed conspecific in the vehicle condition (p = 0.0057) but equal time interacting with the stressed conspecific after Gö injection (p = 0.1614).”

May combine two claims.

There was a significant Age by Drug by Affect interaction in the OTRa SAP experiment, with greater time spent investigating the stressed PN 30 conspecific and less time investigating the stressed PN 50 conspecific in the vehicle conditions, but more time interacting with the naive PN 30 conspecific and less time with the naive PN 50 conspecific in the OTRa condition.

“There was a significant Age by Drug by Affect interaction (p < 0.0001), with greater time spent investigating the stressed PN 30 conspecific (vs. the naive PN30) and less time investigating the stressed PN 50 conspecific (vs. the naïve PN50) in the vehicle conditions (ps = 0.001), but more time interacting with the naive PN 30 conspecific (p = 0.06) and less time with the naive PN 50 conspecific (”

May combine two claims.

OT induced a leftward shift in input/output curves with significantly larger fEPSP amplitude compared to baseline at stimuli from 2 to 5V in acute insular cortex slices.

“OT induced a leftward shift, with significantly larger fEPSP amplitude compared to baseline at stimuli from 2 to 5V with washout levels differing from OT beginning at 3V (ps < 0.0001).”

May combine two claims.

Data-driven community detection analysis of Fos immunoreactivity in 29 ROIs revealed a network with 2 modules, in which ROIs were more highly correlated within each module than between modules.

“A data-driven community detection analysis revealed a network with 2 modules (Fig. 6A), in which ROIs were more highly correlated within each module than between modules (maximized modularity quotient = .59).”

May combine two claims.

In one-on-one tests, OTRa microinjection reduced the time spent interacting with stressed PN 30 conspecifics and increased the time interacting with stressed PN 50 conspecifics.

“OTRa microinjection reduced the time spent interacting with the stressed PN 30 conspecifics (p = 0.0008) and increased the time interacting with stressed PN 50 conspecifics (p = 0.009) in the one-on-one tests (Fig. 5E).”

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

OT did not influence the AMPA:NMDA current ratio.

“However, OT did not influence AMPA:NMDA current ratio (data not shown).”

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

In a pilot experiment, OT had no effect on spontaneous inhibitory post-synaptic potentials.

“In a pilot experiment, no effect of OT was found on spontaneous inhibitory post-synaptic potentials (data not shown).”

Classified as probably not a substantial claim.

A 5 min test length was selected because pilot studies showed a reliable decrease in social behavior after the first 5 min of test.

“The 5 min test length was selected after pilot studies in which we observed a reliable decrease in social behavior after the first 5 min of test.”

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

In the recording conditions, very few large events indicative of spontaneous glutamate release occurred.

“However, in our recording conditions very few large events, indicative of spontaneous glutamate release, occurred”

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

Shock parameters (2 footshocks, 1 mA, 5 sec duration, 60 sec inter-shock interval) were sufficient to produce conditioned fear in the authors' laboratory.

“Shock occurred in a separate room and shock parameters were selected because they were sufficient to produce a conditioned fear in our laboratory (data not shown).”

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

A portion of rats did not exhibit the expected behavior towards stress conspecifics.

“we observed a portion of rats that did not exhibit the expected behavior towards stress conspecifics.”

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

OT can increase spike output in vivo.

“Indeed, OT can increase spike output in vivo44”

• cites [44] Moaddab, M., Hyland, B. I. & Brown, C. H. Oxytocin excites nucleus accumbens shell neurons in vivo. Mol. Cell. Neurosci. 68, 323–330 (2015). sourceNot checked yet

  Not checked in the free run.

The quote only partly states this claim.

Optogenetic silencing of insular cortex pyramidal neurons prevented the expression of social affective behaviors.

“Thus, silencing the insular cortex prevented the expression of social affective behaviors.”

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

Foot-shocked rats emit social odors that can either attract conspecifics in social buffering or serve as social alarm signals.

“Foot-shocked rats emit social odors that can either attract conspecifics in social buffering or serve as social alarm signals38.”

