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Cited by 95Regulation of Appetite and ObesityNeurotransmitter Receptor Influence on BehaviorStress Responses and Cortisol

Chronic unpredictable stress induces depression-related behaviors by suppressing AgRP neuron activity

Open access
Fang, Xing, Jiang, Shujun, Wang, Jiangong, Bai, Yu, Kim, Chung Sub, Blake, David, Weintraub, Neal L., Lei, Yun +1 more·2021·Molecular Psychiatry
DOI 10.1038/s41380-020-01004-xPMC PMC8272726
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Abstract

Previous studies have shown that AgRP neurons in the arcuate nucleus (ARC) respond to energy deficits and play a key role in the control of feeding behavior and metabolism. Here, we demonstrate that chronic unpredictable stress, an animal model of depression, decreases spontaneous firing rates, increases firing irregularity and alters the firing properties of AgRP neurons in both male and female mice. These changes are associated with enhanced inhibitory synaptic transmission and reduced intrinsic neuronal excitability. Chemogenetic inhibition of AgRP neurons increases susceptibility to subthreshold unpredictable stress. Conversely, chemogenetic activation of AgRP neurons completely reverses anhedonic and despair behaviors induced by chronic unpredictable stress. These results indicate that chronic stress induces maladaptive synaptic and intrinsic plasticity, leading to hypoactivity of AgRP neurons and subsequently causing behavioral changes. Our findings suggest that AgRP neurons in the ARC are a key component of neural circuitry involved in mediating depression-related behaviors and that increasing AgRP neuronal activity coule be a novel and effective treatment for depression.

AI snapshot

Written from the paper’s full text.

TL;DR

Chronic unpredictable stress suppresses AgRP neuron activity in male and female mice via increased inhibitory synaptic transmission and reduced intrinsic excitability, and this hypoactivity contributes to depression-related behaviors. Chemogenetic activation of AgRP neurons reverses stress-induced anhedonia and despair behavior.

Key findings

  • Chronic unpredictable stress reduced sucrose preference in male (P = 0.030) and female (P = 0.004) mice and increased forced swim test immobility in male (P = 0.037) and female (P = 0.012) mice, without altering locomotor activity.
  • AgRP neurons showed decreased spontaneous firing rates (male P = 0.033; female P = 0.045; total P = 0.003), hyperpolarized membrane potential, and increased firing irregularity after chronic stress.
  • Chronic stress increased inhibitory synaptic transmission onto AgRP neurons: spontaneous IPSC amplitude was increased (male P = 0.038; female P = 0.001; total P < 0.001), and miniature IPSC amplitude was increased (P = 0.012), while excitatory currents were unchanged.
  • Intrinsic AgRP neuron excitability was reduced (firing rate P = 0.011), silent neurons increased from 5% (1/24) to 23% (6/26), and afterhyperpolarization amplitude increased (P < 0.001). Chemogenetic inhibition increased stress susceptibility, whereas activation reversed anhedonia and despair behavior.

Methods

  • Male and female C57BL/6J and AgRP-ires-Cre;tdTomato mice underwent 10 days of chronic unpredictable stress (stressors included restraint, tail pinch, constant light, wet bedding/cage tilt, foot shocks, elevated platform, and social isolation), followed by sucrose preference, female urine sniffing, forced swim, and open field tests.
  • Whole-cell patch-clamp recordings were made from tdTomato-labeled AgRP neurons in the arcuate nucleus to measure spontaneous and miniature EPSCs/IPSCs, intrinsic firing, and action potential properties.
  • Chemogenetic manipulation used ARC injections of AAV-DIO-hM4Di-mCherry, AAV-DIO-hM3Dq-mCherry, or AAV-DIO-mCherry in AgRP-ires-Cre mice, with CNO (0.3 mg/kg, i.p.) before behavioral testing; c-Fos and AgRP immunohistochemistry verified activation and cell survival.

Limitations

  • The paper states that it is not clear which downstream targets of AgRP neurons contribute to depression-related behaviors and that future studies are needed to examine VTA and BNST involvement.
  • Evident from the text: miniature EPSC/IPSC recordings and the female urine sniffing test were performed only in male mice, limiting sex-specific conclusions for those measures.
  • Evident from the text: intrinsic excitability recordings came from a small number of mice per group (Ctrl: 24 neurons from 3 mice; CUS: 26 neurons from 3 mice), and all findings are from mouse models rather than human data.

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

Integrity signals

Registry

Crossref · 10.1038/s41380-020-01004-x

Cites

94 of 94 references verified0 citation errors · 0 not checkable

Statistics

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

Claims the paper makes

0 of 107 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    Chronic unpredictable stress causes a right shift in the cumulative frequency distribution of interspike intervals of AgRP neurons in both male and female mice.

    “A right shift of the cumulative frequency distribution of ISIs of different lengths”
    Quote in the paperSubstantial claim
  • Findingown data

    Chemogenetic activation of AgRP neurons completely reverses anhedonic and despair behaviors induced by chronic unpredictable stress.

    “Conversely, chemogenetic activation of AgRP neurons completely reverses anhedonic and despair behaviors induced by chronic unpredictable stress.”
    Quote in the paperSubstantial claim
  • Findingown data

    Chronic unpredictable stress reduces time spent sniffing female urine in male mice compared with non-stressed controls.

    “Male mice exposed to chronic unpredictable stress spent less time sniffing female urine when compared with non-stressed controls”
    Quote in the paperSubstantial claim
  • Findingown data

    Chronic unpredictable stress increases immobility in the forced swim test in both male and female mice.

    “chronically stressed mice of both sexes exhibited increased immobility in the forced swim test”
    Quote in the paperSubstantial claim
  • Findingown data

    Chemogenetic inhibition of AgRP neurons increases susceptibility to subthreshold unpredictable stress.

    “Chemogenetic inhibition of AgRP neurons increases susceptibility to subthreshold unpredictable stress.”
    Quote in the paperSubstantial claim
  • May combine two claims. 25
  • The quote only partly states this claim. 4
  • Classified as probably not a substantial claim. 51

  • May combine two claims.

    Findingown data

    The changes in AgRP neuron firing induced by chronic unpredictable stress are associated with enhanced inhibitory synaptic transmission and reduced intrinsic neuronal excitability.

    “These changes are associated with enhanced inhibitory synaptic transmission and reduced intrinsic neuronal excitability.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    There was a negative correlation between intrinsic firing rates and coefficients of variation of ISIs under control (r = −0.669, P < 0.001) and chronic stress (r = −0.604, P = 0.001) conditions, with steeper slopes in chronically stressed mice than control mice.

    “There was a negative correlation between intrinsic firing rates and coefficients of variation of ISIs under control and chronic stress conditions (Fig. 3e; Ctrl: r = −0.669, P < 0.001; CUS: r = −0.604, P = 0.001) with steeper slopes in chronically stressed mice than control mice (Fig. 3e).”
    Quote in the paperNot a substantial claim

  • The quote only partly states this claim.

    Findingown data

    The intrinsic firing frequency of AgRP neurons was decreased in mice of both sexes after 10 days of unpredictable stress (P = 0.011).

