The serotonin (5-hydroxytryptamine (5-HT))-mediated system plays an important role in stress-related psychiatric disorders and substance abuse. Our previous studies showed that stress and drug exposure can modulate the dorsal raphe nucleus (DRN)-5-HT system via γ-aminobutyric acid (GABA)A receptors. Moreover, GABAA receptor-mediated inhibition of serotonergic DRN neurons is required for stress-induced reinstatement of opioid seeking.
Aim/methods:
To further test the role of GABAA receptors in the 5-HT system in stress and opioid-sensitive behaviors, our current study generated mice with conditional genetic deletions of the GABAA α1 subunit to manipulate GABAA receptors in either the DRN or the entire population of 5-HT neurons. The GABAA α1 subunit is a constituent of the most abundant GABAA subtype in the brain and the most highly expressed subunit in 5-HT DRN neurons.
Results:
Our results showed that mice with DRN-specific knockout of α1-GABAA receptors exhibited a normal phenotype in tests of anxiety- and depression-like behaviors as well as swim stress-induced reinstatement of morphine-conditioned place preference. By contrast, mice with 5-HT neuron-specific knockout of α1-GABAA receptors exhibited an anxiolytic phenotype at baseline and increased sensitivity to post-morphine withdrawal-induced anxiety.
Conclusions:
Our data suggest that GABAA receptors on 5-HT neurons contribute to anxiety-like behaviors and sensitivity of those behaviors to opioid withdrawal.
Alvarez-BagnarolYGarcíaRVendruscoloLF, et al. (2023) Inhibition of dorsal raphe GABAergic neurons blocks hyperalgesia during heroin withdrawal. Neuropsychopharmacology48: 1300–1308.
2.
AndradeTGZangrossiHGraeffFG (2013) The median raphe nucleus in anxiety revisited. J Psychopharmacol27: 1107–1115.
3.
ArainFMBoydKLGallagherMJ (2012) Decreased viability and absence-like epilepsy in mice lacking or deficient in the GABAA receptor α1 subunit. Epilepsia53: e161–e165.
4.
AtackJRHutsonPHCollinsonN, et al. (2005) Anxiogenic properties of an inverse agonist selective for alpha3 subunit-containing GABA A receptors. Br J Pharmacol144: 357–366.
5.
BarkerJSHinesRM (2020) Regulation of GABA receptor subunit expression in substance use disorders. Int J Mol Sci21: 4445.
6.
BrussaardABKitsKSBakerRE, et al. (1997) Plasticity in fast synaptic inhibition of adult oxytocin neurons caused by switch in GABA(A) receptor subunit expression. Neuron19: 1103–1114.
7.
CnopsVIyerVRParathyN, et al. (2022) Test, rinse, repeat: A review of carryover effects in rodent behavioral assays. Neurosci Biobehav Rev135: 104560.
CorteenNLCarterJARudolphU, et al. (2015) Localisation and stress-induced plasticity of GABAA receptor subunits within the cellular networks of the mouse dorsal raphe nucleus. Brain Struct Funct220: 2739–2763.
10.
CraigeCPLewandowskiSKirbyLG, et al. (2015) Dorsal raphe 5-HT(2C) receptor and GABA networks regulate anxiety produced by cocaine withdrawal. Neuropharmacology93: 41–51.
11.
DiasRSheppardWFFradleyRL, et al. (2005) Evidence for a significant role of alpha 3-containing GABAA receptors in mediating the anxiolytic effects of benzodiazepines. J Neurosci25: 10682–10688.
12.
EttenbergAOferOAMuellerCL, et al. (2011) Inactivation of the dorsal raphe nucleus reduces the anxiogenic response of rats running an alley for intravenous cocaine. Pharmacol Biochem Behav97: 632–639.
13.
FluyauDMitraPJainA, et al. (2022) Selective serotonin reuptake inhibitors in the treatment of depression, anxiety, and post-traumatic stress disorder in substance use disorders: a Bayesian meta-analysis. Eur J Clin Pharmacol78: 931–942.
14.
FuWLe MaîtreEFabreV, et al. (2010) Chemical neuroanatomy of the dorsal raphe nucleus and adjacent structures of the mouse brain. J Comp Neurol518: 3464–3494.
15.
GoeldnerCLutzPEDarcqE, et al. (2011) Impaired emotional-like behavior and serotonergic function during protracted abstinence from chronic morphine. Biol Psychiatry69: 236–244.
16.
HeikkiläATEchenkoOUusi-OukariM, et al. (2001) Morphine withdrawal increases expression of GABA(A) receptor epsilon subunit mRNA in locus coeruleus neurons. Neuroreport12: 2981–2985.