• cites [38] Kiyokawa, Y. Social odors: alarm pheromones and social buffering. Curr. Top. Behav. Neurosci. 30, 47–65 (2017). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Data-driven network analyses revealed functional integration among regions in the social brain, and suggest that the insula is positioned to interact with previously identified nodes important for social and affective behaviors.

“Thus, data-driven network analyses revealed functional integration among regions in the social brain, and suggest that the insula is positioned to interact with previously identified nodes important for social and affective behaviors.”

May combine two claims.

Exposure to a stressed conspecific evokes OT release within the insular cortex which, via modulation of insular cortex output neuron excitability, orchestrates species-specific, age-dependent approach or avoidance behaviors via modulation of the social decision-making network (SDMN).

“exposure to a stressed conspecific evokes OT release within the insular cortex which, via modulation of insular cortex output neuron excitability, orchestrates species-specific, age-dependent approach or avoidance behaviors via modulation of the social decision-making network (SDMN).”

May combine two claims.

In the insular cortex, optogenetic silencing, blockade of the OTR, and inhibition of PKC each interfered with experimental rat responses to stressed conspecifics.

“In the insular cortex, optogenetic silencing, blockade of the OTR, and inhibition of PKC each interfered with experimental rat responses to stressed conspecifics”

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

22kHz USVs are emitted as alarm signals, and frequency modulating USVs are thought to convey positive affect.

“22kHz USVs are emitted as alarm signals, and frequency modulating USVs are thought to convey positive affect39.”

• cites [39] Burgdorf, J. et al. Ultrasonic vocalizations of rats (Rattus norvegicus) during mating, play, and aggression: behavioral concomitants, relationship to reward, and self-administration of playback. J. Comp. Psychol. 122, 357–367 (2008). sourceNot checked yet

  Not checked in the free run.

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

In human social behavior, features of the target stimulus, including age, are critical determinants of whether or not someone will approach, help or avoid another in distress.

“In human social behavior, features of the target stimulus, including age, are critical determinants of whether or not someone will approach, help or avoid another in distress27”

• cites [27] Staub, E. Helping a distressed person: social, personality, and stimulus determinants. Adv. Exp. Soc. Psychol.. 7, 293–341 (1974). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Emotion recognition and empathic deficits occur after insular cortex lesions.

“emotion recognition and empathic deficits occur after insular cortex lesions3–5”

• cites [3] Gu, X. et al. Anterior insular cortex is necessary for empathetic pain perception. Brain 135, 2726–2735 (2012). sourceNot checked yet

  Not checked in the free run.

• cites [4] Ibañez, A., Gleichgerrcht, E. & Manes, F. Clinical effects of insular damage in humans. Brain Struct. Funct. 214, 397–410 (2010). sourceNot checked yet

  Not checked in the free run.

• cites [5] Leigh, R. et al. Acute lesions that impair affective empathy. Brain 136, 2539–2549 (2013). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Like social behavior itself, the effects of OT administration on social decision-making are sensitive to situational and interpersonal factors, with OT sometimes producing prosocial effects and at other times anti-social effects.

“Like social behavior itself, the effects of OT administration on social decision-making are sensitive to situational and interpersonal factors, with OT sometimes producing prosocial effects and at other times anti-social effects46”

• cites [46] Bartz, J. A., Zaki, J., Bolger, N. & Ochsner, K. N. Social effects of oxytocin in humans: context and person matter. Trends Cogn. Sci. 15, 301–309 (2011).Not checked yet

  Not checked in the free run.

May combine two claims.

The insular cortex receives input from oxytocin (OT)-containing axons from the paraventricular nucleus, and OT receptor (OTR) binding is enriched in the insular cortex.

“The insular cortex also receives input from oxytocin (OT)-containing axons from the paraventricular nucleus17, and OT receptor (OTR) binding is enriched in the insular cortex18.”

• cites [17] Knobloch, H. S. et al. Evoked axonal oxytocin release in the central amygdala attenuates fear response. Neuron 73, 553–566 (2012). sourceNot checked yet

  Not checked in the free run.