    “The intrinsic firing frequency was decreased (Fig. 3b, c, Mann Whitney test, P = 0.011)”
    • Also stated in: § Chronic unpredictable stress alters the intrinsic firing rate, patterns, and shape of AgRP neurons · boosted
      “The intrinsic activity of AgRP neurons was clearly reduced in mice of both sexes after exposure to 10 days of unpredictable stress (Fig. 3a, b).”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    Induction of c-Fos immunoreactivity was observed in the ARC of hM3Dq-expressing mice 2 h after CNO injection (0.3 mg/kg, i.p.).

    “Induction of c-Fos immunoreactivity was observed in the ARC of hM3Dq-expressing mice (Fig. 5c), indicating CNO-induced activation of AgRP neurons.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Dampened AgRP neuron activity was observed only after a full course of chronic exposure (10 days) to unpredictable stress, but not after short-term subthreshold exposure (3 days).

    “The dampened AgRP neuron activity was only observed after a full course of chronic exposure (i.e., 10 days) to unpredictable stress, but not after short-term subthreshold exposure (i.e., 3 days).”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    Immobility time in the forced swim test was decreased by stimulation of AgRP neurons in female mice (P = 0.019).

    “Immobility time in the forced swim test was decreased by stimulation of AgRP neurons (Fig. 5e2, one-way ANOVA, F(2,36) = 4.458, P = 0.019)”
    Quote in the paperSubstantial claim

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

    Findingown data

    AgRP neurons from chronically stressed mice exhibited unaltered action potential rise time (P = 0.073).

    “AgRP neurons from chronically stressed mice exhibited unaltered rise time (Mann Whitney test, P = 0.073)”
    Quote in the paperNot a substantial claim

  • The quote only partly states this claim.

    Findingown data

    There is a correlation between the degree of firing irregularity and the firing rate for individual AgRP neurons.

    “AgRP neurons from chronically stressed mice fired more slowly and irregularly, with a correlation between the degree of firing irregularity and the firing rate for individual neurons.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    The amplitude but not the frequency of spontaneous IPSCs was increased by chronic unpredictable stress in AgRP neurons.

    “The amplitude but not the frequency of spontaneous IPSCs was increased by chronic unpredictable stress, suggesting enhanced postsynaptic responsiveness.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Chronic unpredictable stress increased inhibitory synaptic transmission without changing excitatory synaptic transmission in AgRP neurons.

    “Recordings of spontaneous excitatory and inhibitory synaptic currents in AgRP neurons demonstrated that chronic unpredictable stress increased inhibitory synaptic transmission without changing excitatory synaptic transmission.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    AgRP neurons are found exclusively in the arcuate nucleus.

    “Neurons expressing agouti-related protein (AgRP) are found exclusively in the ARC [20]”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [20] Bagnol D, Lu XY, Kaelin CB, Day HE, Ollmann M, Gantz I, et al. Anatomy of an endogenous antagonist: relationship between Agouti-related protein and proopiomelanocortin in brain. J Neurosci. 1999;19:RC26. source

      Not checked in the free run.

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

    Premisecitation

    AgRP neurons are inhibited by overfeeding and satiety signals.

    “and inhibited by overfeeding and satiety signals [23–25].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [23] Mandelblat-Cerf Y, Ramesh RN, Burgess CR, Patella P, Yang ZF, Lowell BB et al. Arcuate hypothalamic AgRP and putative POMC neurons show opposite changes in spiking across multiple timescales. Elife. 2015;4:e07122. source

      Not checked in the free run.

    • cites [24] Baver SB, Hope K, Guyot S, Bjorbaek C, Kaczorowski C, O’Connell KM. Leptin modulates the intrinsic excitability of AgRP/NPY neurons in the arcuate nucleus of the hypothalamus. J Neurosci. 2014;34:5486–96. source

      Not checked in the free run.

    • cites [25] Chen Y, Lin YC, Kuo TW, Knight ZA. Sensory detection of food rapidly modulates arcuate feeding circuits. Cell. 2015;160:829–41. source

      Not checked in the free run.

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

    Premisecitation

    AgRP neurons are stimulated by hunger signals.

    “and they are stimulated by hunger signals [21, 22]”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [21] Morton GJ, Cummings DE, Baskin DG, Barsh GS, Schwartz MW. Central nervous system control of food intake and body weight. Nature. 2006;443:289–95. source

      Not checked in the free run.

    • cites [22] Hahn TM, Breininger JF, Baskin DG, Schwartz MW. Coexpression of Agrp and NPY in fasting-activated hypothalamic neurons. Nat Neurosci. 1998;1:271–2. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Chronic stress induces maladaptive synaptic and intrinsic plasticity, leading to hypoactivity of AgRP neurons and subsequently causing behavioral changes.

    “These results indicate that chronic stress induces maladaptive synaptic and intrinsic plasticity, leading to hypoactivity of AgRP neurons and subsequently causing behavioral changes.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    AgRP neurons in the arcuate nucleus respond to energy deficits and play a key role in the control of feeding behavior and metabolism.

    “Previous studies have shown that AgRP neurons in the arcuate nucleus (ARC) respond to energy deficits and play a key role in the control of feeding behavior and metabolism.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    AgRP neurons are involved in feeding and food reward processing.

    “These neurons are well known to be involved in feeding and food reward processing [18, 19].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [18] Cone RD. Anatomy and regulation of the central melanocortin system. Nat Neurosci. 2005;8:571–8. source

      Not checked in the free run.

    • cites [19] Gao Q, Horvath TL. Neurobiology of feeding and energy expenditure. Annu Rev Neurosci. 2007;30:367–98. source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisecitation

    Neurons in the arcuate nucleus of the hypothalamus are activated by emotional stimuli.

    “We and others have previously shown that neurons in the arcuate nucleus (ARC) of the hypothalamus are activated by emotional stimuli [16, 17].”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [16] Liu J, Garza JC, Truong HV, Henschel J, Zhang W, Lu XY. The melanocortinergic pathway is rapidly recruited by emotional stress and contributes to stress-induced anorexia and anxiety-like behavior. Endocrinology. 2007;148:5531–40. source

      Not checked in the free run.

    • cites [17] Chagra SL, Zavala JK, Hall MV, Gosselink KL. Acute and repeated restraint differentially activate orexigenic pathways in the rat hypothalamus. Regul Pept. 2011;167:70–8. source

      Not checked in the free run.

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    Study design and conductnone given

    Behaviors were scored by experimenters blinded to the treatments.

    “Behaviors were scored by the experimenters who were blinded to the treatments.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    AgRP neurons can respond rapidly to acute emotional stress.

    “We have previously demonstrated that AgRP neurons can respond rapidly to acute emotional stress [16].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [16] Liu J, Garza JC, Truong HV, Henschel J, Zhang W, Lu XY. The melanocortinergic pathway is rapidly recruited by emotional stress and contributes to stress-induced anorexia and anxiety-like behavior. Endocrinology. 2007;148:5531–40. source

      Not checked in the free run.

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    Premisecitation

    Anhedonia is a robust predictor of suicidality.

    “It has also been considered as a robust predictor of suicidality [10, 11].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [10] Harrington R. Depression, suicide and deliberate self-harm in adolescence. Br Med Bull. 2001;57:47–60. source

      Not checked in the free run.