17.
Hernández-VázquezFGarduñoJHernández-LópezS (2019) GABAergic modulation of serotonergic neurons in the dorsal raphe nucleus. Rev Neurosci30: 289–303.
18.
HuangKWOchandarenaNEPhilsonAC, et al. (2019) Molecular and anatomical organization of the dorsal raphe nucleus. Elife8: e46464.
19.
HuangPLiu-ChenLYKirbyLG (2010) Anxiety-like effects of SR141716-precipitated delta9-tetrahydrocannabinol withdrawal in mice in the elevated plus-maze. Neurosci Lett475: 165–168.
20.
JacobsBLAzmitiaEC (1992) Structure and function of the brain serotonin system. Physiol Rev72: 165–229.
21.
JolasTNestlerEJAghajanianGK (2000) Chronic morphine increases GABA tone on serotonergic neurons of the dorsal raphe nucleus: association with an up-regulation of the cyclic AMP pathway. Neuroscience95: 433–443.
22.
KirbyLGFreeman-DanielsELemosJC, et al. (2008) Corticotropin-releasing factor increases GABA synaptic activity and induces inward current in 5-hydroxytryptamine dorsal raphe neurons. J Neurosci28: 12927–12937.
23.
KirbyLGZeebFDWinstanleyCA (2011) Contributions of serotonin in addiction vulnerability. Neuropharmacology61: 421–432.
24.
KoobGF (2000) Neurobiology of addiction. Toward the development of new therapies. Ann N Y Acad Sci909: 170–185.
25.
KoobGF (2021) Drug addiction: Hyperkatifeia/negative reinforcement as a framework for medications development. Pharmacol Rev73: 163–201.
26.
KoobGFVolkowND (2016) Neurobiology of addiction: A neurocircuitry analysis. Lancet Psychiatry3: 760–773.
27.
KralicJEKorpiERO’BuckleyTK, et al. (2002). Molecular and pharmacological characterization of GABA(A) receptor alpha1 subunit knockout mice. J Pharmacol Exp Ther302: 1037–1045.
28.
KralicJESidlerCParpanF, et al. (2006) Compensatory alteration of inhibitory synaptic circuits in cerebellum and thalamus of gamma-aminobutyric acid type A receptor alpha1 subunit knockout mice. J Comp Neurol495: 408–421.
29.
LemosJCZhangGWalshT, et al. (2011) Stress-hyperresponsive WKY rats demonstrate depressed dorsal raphe neuronal excitability and dysregulated CRF-mediated responses. Neuropsychopharmacology36: 721–734.
30.
LiCKirbyLG (2016) Effects of cocaine history on postsynaptic GABA receptors on dorsal raphe serotonin neurons in a stress-induced relapse model in rats. Eur Neuropsychopharmacol26: 45–54.
31.
LiCMcCloskeyNPhillipsJ, et al. (2021) CRF-5-HT interactions in the dorsal raphe nucleus and motivation for stress-induced opioid reinstatement. Psychopharmacology (Berl)238: 29–40.
32.
LiCStaubDRKirbyLG (2013) Role of GABA receptors in dorsal raphe nucleus in stress-induced reinstatement of morphine-conditioned place preference in rats. Psychopharmacology (Berl)230: 537–545.
33.
ListerRG (1987) The use of a plus-maze to measure anxiety in the mouse. Psychopharmacology (Berl)92: 180–185.
34.
LutzPEPradhanAAGoeldnerC, et al. (2011) Sequential and opposing alterations of 5-HT(1A) receptor function during withdrawal from chronic morphine. Eur Neuropsychopharmacol21: 835–840.
35.
MaHWangNWangX, et al. (2018) Wnt7a in mouse insular cortex contributes to anxiety-like behavior during protracted abstinence from morphine. Neuroscience394: 164–176.
36.
MantschJRBakerDAFunkD, et al. (2016) Stress-induced reinstatement of drug seeking: 20 years of progress. Neuropsychopharmacology41: 335–356.
37.
McKernanRMWhitingPJ (1996) Which GABAA-receptor subtypes really occur in the brain?Trends Neurosci19: 139–143.
38.
MonroeSCRadkeAK (2023) Opioid withdrawal: Role in addiction and neural mechanisms. Psychopharmacology (Berl)240: 1417–1433.
39.
MüllerCPHombergJR (2015) The role of serotonin in drug use and addiction. Behav Brain Res277: 146–192.
40.