• cites [18] Dumais, K. M., Bredewold, R., Mayer, T. E. & Veenema, A. H. Sex differences in oxytocin receptor binding in forebrain regions: correlations with social interest in brain region- and sex- specific ways. Horm. Behav. 64, 693–701 (2013). sourceNot checked yet

  Not checked in the free run.

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

The decision to avoid a stressed adult may be adaptive as the stressed conspecific may be perceived as a social danger cue.

“The decision to avoid a stressed adult may be adaptive as the stressed conspecific may be perceived as a social danger cue.”

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

There are examples of prosocial responses to stressed conspecifics in both voles and mice.

“and there are examples of prosocial responses to stressed conspecifics in both voles21 and mice41.”

• cites [21] Burkett, J. P. et al. Oxytocin-dependent consolation behavior in rodents. Science 351, 375–378 (2016). sourceNot checked yet

  Not checked in the free run.

• cites [41] Langford, D. J. et al. Social approach to pain in laboratory mice. Soc. Neurosci. 5, 163–170 (2010). sourceNot checked yet

  Not checked in the free run.

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

Abnormalities in emotion recognition are central to several psychiatric conditions including ASD, in which OTR polymorphisms and insular cortex hypofunction are correlates of symptom severity.

“Abnormalities in emotion recognition are central to several psychiatric conditions including ASD47,48, in which OTR polymorphisms49,50 and insular cortex hypofunction8,9 are correlates of symptom severity.”

• cites [47] Decety, J. & Moriguchi, Y. The empathic brain and its dysfunction in psychiatric populations: implications for intervention across different clinical conditions. Biopsychosoc. Med.. 1, 22 (2007). sourceNot checked yet

  Not checked in the free run.

• cites [48] Thioux, M. & Keysers, C. Empathy: shared circuits and their dysfunctions. Dialog-. Clin. Neurosci. 12, 546–552 (2010). sourceNot checked yet

  Not checked in the free run.

• cites [49] LoParo, D. & Waldman, I. D. The oxytocin receptor gene (OXTR) is associated with autism spectrum disorder: a meta-analysis. Mol. Psychiatry 20, 640–646 (2015). sourceNot checked yet

  Not checked in the free run.

• cites [50] Skuse, D. H. et al. Common polymorphism in the oxytocin receptor gene (OXTR) is associated with human social recognition skills. Proc. Natl. Acad. Sci. USA 111, 1987–1992 (2014). sourceNot checked yet

  Not checked in the free run.

• cites [8] Morita, T. et al. Emotional responses associated with self-face processing in individuals with autism spectrum disorders: an fMRI study. Soc. Neurosci. 7, 223–239 (2012). sourceNot checked yet

  Not checked in the free run.

• cites [9] Odriozola, P. et al. Insula response and connectivity during social and non-social attention in children with autism. Soc. Cogn. Affect. Neurosci. 11, 433–444 (2016). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

We report a set of in vivo and in vitro studies that implicate insular cortex activity and modulation by OT as necessary and sufficient for modulating social affective behaviors in rat.

“We report a set of in vivo and in vitro studies that implicate insular cortex activity and modulation by OT as necessary and sufficient for modulating social affective behaviors in rat.”

May combine two claims.

Prosocial effects are evident in pair-bonded voles and familiar female mice, but not between strangers or in male mice; this may also generalize to rats.

“First, prosocial effects are evident in pair-bonded voles and familiar female mice, but not between strangers or in male mice; this may also generalize to rats42 (but see25).”

• cites [42] Ben-Ami Bartal, I., Rodgers, D. A., Bernardez Sarria, M. S., Decety, J. & Mason, P. Pro-social behavior in rats is modulated by social experience. eLife 3, e01385 (2014). sourceNot checked yet

  Not checked in the free run.