    • cites [11] Cipriani A, Barbui C, Geddes JR. Suicide, depression, and antidepressants. BMJ. 2005;330:373–4. source

      Not checked in the free run.

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

    Premisenone given

    The chronic unpredictable/mild/variable stress model allows comparison of the effects of stress between males and females.

    “One major advantage of the chronic unpredictable/mild/variable stress model is that it allows us to compare the effects of stress between males and females.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    Inhibition of AgRP neurons suppresses feeding.

    “Conversely, inhibition of these neurons suppresses feeding [27].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [27] Krashes MJ, Koda S, Ye C, Rogan SC, Adams AC, Cusher DS, et al. Rapid, reversible activation of AgRP neurons drives feeding behavior in mice. J Clin Investig. 2011;121:1424–8. source

      Not checked in the free run.

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

    Premisecitation

    Ablation of AgRP neurons enhances isolation stress-induced anorexia and palatability-driven feeding.

    “Paradoxically, ablation of AgRP neurons has been reported to enhance isolation stress-induced anorexia and palatability-driven feeding [28].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [28] Denis RGP, Joly-Amado A, Webber E, Langlet F, Schaeffer M, Padilla SL, et al. Palatability can drive feeding independent of AgRP neurons. Cell Metab. 2015;22:646–57. source

      Not checked in the free run.

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

    Premisecitation

    Anhedonia typically does not respond to first-line pharmacotherapies.

    “Anhedonia typically does not respond to first-line pharmacotherapies [5, 7, 8]”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [5] Treadway MT, Zald DH. Reconsidering anhedonia in depression: lessons from translational neuroscience. Neurosci Biobehav Rev. 2011;35:537–55. source

      Not checked in the free run.

    • cites [7] McMakin DL, Olino TM, Porta G, Dietz LJ, Emslie G, Clarke G, et al. Anhedonia predicts poorer recovery among youth with selective serotonin reuptake inhibitor treatment-resistant depression. J Am Acad Child Adolesc Psychiatry. 2012;51:404–11. source

      Not checked in the free run.

    • cites [8] Tran P, Skolnick P, Czobor P, Huang NY, Bradshaw M, McKinney A, et al. Efficacy and tolerability of the novel triple reuptake inhibitor amitifadine in the treatment of patients with major depressive disorder: a randomized, double-blind, placebo-controlled trial. J Psychiatr Res. 2012;46:64–71. source

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    Premisecitation

    Anhedonia is one of the two core symptoms required for the diagnosis of major depression.

    “anhedonia is one of the two core symptoms required for the diagnosis [2–4].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [2] Gorwood P. Neurobiological mechanisms of anhedonia. Dialog Clin Neurosci. 2008;10:291–9. source

      Not checked in the free run.

    • cites [3] Rizvi SJ, Pizzagalli DA, Sproule BA, Kennedy SH. Assessing anhedonia in depression: potentials and pitfalls. Neurosci Biobehav Rev. 2016;65:21–35. source

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    • cites [4] Association AP. Diagnostic and statistical manual of mental disorders. 5th ed. Washington, DC: American Psychiatric Press; 2013.

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    Study design and conductnone given

    All animal procedures were approved by the Institutional Animal Care and use Committees of the University of Texas Health Science Center at San Antonio and Augusta University.

    “all animal procedures were approved by the Institutional Animal Care and use Committees of the University of Texas Health Science Center at San Antonio and Augusta University.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Anhedonia is usually the last symptom to resolve in depression.

    “and is usually the last symptom to resolve in depression [9].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [9] Rubin DH. Joy returns last: anhedonia and treatment resistance in depressed adolescents. J Am Acad Child Adolesc Psychiatry. 2012;51:353–5. source

      Not checked in the free run.

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    Premisecitation

    A recent study failed to show a change in AgRP neuron activity after repeated restraint stress.

    “However, a recent study failed to show a change in AgRP neuron activity after repeated restraint stress [29].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [29] Qu N, He Y, Wang C, Xu P, Yang Y, Cai X, et al. A POMC-originated circuit regulates stress-induced hypophagia, depression, and anhedonia. Mol Psychiatry. 2020;25:1006–21. source

      Not checked in the free run.

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

    Premisecitation

    Optogenetic and chemogenetic activation of AgRP neurons can evoke feeding in fed mice and promote motivationally, food-seeking behavior.

    “Optogenetic and chemogenetic activation of AgRP neurons can evoke feeding in fed mice and promote motivationally, food-seeking behavior [26, 27].”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [26] Aponte Y, Atasoy D, Sternson SM. AGRP neurons are sufficient to orchestrate feeding behavior rapidly and without training. Nat Neurosci. 2011;14:351–5. source

      Not checked in the free run.

    • cites [27] Krashes MJ, Koda S, Ye C, Rogan SC, Adams AC, Cusher DS, et al. Rapid, reversible activation of AgRP neurons drives feeding behavior in mice. J Clin Investig. 2011;121:1424–8. source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisecitation

    Chronic stress increases the risk of developing depression in humans.

    “As chronic stress increases the risk of developing depression in humans [30]”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [30] Kessler RC. The effects of stressful life events on depression. Annu Rev Psychol. 1997;48:191–214. source

      Not checked in the free run.

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

    Study design and conductnone given

    Injection sites were verified by examining mCherry fluorescence, and animals with mis-injections were excluded from statistical analysis.

    “The injection sites were verified by examining mCherry fluorescence in each animal at the end of the experiments. The animals with mis-injections were excluded from statistical analysis.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    The induction of anhedonia has been the primary focus in the chronic unpredictable/mild/variable stress model.

    “The induction of anhedonia has been the primary focus in this model”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [38] Willner P. Reliability of the chronic mild stress model of depression: a user survey. Neurobiol Stress. 2017;6:68–77. source

      Not checked in the free run.

    • cites [40] Parker KJ, Schatzberg AF, Lyons DM. Neuroendocrine aspects of hypercortisolism in major depression. Horm Behav. 2003;43:60–6. source

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

    Premisecitation

    Chronic unpredictable/mild/variable stress and chronic social defeat stress are the two most commonly used animal models of depression.

    “Chronic unpredictable/mild/variable stress and chronic social defeat stress are the two most commonly used animal models of depression [31, 34–38].”
    Quote in the paperNot a substantial claim0 of 6 sources checked
    • cites [31] Krishnan V, Nestler EJ. Animal models of depression: molecular perspectives. Curr Top Behav Neurosci. 2011;7:121–47. source

      Not checked in the free run.

    • cites [34] Lu XY, Kim CS, Frazer A, Zhang W. Leptin: a potential novel antidepressant. Proc Natl Acad Sci USA. 2006;103:1593–8. source

      Not checked in the free run.

    • cites [35] Liu J, Perez SM, Zhang W, Lodge DJ, Lu XY. Selective deletion of the leptin receptor in dopamine neurons produces anxiogenic-like behavior and increases dopaminergic activity in amygdala. Mol Psychiatry. 2011;16:1024–38. source

      Not checked in the free run.

    • cites [36] Lei Y, Wang J, Wang D, Li C, Liu B, Fang X, et al. SIRT1 in forebrain excitatory neurons produces sexually dimorphic effects on depression-related behaviors and modulates neuronal excitability and synaptic transmission in the medial prefrontal cortex. Mol Psychiatry. 2020;25:1094–111. source

      Not checked in the free run.