NietzAKrook-MagnusonCGutierrezH, et al. (2020) Selective loss of the GABA. J Neurophysiol124: 1183–1197.
41.
NygardSKHourguettesNJSobczakGG, et al. (2016) Stress-induced reinstatement of nicotine preference requires dynorphin/kappa opioid activity in the basolateral amygdala. J Neurosci36: 9937–9948.
42.
OkatyBWCommonsKGDymeckiSM (2019) Embracing diversity in the 5-HT neuronal system. Nat Rev Neurosci20: 397–424.
43.
PaxinosGFranklinKBJ (2001) The Mouse Brain in Stereotaxic Coordinates (2nd edn). San Diego: Academic Press.
44.
PomrenzeMBCardozo PintoDFNeumannPA, et al. (2022) Modulation of 5-HT release by dynorphin mediates social deficits during opioid withdrawal. Neuron110: 4125.e6–4143.e6.
45.
PorsoltRDLepichonMJalfreM (1977) Depression - new animal-model sensitive to antidepressant treatments. Nature266: 730–732.
46.
PortugalGSAl-HasaniRFakiraAK, et al. (2014) Hippocampal long-term potentiation is disrupted during expression and extinction but is restored after reinstatement of morphine place preference. J Neurosci34: 527–538.
47.
PourhamzehMMoravejFGArabiM, et al. (2022) The roles of serotonin in neuropsychiatric disorders. Cell Mol Neurobiol42: 1671–1692.
48.
ReynoldsDSO’MearaGFNewmanRJ, et al. (2003) GABA(A) alpha 1 subunit knock-out mice do not show a hyperlocomotor response following amphetamine or cocaine treatment. Neuropharmacology44: 190–198.
49.
ShiXvon WeltinEFitzsimmonsE, et al. (2022) Reactivation of cocaine contextual memory engages mechanistic target of rapamycin/S6 kinase 1 signaling. Front Pharmacol13: 976932.
50.
SpijkerSHoutzagerSWDe GunstMC, et al. (2004) Morphine exposure and abstinence define specific stages of gene expression in the rat nucleus accumbens. FASEB J18: 848–850.
51.
StaubDRLundenJWCathelAM, et al. (2012) Morphine history sensitizes postsynaptic GABA receptors on dorsal raphe serotonin neurons in a stress-induced relapse model in rats. Psychoneuroendocrinology37: 859–870.
52.
SurCWaffordKAReynoldsDS, et al. (2001) Loss of the major GABA(A) receptor subtype in the brain is not lethal in mice. J Neurosci21: 3409–3418.
53.
TakahashiAKwaCDeboldJF, et al. (2010) GABA(A) receptors in the dorsal raphé nucleus of mice: escalation of aggression after alcohol consumption. Psychopharmacology (Berl)211: 467–477.
54.
TaoRAuerbachSB (2005) mu-Opioids disinhibit and kappa-opioids inhibit serotonin efflux in the dorsal raphe nucleus. Brain Res1049: 70–79.
55.
TarachaE (2021) The role of serotoninergic system in psychostimulant effects. Postep Psychiatr Neurol30: 258–269.
56.
ValentinoRJLuckiIVanBE (2010) Corticotropin-releasing factor in the dorsal raphe nucleus: Linking stress coping and addiction. Brain Res1314: 29–37.
57.
WelschLBaillyJDarcqE, et al. (2020) The negative affect of protracted opioid abstinence: Progress and perspectives from rodent models. Biol Psychiatry87: 54–63.
58.
XuWWangYMaZ, et al. (2013) l-Isocorypalmine reduces behavioral sensitization and rewarding effects of cocaine in mice by acting on dopamine receptors. Drug Alcohol Depend133: 693–703.
59.
YalcinIBelzungCSurgetA (2008) Mouse strain differences in the unpredictable chronic mild stress: A four-antidepressant survey. Behav Brain Res193: 140–143.
60.
ZangrossiHGraeffFG (2014) Serotonin in anxiety and panic: contributions of the elevated T-maze. Neurosci Biobehav Rev46: 397–406.
61.
ZanosPGeorgiouPGonzalezLR, et al. (2016) Emotional impairment and persistent upregulation of mGlu5 receptor following morphine abstinence: Implications of an mGlu5-MOPr interaction. Int J Neuropsychopharmacol19: pyw011.
62.
ZanosPGeorgiouPWrightSR, et al. (2014) The oxytocin analogue carbetocin prevents emotional impairment and stress-induced reinstatement of opioid-seeking in morphine-abstinent mice. Neuropsychopharmacology39: 855–865.
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