• cites [25] Silberberg, A. et al. Desire for social contact, not empathy, may explain “rescue” behavior in rats. Anim. Cogn. 17, 609–618 (2014). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

OT may bias the output of insula efferents to the ventral striatum or prefrontal cortex to promote interaction with stressed juveniles, or to the basolateral amygdala to promote avoidance in response to stressed adults.

“Thus, in concert with sensory cues that identify the age of the conspecific, OT may bias the output of insula efferents to the ventral striatum or prefrontal cortex to promote interaction with stressed juveniles, or to the basolateral amygdala to promote avoidance in response to stressed adults”

May combine two claims.

Insular cortex is interconnected with the SDMN; this connectivity includes inputs from the hypothalamus and ventral tegmental area, reciprocal connections with the amygdala and outputs to periaqueductal gray, bed nucleus of the stria terminalis, nucleus accumbens, and medial prefrontal cortex.

“Importantly, insular cortex is interconnected with the SDMN; this connectivity includes inputs from the hypothalamus11 and ventral tegmental area12, reciprocal connections with the amygdala13 and outputs to periaqueductal gray14, bed nucleus of the stria terminalis, nucleus accumbens15, and medial prefrontal cortex16.”

• cites [11] Allen, G. V. & Cechetto, D. F. Functional and anatomical organization of cardiovascular pressor and depressor sites in the lateral hypothalamic area. II. Ascending projections. J. Comp. Neurol. 330, 421–438 (1993). sourceNot checked yet

  Not checked in the free run.

• cites [12] Ohara, P. T. et al. Dopaminergic input to GABAergic neurons in the rostral agranular insular cortex of the rat. J. Neurocytol. 32, 131–141 (2003). sourceNot checked yet

  Not checked in the free run.

• cites [13] Shi, C. J. & Cassell, M. D. Cortical, thalamic, and amygdaloid connections of the anterior and posterior insular cortices. J. Comp. Neurol. 399, 440–468 (1998). sourceNot checked yet

  Not checked in the free run.

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

Recordings were included only if series resistance remained less than 30 MΩ with less than 10% change from baseline throughout the experiment.

“recordings were only included if the series resistance remained less than 30 MΩ with less than 10% change from baseline throughout the experiment.”

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

Translational rodent models that capture aspects of social affect include emotion contagion and social buffering.

“Translational rodent models that capture aspects of social affect include emotion contagion and social buffering24.”

• cites [24] de Waal, F. B. M. & Preston, S. D. Mammalian empathy: behavioural manifestations and neural basis. Nat. Rev. Neurosci. 18, 498–509 (2017). sourceNot checked yet

  Not checked in the free run.

Classified as probably not a substantial claim.

Rats with occluded injectors or cannula located outside the insular cortex were excluded from all analyses.

“Rats with occluded injectors or having cannula located outside of the insular cortex were excluded from all analyses (See Supplementary Fig. 6).”

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

Optogenetic transductions were evaluated by fluorescence microscopy and in vitro whole cell recordings.

“Optogenetic transductions were evaluated by fluorescence microscopy and in vitro whole cell recordings (Supplementary Fig. 5).”

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

In all behavioral experiments, rats were randomly assigned to treatments.

“In all behavioral experiments, rats were randomly assigned to treatments.”

May combine two claims.

OT modulates social behaviors across species, including empathic cognition in both humans and rodents.

“OT modulates social behaviors across species19, including empathic cognition in both humans20 and rodents21.”

• cites [19] Donaldson, Z. R. & Young, L. J. Oxytocin, vasopressin, and the neurogenetics of sociality. Science 322, 900–904 (2008). sourceNot checked yet

  Not checked in the free run.

• cites [20] Shamay-Tsoory, S. G. & Abu-Akel, A. The social salience hypothesis of oxytocin. Biol. Psychiatry 79, 194–202 (2016). sourceNot checked yet

  Not checked in the free run.

• cites [21] Burkett, J. P. et al. Oxytocin-dependent consolation behavior in rodents. Science 351, 375–378 (2016). sourceNot checked yet

  Not checked in the free run.

Classified as probably not a substantial claim.

V1a receptor mRNA has been reported throughout cortex.