    • cites [37] Yu T, Guo M, Garza J, Rendon S, Sun XL, Zhang W, et al. Cognitive and neural correlates of depression-like behaviour in socially defeated mice: an animal model of depression with cognitive dysfunction. Int J Neuropsychopharmacol. 2011;14:303–17. source

      Not checked in the free run.

    • cites [38] Willner P. Reliability of the chronic mild stress model of depression: a user survey. Neurobiol Stress. 2017;6:68–77. source

      Not checked in the free run.

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

    Premisecitation

    Hypercortisolism is the most consistent biological abnormality in major depression.

    “has been the most consistent biological abnormality in this disorder [38, 40].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [38] Willner P. Reliability of the chronic mild stress model of depression: a user survey. Neurobiol Stress. 2017;6:68–77. source

      Not checked in the free run.

    • cites [40] Parker KJ, Schatzberg AF, Lyons DM. Neuroendocrine aspects of hypercortisolism in major depression. Horm Behav. 2003;43:60–6. source

      Not checked in the free run.

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    Premisecitation

    AgRP neurons receive both excitatory and inhibitory synaptic inputs.

    “AgRP neurons receive both excitatory and inhibitory synaptic inputs [62].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [62] Wang D, He X, Zhao Z, Feng Q, Lin R, Sun Y, et al. Whole-brain mapping of the direct inputs and axonal projections of POMC and AgRP neurons. Front Neuroanat. 2015;9:40. source

      Not checked in the free run.

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    Premisecitation

    The chronic unpredictable stress paradigm models unanticipated stressful experiences that occur in humans and has been widely used as a valid animal model of depression.

    “This paradigm models unanticipated stressful experiences that occur in humans, and thus has been widely used as a valid animal model of depression [34, 36, 52].”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [34] Lu XY, Kim CS, Frazer A, Zhang W. Leptin: a potential novel antidepressant. Proc Natl Acad Sci USA. 2006;103:1593–8. source

      Not checked in the free run.

    • cites [36] Lei Y, Wang J, Wang D, Li C, Liu B, Fang X, et al. SIRT1 in forebrain excitatory neurons produces sexually dimorphic effects on depression-related behaviors and modulates neuronal excitability and synaptic transmission in the medial prefrontal cortex. Mol Psychiatry. 2020;25:1094–111. source

      Not checked in the free run.

    • cites [52] Garza JC, Guo M, Zhang W, Lu XY. Leptin restores adult hippocampal neurogenesis in a chronic unpredictable stress model of depression and reverses glucocorticoid-induced inhibition of GSK-3beta/beta-catenin signaling. Mol Psychiatry. 2012;17:790–808. source

      Not checked in the free run.

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

    Premisecitation

    AgRP neurons in the ARC of the hypothalamus contribute to reward-driven feeding and food-seeking behavior.

    “A number of studies have shown that AgRP neurons in the ARC of the hypothalamus contribute to reward-driven feeding [28] and food-seeking behavior [26, 27].”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [28] Denis RGP, Joly-Amado A, Webber E, Langlet F, Schaeffer M, Padilla SL, et al. Palatability can drive feeding independent of AgRP neurons. Cell Metab. 2015;22:646–57. source

      Not checked in the free run.

    • cites [26] Aponte Y, Atasoy D, Sternson SM. AGRP neurons are sufficient to orchestrate feeding behavior rapidly and without training. Nat Neurosci. 2011;14:351–5. source

      Not checked in the free run.

    • cites [27] Krashes MJ, Koda S, Ye C, Rogan SC, Adams AC, Cusher DS, et al. Rapid, reversible activation of AgRP neurons drives feeding behavior in mice. J Clin Investig. 2011;121:1424–8. source

      Not checked in the free run.

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

    Premisecitation

    Research efforts aimed at understanding the neural circuits underlying depression have been mainly devoted to the prefrontal cortex, hippocampus, and mesolimbic structures.

    “Research efforts aimed at understanding the neural circuits underlying depression have been mainly devoted to the prefrontal cortex, hippocampus, and mesolimbic structures [12, 55, 56].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [12] Koenigs M, Grafman J. The functional neuroanatomy of depression: distinct roles for ventromedial and dorsolateral prefrontal cortex. Behav Brain Res. 2009;201:239–43. source

      Not checked in the free run.

    • cites [55] Duman RS, Monteggia LM. A neurotrophic model for stress-related mood disorders. Biol Psychiatry. 2006;59:1116–27. source

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    • cites [56] Nestler EJ, Carlezon WA. The mesolimbic dopamine reward circuit in depression. Biol Psychiatry. 2006;59:1151–9. source

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  • May combine two claims.

    Interpretationown data

    The impact of stress on AgRP neuron firing seems to depend not only on the type, intensity and duration of stress but also on the patterns of stress exposure and the predictability and controllability of stress.

    “The impact of stress on AgRP neuron firing seems to be dependent not only upon the type, intensity, and duration of stress but also upon the patterns of stress exposure and the nature of predictability and controllability of stress.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Along with depressed mood, anhedonia is one of the two main diagnostic criteria for major depression.

    “Along with depressed mood, anhedonia is one of the two main diagnostic criteria for major depression [2].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [2] Gorwood P. Neurobiological mechanisms of anhedonia. Dialog Clin Neurosci. 2008;10:291–9. source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisecitation

    A recent study using repeated restraint stress in a predictable fashion failed to show any changes in AgRP neuron activity.

    “a recent study using repeated restraint stress in a predictable fashion failed to show any changes in AgRP neuron activity [29].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [29] Qu N, He Y, Wang C, Xu P, Yang Y, Cai X, et al. A POMC-originated circuit regulates stress-induced hypophagia, depression, and anhedonia. Mol Psychiatry. 2020;25:1006–21. source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisecitation

    Repeated exposure to a homotypic stressor in a predictable manner (e.g., at the same time of day) has been used as a model of stress habituation, in which responses to a given stressor dampen after repeated presentations.

    “Exposure to a homotypic (same) stressor repeatedly in a predictable manner (e.g., at the same time of day) has been used as a model of stress habituation wherein responses to a given stressor dampen after repeated presentations [57].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [57] Herman JP. Neural control of chronic stress adaptation. Front Behav Neurosci. 2013;7:61. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    The stress-induced changes in AgRP neurons were associated with enhanced inhibitory synaptic transmission and reduced intrinsic neuronal excitability.

    “These changes were associated with enhanced inhibitory synaptic transmission and reduced intrinsic neuronal excitability.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Chronic stress induces maladaptive synaptic and intrinsic plasticity of AgRP neurons, which may contribute to the development of depression.

    “These data indicate for the first time that chronic stress induces maladaptive synaptic and intrinsic plasticity of AgRP neurons, which may contribute to the development of depression.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Regulation of spontaneous firing of AgRP neurons can be achieved by synaptic and intrinsic mechanisms.

    “Regulation of spontaneous firing of AgRP neurons can be achieved by synaptic and intrinsic mechanisms [54, 58–61].”
    Quote in the paperNot a substantial claim0 of 5 sources checked
    • cites [54] Takahashi KA, Cone RD. Fasting induces a large, leptin-dependent increase in the intrinsic action potential frequency of orexigenic arcuate nucleus neuropeptide Y/Agouti-related protein neurons. Endocrinology. 2005;146:1043–7. source

      Not checked in the free run.

    • cites [58] Zeltser LM, Seeley RJ, Tschop MH. Synaptic plasticity in neuronal circuits regulating energy balance. Nat Neurosci. 2012;15:1336–42. source

      Not checked in the free run.

    • cites [59] Dietrich MO, Horvath TL. Hypothalamic control of energy balance: insights into the role of synaptic plasticity. Trends Neurosci. 2013;36:65–73. source

      Not checked in the free run.

    • cites [60] Horvath TL. Synaptic plasticity in energy balance regulation. Obesity. 2006;14(Suppl 5):228S–233S. source

      Not checked in the free run.

    • cites [61] van den Top M, Lee K, Whyment AD, Blanks AM, Spanswick D. Orexigen-sensitive NPY/AgRP pacemaker neurons in the hypothalamic arcuate nucleus. Nat Neurosci. 2004;7:493–4. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    The chronic unpredictable stress paradigm effectively induces anhedonia along with other depression-related behaviors, including reduced sensitivity to sucrose reward, decreased sex-related reward-seeking behavior, and despair behavior.

    “we used a chronic unpredictable stress paradigm, which effectively induces anhedonia along with other depression-related behaviors [34–36], including reduced sensitivity to sucrose reward, decreased sex-related reward-seeking behavior, and despair behavior.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [34] Lu XY, Kim CS, Frazer A, Zhang W. Leptin: a potential novel antidepressant. Proc Natl Acad Sci USA. 2006;103:1593–8. source

      Not checked in the free run.

    • cites [35] Liu J, Perez SM, Zhang W, Lodge DJ, Lu XY. Selective deletion of the leptin receptor in dopamine neurons produces anxiogenic-like behavior and increases dopaminergic activity in amygdala. Mol Psychiatry. 2011;16:1024–38. source

      Not checked in the free run.

    • cites [36] Lei Y, Wang J, Wang D, Li C, Liu B, Fang X, et al. SIRT1 in forebrain excitatory neurons produces sexually dimorphic effects on depression-related behaviors and modulates neuronal excitability and synaptic transmission in the medial prefrontal cortex. Mol Psychiatry. 2020;25:1094–111. source

      Not checked in the free run.

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

    Premisecitation

    Irregular neuronal firing may be caused by intrinsic cellular mechanisms through the stochastic gating of ion channels in the membrane, such as potassium ion channels.

    “Second, irregular neuronal firing may be caused by intrinsic cellular mechanisms through the stochastic gating of ion channels in the membrane, such as potassium ion channels [70–73].”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [70] Kaffashian M, Shabani M, Goudarzi I, Behzadi G, Zali A, Janahmadi M. Profound alterations in the intrinsic excitability of cerebellar Purkinje neurons following neurotoxin 3-acetylpyridine (3-AP)-induced ataxia in rat: new insights into the role of small conductance K+ channels. Physiol Res. 2011;60:355–65. source

      Not checked in the free run.

    • cites [71] Mendonca PRF, Vargas-Caballero M, Erdelyi F, Szabo G, Paulsen O, Robinson HPC. Stochastic and deterministic dynamics of intrinsically irregular firing in cortical inhibitory interneurons. Elife. 2016;5:e16475. source

      Not checked in the free run.

    • cites [72] Dell’Orco JM, Pulst SM, Shakkottai VG. Potassium channel dysfunction underlies Purkinje neuron spiking abnormalities in spinocerebellar ataxia type 2. Hum Mol Genet. 2017;26:3935–45. source

      Not checked in the free run.

    • cites [73] Mendonca PRF, Kyle V, Yeo SH, Colledge WH, Robinson HPC. Kv4.2 channel activity controls intrinsic firing dynamics of arcuate kisspeptin neurons. J Physiol-Lond. 2018;596:885–99. source

      Not checked in the free run.

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

    Premisecitation

    The mesolimbic pathway has been implicated in depression.

    “However, available anatomical evidence suggests that AgRP neurons may functionally interact with the mesolimbic pathway [20, 62, 86], which has been implicated in depression [56].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [56] Nestler EJ, Carlezon WA. The mesolimbic dopamine reward circuit in depression. Biol Psychiatry. 2006;59:1151–9. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    The increase in amplitude but not frequency of spontaneous IPSCs suggests enhanced postsynaptic responsiveness.

    “The amplitude but not the frequency of spontaneous IPSCs was increased by chronic unpredictable stress, suggesting enhanced postsynaptic responsiveness.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    In the absence of synaptic transmission, AgRP neurons were capable of intrinsically firing.

    “In the absence of synaptic transmission, AgRP neurons were capable of intrinsically firing [65, 66].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [65] Paeger L, Karakasilioti I, Altmuller J, Frommole P, Bruning J, Kloppenburg P. Antagonistic modulation of NPY/AgRP and POMC neurons in the arcuate nucleus by noradrenalin. Elife. 2017;6:e25770. source

      Not checked in the free run.

    • cites [66] Cains S, Blomeley C, Kollo M, Racz R, Burdakov D. Agrp neuron activity is required for alcohol-induced overeating. Nat Commun. 2017;8:14014. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Chronic unpredictable stress may increase inhibitory synaptic transmission onto AgRP neurons partially through augmented postsynaptic sensitivity to synaptically released GABA, due either to increased function or increased number of postsynaptic GABA receptors clustered at synaptic sites.

    “These data suggest that chronic unpredictable stress may increase inhibitory synaptic transmission onto AgRP neurons partially through augmented postsynaptic sensitivity to synaptically released GABA, due either to increased function or increased number of postsynaptic GABA receptors clustered at synaptic sites [63, 64].”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [63] Otis TS, De Koninck Y, Mody I. Lasting potentiation of inhibition is associated with an increased number of gamma-aminobutyric acid type A receptors activated during miniature inhibitory postsynaptic currents. Proc Natl Acad Sci USA. 1994;91:7698–702. source

      Not checked in the free run.

    • cites [64] Nusser Z, Hajos N, Somogyi P, Mody I. Increased number of synaptic GABA(A) receptors underlies potentiation at hippocampal inhibitory synapses. Nature. 1998;395:172–7. source

      Not checked in the free run.

    • Rests on an assumption: Spontaneous IPSCs comprise both AP-dependent and independent inhibitory synaptic events.

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

    Premisecitation

    Available anatomical evidence suggests that AgRP neurons may functionally interact with the mesolimbic pathway.

    “However, available anatomical evidence suggests that AgRP neurons may functionally interact with the mesolimbic pathway [20, 62, 86], which has been implicated in depression [56].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [20] Bagnol D, Lu XY, Kaelin CB, Day HE, Ollmann M, Gantz I, et al. Anatomy of an endogenous antagonist: relationship between Agouti-related protein and proopiomelanocortin in brain. J Neurosci. 1999;19:RC26. source

      Not checked in the free run.