“V1a receptor mRNA has been reported throughout cortex60”

• cites [60] Dumais, K. M. & Veenema, A. H. Vasopressin and oxytocin receptor systems in the brain: Sex differences and sex-specific regulation of social behavior. Front. Neuroendocrinol. 40, 1–23 (2016). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

OT relieves social deficits observed in ASD and augments insular activity and connectivity, suggesting a mechanistic relationship between OT, insula and social cognition.

“OT relieves social deficits observed in ASD and augments insular activity and connectivity22,23, suggesting a mechanistic relationship between OT, insula and social cognition.”

• cites [22] Aoki, Y. et al. Oxytocin improves behavioural and neural deficits in inferring others’ social emotions in autism. Brain 137, 3073–3086 (2014). sourceNot checked yet

  Not checked in the free run.

• cites [23] Scheele, D. et al. An oxytocin-induced facilitation of neural and emotional responses to social touch correlates inversely with autism traits. Neuropsychopharmacology 39, 2078–2085 (2014). sourceNot checked yet

  Not checked in the free run.

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

Only neurons with pyramidal morphology and soma in deep layers of insular cortex were included for analysis.

“Only neurons with a pyramidal morphology and soma in deep layers of insular cortex were included for analysis.”

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

Sample sizes were initially determined based on prior work using social interaction and intrinsic physiology, and no formal statistical methods were used to pre-determine sample sizes.

“Sample sizes were initially determined based on prior work using social interaction54,65 and intrinsic physiology62, no formal statistical methods were used to pre-determine sample sizes.”

• cites [54] Manning, M. et al. Oxytocin and vasopressin agonists and antagonists as research tools and potential therapeutics. J. Neuroendocrinol. 24, 609–628 (2012).Not checked yet

  Not checked in the free run.

• cites [65] Vetere, G. et al. Chemogenetic interrogation of a brain-wide fear memory network in mice. Neuron 94, 363–374.e4 (2017). sourceNot checked yet

  Not checked in the free run.

• cites [62] Turrigiano, G. G., Leslie, K. R., Desai, N. S., Rutherford, L. C. & Nelson, S. B. Activity-dependent scaling of quantal amplitude in neocortical neurons. Nature 391, 892–896 (1998). sourceNot checked yet

  Not checked in the free run.

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

Inter-rater reliability was determined by a second observer completely blind to experimental manipulations and hypotheses.

“Digital video recordings were made of each test for later determination of inter-rater-reliability by a second observer completely blind to the experimental manipulations and hypotheses.”

May combine two claims.

The observed change in mEPSC amplitude likely reflects postsynaptic modulation of AMPA receptor-mediated currents, not an effect of OT on spontaneous circuit activity per se.

“The change in mEPSC amplitude observed likely reflects a postsynaptic modulation of AMPA receptor mediated currents63 but not an effect of OT on the spontaneous circuit activity per se.”

• cites [63] Gschwendt, M. et al. Inhibition of protein kinase C mu by various inhibitors. Differentiation from protein kinase c isoenzymes. FEBS Lett.. 392, 77–80 (1996). sourceNot checked yet

  Not checked in the free run.

• AssumptionThe authors state this as an assumption: All events recorded in the mEPSC analysis were miniature EPSCs.

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

In one-on-one social exploration tests, exploratory behaviors were timed by an observer blind to treatment.

“exploratory behaviors (sniffing, pinning, and allogrooming) initiated by the adult test subject were timed by an observer blind to treatment.”

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

Videos used for additional behavioral quantification were selected blind to SAP test social interaction results.

“Videos were selected blind to SAP test social interaction results.”

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

In one-on-one social exploration tests, test order was counterbalanced.

“test order was counterbalanced.”

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

Juvenile and adult stimuli rats were used for multiple tests but never more than once for the same adult test rat.

“Juvenile and adult stimuli rats were used for multiple tests but were never used more than once for the same adult test rat.”

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

The dose of 500 nM OT was selected after a pilot study using a range of doses from 50 nM to 1 μM, the largest dose reported.