    • cites [62] Wang D, He X, Zhao Z, Feng Q, Lin R, Sun Y, et al. Whole-brain mapping of the direct inputs and axonal projections of POMC and AgRP neurons. Front Neuroanat. 2015;9:40. source

      Not checked in the free run.

    • cites [86] Broberger C, Johansen J, Johansson C, Schalling M, Hokfelt T. The neuropeptide Y/agouti gene-related protein (AGRP) brain circuitry in normal, anorectic, and monosodium glutamate-treated mice. Proc Natl Acad Sci USA. 1998;95:15043–8. source

      Not checked in the free run.

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

    Premisecitation

    The mesolimbic dopamine system, composed of dopamine neurons of the VTA and their projections to the NAc, is most often associated with the rewarding effects of food and sex.

    “The mesolimbic dopamine system, composed of dopamine neurons of the ventral tegmental area (VTA) and their projections to the nucleus accumbens (NAc), is most often associated with the rewarding effects of food and sex and plays an essential role in stress-induced anhedonia and other depression-related behavioral abnormalities [87, 88].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [87] Wise RA. Dopamine, learning and motivation. Nat Rev Neurosci. 2004;5:483–94. source

      Not checked in the free run.

    • cites [88] Russo SJ, Nestler EJ. The brain reward circuitry in mood disorders. Nat Rev Neurosci. 2013;14:609–25. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    This predominant excitation is reduced by chronic stress through both synaptic and intrinsic mechanisms.

    “This predominant excitation is reduced by chronic stress through both synaptic and intrinsic mechanisms.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    AHP currents are mainly mediated by calcium-dependent potassium channels, but also partly by calcium-independent potassium channels.

    “AHP currents are mainly mediated by calcium-dependent potassium channels [77, 78], but also partly by calcium-independent potassium channels [79].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [77] Lancaster B, Adams PR. Calcium-dependent current generating the afterhyperpolarization of hippocampal neurons. J Neurophysiol. 1986;55:1268–82. source

      Not checked in the free run.

    • cites [78] Hotson JR, Prince DA. A calcium-activated hyperpolarization follows repetitive firing in hippocampal neurons. J Neurophysiol. 1980;43:409–19. source

      Not checked in the free run.

    • cites [79] Lorenzon NM, Foehring RC. Relationship between repetitive firing and after hyperpolarizations in human neocortical neurons. J Neurophysiol. 1992;67:350–63. source

      Not checked in the free run.

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

    Premisecitation

    An increase in the irregularity of neuron firing reflects instability.

    “An increase in the irregularity of neuron firing reflects instability [67].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [67] Sakamoto K, Katori Y, Saito N, Yoshida S, Aihara K, Mushiake H. Increased firing irregularity as an emergent property of neural-state transition in monkey prefrontal cortex. PLoS ONE. 2013;8:e80906. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    AgRP neuronal activity may be necessary and sufficient for the manifestation of depression-related behaviors.

    “These results suggest that AgRP neuronal activity may be necessary and sufficient for the manifestation of depression-related behaviors.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    ATP-sensitive potassium channels were also detected in AgRP neurons.

    “ATP-sensitive potassium channels were also detected in AgRP neurons [61].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [61] van den Top M, Lee K, Whyment AD, Blanks AM, Spanswick D. Orexigen-sensitive NPY/AgRP pacemaker neurons in the hypothalamic arcuate nucleus. Nat Neurosci. 2004;7:493–4. source

      Not checked in the free run.

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

    Premisecitation

    Irregular neuronal firing can arise from fluctuating patterns of synaptic activity.

    “First, it can arise from fluctuating patterns of synaptic activity [68, 69].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [68] Destexhe A, Rudolph M, Fellous JM, Sejnowski TJ. Fluctuating synaptic conductances recreate in vivo-like activity in neocortical neurons. Neuroscience. 2001;107:13–24. source

      Not checked in the free run.

    • cites [69] Ribrault C, Sekimoto K, Triller A. From the stochasticity of molecular processes to the variability of synaptic transmission. Nat Rev Neurosci. 2011;12:375–87. source

      Not checked in the free run.

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

    Premisecitation

    AgRP neurons express high levels of SK channels.

    “AgRP neurons express high levels of SK channels [82].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [82] He Y, Shu G, Yang Y, Xu P, Xia Y, Wang C, et al. A small potassium current in AgRP/NPY neurons regulates feeding behavior and energy metabolism. Cell Rep. 2016;17:1807–18. source

      Not checked in the free run.

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

    Premisecitation

    One principal feedback mechanism in the control of the frequency and patterning of neuronal firing is afterhyperpolarization (AHP).

    “One principal feedback mechanism in the control of the frequency and patterning of neuronal firing is afterhyperpolarization, or AHP [74–76].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [74] Storm JF. Potassium currents in hippocampal pyramidal cells. Prog Brain Res. 1990;83:161–87. source

      Not checked in the free run.

    • cites [75] Saito Y, Takazawa T, Ozawa S. Relationship between afterhyperpolarization profiles and the regularity of spontaneous firings in rat medial vestibular nucleus neurons. Eur J Neurosci. 2008;28:288–98. source

      Not checked in the free run.

    • cites [76] Deister CA, Chan CS, Surmeier DJ, Wilson CJ. Calcium-activated SK channels influence voltage-gated ion channels to determine the precision of firing in globus pallidus neurons. J Neurosci. 2009;29:8452–61. source

      Not checked in the free run.

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

    Premisecitation

    Small-conductance calcium-activated potassium (SK) channels have been reported to control spontaneous firing regularity both in vivo and in vitro.

    “Particularly, small-conductance calcium-activated potassium (SK) channels have been reported to control the spontaneous firing regularity both in vivo and in vitro [80, 81].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [80] Pedarzani P, Stocker M. Molecular and cellular basis of small–and intermediate-conductance, calcium-activated potassium channel function in the brain. Cell Mol Life Sci. 2008;65:3196–217. source

      Not checked in the free run.

    • cites [81] Iyer R, Ungless MA, Faisal AA. Calcium-activated SK channels control firing regularity by modulating sodium channel availability in midbrain dopamine neurons. Sci Rep. 2017;7:5248. source

      Not checked in the free run.

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

    Premisecitation

    AgRP neurons in the ARC innervate the VTA.

    “AgRP neurons in the ARC have been shown to innervate the VTA and play a role in setting VTA dopamine neuronal activity and determining reward-associated behaviors [89].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [89] Dietrich MO, Bober J, Ferreira JG, Tellez LA, Mineur YS, Souza DO, et al. AgRP neurons regulate development of dopamine neuronal plasticity and nonfood-associated behaviors. Nat Neurosci. 2012;15:1108–10. source

      Not checked in the free run.

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

    Premisecitation

    Lesions of the BNST block the behavioral consequences of stress.

    “Lesions of this nucleus alter despair behavior [92] and block the behavioral consequences of stress [93].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [93] Hammack SE, Richey KJ, Watkins LR, Maier SF. Chemical lesion of the bed nucleus of the stria terminalis blocks the behavioral consequences of uncontrollable stress. Behav Neurosci. 2004;118:443–8. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    AgRP neurons in the ARC play a role in setting VTA dopamine neuronal activity and determining reward-associated behaviors.

    “AgRP neurons in the ARC have been shown to innervate the VTA and play a role in setting VTA dopamine neuronal activity and determining reward-associated behaviors [89].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [89] Dietrich MO, Bober J, Ferreira JG, Tellez LA, Mineur YS, Souza DO, et al. AgRP neurons regulate development of dopamine neuronal plasticity and nonfood-associated behaviors. Nat Neurosci. 2012;15:1108–10. source

      Not checked in the free run.

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

    Premisecitation

    Another candidate downstream target of AgRP neurons is the bed nucleus of the stria terminalis (BNST).

    “Another candidate downstream target of AgRP neurons is the bed nucleus of the stria terminalis (BNST) [20, 62], which has emerged as a key player in stress-related disorders [90, 91].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [20] Bagnol D, Lu XY, Kaelin CB, Day HE, Ollmann M, Gantz I, et al. Anatomy of an endogenous antagonist: relationship between Agouti-related protein and proopiomelanocortin in brain. J Neurosci. 1999;19:RC26. source

      Not checked in the free run.

    • cites [62] Wang D, He X, Zhao Z, Feng Q, Lin R, Sun Y, et al. Whole-brain mapping of the direct inputs and axonal projections of POMC and AgRP neurons. Front Neuroanat. 2015;9:40. source

      Not checked in the free run.

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

    Premisecitation

    The BNST has emerged as a key player in stress-related disorders.

    “Another candidate downstream target of AgRP neurons is the bed nucleus of the stria terminalis (BNST) [20, 62], which has emerged as a key player in stress-related disorders [90, 91].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [90] Hu P, Liu J, Maita I, Kwok C, Gu E, Gergues MM, et al. Chronic stress induces maladaptive behaviors by activating corticotropin-releasing hormone signaling in the mouse oval bed nucleus of the stria terminalis. J Neurosci. 2020;40:2519–37. source

      Not checked in the free run.

    • cites [91] Lebow MA, Chen A. Overshadowed by the amygdala: the bed nucleus of the stria terminalis emerges as key to psychiatric disorders. Mol Psychiatry. 2016;21:450–63. source

      Not checked in the free run.

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

    Premisecitation

    Lesions of the BNST alter despair behavior.

    “Lesions of this nucleus alter despair behavior [92] and block the behavioral consequences of stress [93].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [92] Schulz D, Canbeyli RS. Lesion of the bed nucleus of the stria terminalis enhances learned despair. Brain Res Bull. 2000;52:83–7. source

      Not checked in the free run.

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

    Premisecitation

    Phasic activation of VTA dopamine neurons reverses chronic mild/unpredictable stress-induced depression-like phenotypes, a phenomenon dependent on dopamine transmission in the NAc.

    “Selective inhibition of VTA dopamine neurons induces anhedonia and behavioral despair; conversely, phasic activation of VTA dopamine neurons reverses chronic mild/unpredictable stress-induced depression-like phenotypes, a phenomenon dependent on dopamine transmission in the NAc [41].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [41] Tye KM, Mirzabekov JJ, Warden MR, Ferenczi EA, Tsai HC, Finkelstein J, et al. Dopamine neurons modulate neural encoding and expression of depression-related behaviour. Nature. 2013;493:537–41. source

      Not checked in the free run.

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

    Premisecitation

    Selective inhibition of VTA dopamine neurons induces anhedonia and behavioral despair.

    “Selective inhibition of VTA dopamine neurons induces anhedonia and behavioral despair; conversely, phasic activation of VTA dopamine neurons reverses chronic mild/unpredictable stress-induced depression-like phenotypes, a phenomenon dependent on dopamine transmission in the NAc [41].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [41] Tye KM, Mirzabekov JJ, Warden MR, Ferenczi EA, Tsai HC, Finkelstein J, et al. Dopamine neurons modulate neural encoding and expression of depression-related behaviour. Nature. 2013;493:537–41. source

      Not checked in the free run.

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

    Premisecitation

    The mesolimbic dopamine system plays an essential role in stress-induced anhedonia and other depression-related behavioral abnormalities.

    “The mesolimbic dopamine system, composed of dopamine neurons of the ventral tegmental area (VTA) and their projections to the nucleus accumbens (NAc), is most often associated with the rewarding effects of food and sex and plays an essential role in stress-induced anhedonia and other depression-related behavioral abnormalities [87, 88].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [87] Wise RA. Dopamine, learning and motivation. Nat Rev Neurosci. 2004;5:483–94. source

      Not checked in the free run.

    • cites [88] Russo SJ, Nestler EJ. The brain reward circuitry in mood disorders. Nat Rev Neurosci. 2013;14:609–25. source

      Not checked in the free run.

71 of 71

  • The paper relies on this without support.

    The mechanisms underlying the development of hedonic deficits and other key symptoms of depression remain poorly understood.

    “Despite extensive research, the mechanisms underlying the development of hedonic deficits and other key symptoms of depression remain poorly understood.”

  • The paper relies on this without support.

    The functional roles of AgRP neurons in determining susceptibility to stress and the pathogenesis of depression remain unknown.

    “However, their functional roles in determining the susceptibility to stress and the pathogenesis of depression remain unknown.”

  • The paper relies on this without support.

    Spontaneous IPSCs comprise both AP-dependent and independent inhibitory synaptic events.

    “Spontaneous IPSCs comprise both AP-dependent and independent inhibitory synaptic events.”
3 of 3

Claims other papers make about this one

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

  • More severe, longer-lasting stress procedures produce anxiety- and depression-like phenotypes in these tasks.

    “This contrasts with more severe, longer-lasting stress procedures, which do produce anxiety- and depression-like phenotypes in these tasks80,100,101.”

    From A dual-pathway architecture for stress to disrupt agency and promote habit (2025) · cites this paper as [100]

Statistics that don’t add up

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

  • Inconsistent p-value: t(18) = 3.365, p = .004

    Minor

    The paper reports p = .004 here; recomputing from the test statistic and its degrees of freedom gives p = .003 (two-tailed). The check cannot tell whether the statistic or the p-value is the misprint.

    Reported p = .004 · recomputed p = .003

1 of 1

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(18) = 3.365reported p = .004 · recomputed p = .003

    “t(18) = 3.365, p = .004”

  • ConsistentF(1, 34) = 6.191reported p = .018 · recomputed p = .018

    “F(1,34) = 6.191, p = .018”

  • ConsistentF(1, 84) = 45.15reported p < .001 · recomputed p < .001

    “F(1,84) = 45.15, p < .001”

  • ConsistentF(1, 34) = 3.951reported p = .055 · recomputed p = .055

    “F(1,34) = 3.951, p = .055”

  • Consistentt(18) = 2.811reported p = .012 · recomputed p = .012

    “t(18) = 2.811, p = .012”

  • Consistentt(17) = 0.677reported p = .508 · recomputed p = .508

    “t(17) = 0.677, p = .508”

  • Consistentt(18) = 0.374reported p = .713 · recomputed p = .713

    “t(18) = 0.374, p = .713”

  • Consistentt(56) = 2.183reported p = .033 · recomputed p = .033

    “t(56) = 2.183, p = .033”

  • Consistentt(55) = 2.051reported p = .045 · recomputed p = .045

    “t(55) = 2.051, p = .045”