“The dose of 500 nM was selected after a pilot study using a range of doses from 50nM to 1µM, the largest dose reported59.”

• cites [59] Szot, P., Bale, T. L. & Dorsa, D. M. Distribution of messenger RNA for the vasopressin V1a receptor in the CNS of male and female rats. Brain Res. Mol. Brain Res. 24, 1–10 (1994). sourceNot checked yet

  Not checked in the free run.

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

Main effects and interactions were deemed significant when p < 0.05 and all reported post hoc test p values are Sidak-adjusted to maintain an experiment-wise risk of type I errors at α = 0.05.

“Main effects and interactions were deemed significant when p < 0.05 and all reported post hoc test p values are Sidak-adjusted, to maintain an experiment-wise risk of type I errors at α = 0.05.”

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

Data were collected by observers blind to treatment.

“Data were collected by observers blind to treatment.”

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

Final data analyses were not performed blind to the conditions of the experiments.

“Final data analyses were not performed blind to the conditions of the experiments.”

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

Graph theoretical analysis has been applied to Fos datasets to characterize functional relationships in rodent neural circuits.

“Graph theoretical analysis has been applied to Fos datasets to characterize functional relationships in rodent neural circuits66–69.”

• cites [66] Wheeler, A. L. et al. Identification of a functional connectome for long-term fear memory in mice. PLOS Comput. Biol. 9, e1002853 (2013). sourceNot checked yet

  Not checked in the free run.

• cites [67] Tanimizu, T. et al. Functional connectivity of multiple brain regions required for the consolidation of social recognition memory. J. Neurosci. 37, 4103–4116 (2017). sourceNot checked yet

  Not checked in the free run.

• cites [68] Teles, M. C., Almeida, O., Lopes, J. S. & Oliveira, R. F. Social interactions elicit rapid shifts in functional connectivity in the social decision-making network of zebrafish. Proc. Biol. Sci.. 282, 20151099 (2015). sourceNot checked yet

  Not checked in the free run.

• cites [69] Newman, M. E. Modularity and community structure in networks. Proc. Natl. Acad. Sci. USA 103, 8577–8582 (2006). sourceNot checked yet

  Not checked in the free run.

Classified as probably not a substantial claim.

44 adult male rats were randomly assigned to one of four treatment conditions in a 2 by 2 (Stress by Age) design (n=11/group).

“44 adult male rats were habituated for 1h to the test arena and 24h later randomly assigned to one of 4 treatment conditions: Naive-Juvenile, Naive-Adult, Stressed-Juvenile or Stressed-Adult in a 2 by 2 (Stress by Age) design (n = 11/group).”

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

USV detection parameters were refined for each recording based on visual inspection by a trained observer blind to treatment condition.

“detection parameters were refined for each recording based on visual inspection by a trained observer who was blind to treatment condition.”

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

Participation coefficients were calculated to determine the degree to which nodes participated in multiple networks, based on arguments that this measure is the most appropriate way to identify hubs in correlation-based networks.

“Participation coefficients were calculated to determine the degree to which nodes participated in multiple networks, based on arguments that this measure is the most appropriate way to identify “hubs” in correlation-based networks32.”

• cites [32] Power, J. D., Schlaggar, B. L., Lessov-Schlaggar, C. N. & Petersen, S. E. Evidence for hubs in human functional brain networks. Neuron 79, 798–813 (2013). sourceNot checked yet

  Not checked in the free run.

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

In electrophysiology experiments, cells were treated according to a Latin square design to achieve even representation of cells in treatment groups from different donor rats.

“In electrophysiology experiments, cells were treated according to a Latin square design to achieve even representation of cells in treatment groups from different donor rats.”

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

The OT dose was increased to 1 μM in the fEPSP experiment because 500 nM did not reliably replicate the increase in fEPSPs when DMSO was added to the aCSF.

“The OT dose was increased to 1µM in the fEPSP experiment as 500nM did not reliably replicate the increase in fEPSPs when DMSO was added to the aCSF.”