  • Consistentt(113) = 3.014reported p = .003 · recomputed p = .003

    “t(113) = 3.014, p = .003”

  • Consistentt(55) = 2.212reported p = .031 · recomputed p = .031

    “t(55) = 2.212, p = .031”

  • Consistentt(56) = 0.412reported p = .682 · recomputed p = .682

    “t(56) = 0.412, p = .682”

  • Consistentt(55) = 0.135reported p = .893 · recomputed p = .893

    “t(55) = 0.135, p = .893”

  • Consistentt(55) = 1.184reported p = .241 · recomputed p = .242

    “t(55) = 1.184, p = .241”

  • Consistentt(52) = 1.74reported p = .088 · recomputed p = .088

    “t(52) = 1.740, p = .088”

  • Consistentt(61) = 3.347reported p = .001 · recomputed p = .001

    “t(61) = 3.347, p = .001”

  • Consistentt(48) = 0.206reported p = .838 · recomputed p = .838

    “t(48) = 0.206, p = .838”

  • Consistentt(48) = 0.38reported p = .706 · recomputed p = .706

    “t(48) = 0.380, p = .706”

  • Consistentt(48) = 5.482reported p < .001 · recomputed p < .001

    “t(48) = 5.482, p < .001”

  • Consistentt(48) = 4.169reported p = .0001 · recomputed p < .001

    “t(48) = 4.169, p = .0001”

  • Consistentt(48) = 3.148reported p = .003 · recomputed p = .003

    “t(48) = 3.148, p = .003”

  • Consistentt(79) = 0.035reported p = .972 · recomputed p = .972

    “t(79) = 0.035, p = .972”

  • Consistentt(2) = 14.62reported p = .005 · recomputed p = .005

    “t(2) = 14.62, p = .005”

  • Consistentt(15) = 0.468reported p = .647 · recomputed p = .647

    “t(15) = 0.468, p = .647”

  • Consistentt(15) = 2.828reported p = .013 · recomputed p = .013

    “t(15) = 2.828, p = .013”

  • Consistentt(15) = 1.535reported p = .146 · recomputed p = .146

    “t(15) = 1.535, p = .146”

  • Consistentt(15) = 1.151reported p = .268 · recomputed p = .268

    “t(15) = 1.151, p = .268”

  • Consistentt(15) = 4.299reported p < .001 · recomputed p < .001

    “t(15) = 4.299, p < .001”

  • Consistentt(14) = 2.877reported p = .012 · recomputed p = .012

    “t(14) = 2.877, p = .012”

  • Consistentt(15) = 1.572reported p = .137 · recomputed p = .137

    “t(15) = 1.572, p = .137”

  • Consistentt(2) = 5.649reported p = .03 · recomputed p = .030

    “t(2) = 5.649, p = .030”

  • ConsistentF(2, 64) = 9.218reported p < .001 · recomputed p < .001

    “F(2,64) = 9.218, p < .001”

  • ConsistentF(1, 64) = 68.95reported p < .001 · recomputed p < .001

    “F(1,64) = 68.95, p < .001”

  • ConsistentF(2, 64) = 6.162reported p = .004 · recomputed p = .004

    “F(2,64) = 6.162, p = .004”

  • ConsistentF(2, 64) = 5.312reported p = .007 · recomputed p = .007

    “F(2,64) = 5.312, p = .007”

  • ConsistentF(1, 64) = 66.38reported p < .001 · recomputed p < .001

    “F(1,64) = 66.38, p < .001”

  • ConsistentF(2, 64) = 3.228reported p = .046 · recomputed p = .046

    “F(2,64) = 3.228, p = .046”

  • ConsistentF(2, 32) = 0.078reported p = .926 · recomputed p = .925

    “F(2,32) = 0.078, p = .926”

  • ConsistentF(2, 37) = 0.161reported p = .852 · recomputed p = .852

    “F(2,37) = 0.161, p = .852”

  • ConsistentF(2, 36) = 4.458reported p = .019 · recomputed p = .019

    “F(2,36) = 4.458, p = .019”

  • ConsistentF(2, 37) = 2.78reported p = .075 · recomputed p = .075

    “F(2,37) = 2.780, p = .075”

41 of 41

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.

  • Inconsistent degrees of freedom in the female urine sniffing ANOVA

    Minor

    The sniffing-object effect is reported with 84 denominator degrees of freedom, while the stress and interaction effects from the same two-way ANOVA have 34. The figure lists 19 mice, each tested with both sniffing objects, which is consistent with 34-not 84-residual degrees of freedom.

  • Female three-group ANOVA degrees of freedom exceed the listed sample

    Minor

    The listed female group sizes total 38 mice. An ordinary one-way ANOVA of those three groups would have 35 residual degrees of freedom, not the reported 37; the discrepancy occurs for both the pre-CUS sucrose-preference and locomotor analyses.

  • Silent-neuron percentage does not match its count

    Minor

    One of 24 control neurons is 4.17%, which rounds to 4% at the reported whole-percent precision, rather than 5%.

    Reported 5 · recomputed 4.166667

  • Female forced-swim ANOVA degrees of freedom exceed the listed sample

    Minor

    The listed female groups contain 38 mice altogether, giving 35 residual degrees of freedom for a three-group one-way ANOVA. The reported forced-swim result instead has 36.

4 of 4

Cites

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Authors
Fang, X., Jiang, S., Wang, J., Bai, Y., Kim, C. S., Blake, D., Weintraub, N. L., Lei, Y., & Lu, X.-Y.
Affiliations
  • Augusta University (US)
  • Binzhou Medical University (CN)
Corresponding author
Lu, Xin-Yun
ORCID
Venue
Molecular Psychiatry
Publisher
Springer Science and Business Media LLC
Published
2021-01-11
Volume
26
Issue
6
Pages
2299-2315
ISSN
  • 1359-4184 (ISSN-L)
  • 1476-5578
DOI
10.1038/s41380-020-01004-x
PMC
PMC8272726
OpenAlex
W3120352455
Semantic Scholar
a8e90dc1342ff816bbc412e6200c33ac0852edaf
Cited by
95
References
94
Citations by year
2026: 14 · 2025: 24 · 2024: 17 · 2023: 15 · 2022: 18 · 2021: 7
FWCI
7.50
Type
journal article
Language
en
Open access
Licence
Locations
  • Molecular Psychiatry (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, other-oa)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • U.S. Department of Health & Human Services | NIH | National Institute of Mental Health (MH100583, MH119456)
  • U.S. Department of Health & Human Services | NIH | National Institute on Aging (AG062166)
Topics
  • Life Sciences › Neuroscience › Endocrine and Autonomic Systems › Regulation of Appetite and Obesity
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neurotransmitter Receptor Influence on Behavior
  • Life Sciences › Neuroscience › Behavioral Neuroscience › Stress Responses and Cortisol
Keywords
chronic unpredictable mild stress, depression, AgRP neurons, arcuate nucleus, neuronal firing rate, synaptic inhibition, intrinsic excitability, chemogenetic manipulation, anhedonia, negative energy balance, synaptic plasticity, sex differences
MeSH terms
Animals, Arcuate Nucleus of Hypothalamus, Depression, Feeding Behavior, Female, Male, Neurons, Mice, Agouti-Related Protein

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.

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