Ethanol-induced conditioned place preference and aversion differentially alter plasticity in the bed nucleus of stria terminalis

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Contextual cues associated with drugs of abuse, such as ethanol, can trigger craving and drug-seeking behavior. Pavlovian procedures, such as place conditioning, have been widely used to study the rewarding/aversive properties of drugs and the association between environmental cues and drug seeking. Previous research has shown that ethanol as an unconditioned stimulus can induce a strong conditioned place preference (CPP) or aversion (CPA) in rodents. However, the neural mechanisms underlying ethanol-induced reward and aversion have not been thoroughly investigated. The bed nucleus of the stria terminalis (BNST), an integral part of the extended amygdala, is engaged by both rewarding and aversive stimuli and plays a role in ethanol-seeking behavior. Here, we used ex-vivo slice physiology to probe learning-induced synaptic plasticity in the BNST following ethanol-induced CPP and CPA. Male DBA/2 J mice (2–3 months old) were conditioned using previously reported ethanol-induced CPP/CPA procedures. Ethanol-induced CPP was associated with increased neuronal excitability in the ventral BNST (vBNST). Conversely, ethanol-induced CPA resulted in a significant decrease in spontaneous glutamatergic transmission without alterations in GABAergic signaling. Ethanol-CPA also led to a significant increase in the paired-pulse ratio at excitatory synapses, suggestive of a decrease in presynaptic glutamate release. Collectively, these data demonstrate that the vBNST is involved in the modulation of contextual learning associated with both the rewarding and the aversive properties of ethanol in mice.

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  1. 1.

    Anton RF. What is craving? Models and implications for treatment. Alcohol Res Health. 1999;23:165–73.

  2. 2.

    Krank MD, Wall A-M. Cue exposure during a period of abstinence reduces the resumption of operant behavior for oral ethanol reinforcement. Behav Neurosci. 1990;104:725–33.

  3. 3.

    Monti PM, Rohsenow DJ, Hutchison KE. Toward bridging the gap between biological, psychobiological and psychosocial models of alcohol craving. Addiction. 2002;95:229–36.

  4. 4.

    Crombag HS, Shaham Y. Renewal of drug seeking by contextual cues after prolonged extinction in rats. Behav Neurosci. 2002;116:169–73.

  5. 5.

    Zironi I, Burattini C, Aicardi G, Janak PH. Context is a trigger for relapse to alcohol. Behav Brain Res. 2006;167:150–5.

  6. 6.

    Morean ME, Corbin WR. Subjective response to alcohol: a critical review of the literature. Alcohol Clin Exp Res. 2010;34:385–95.

  7. 7.

    Schuckit MA. Low level of response to alcohol as a predictor of future alcoholism. Am J Psychiatry. 1994;151:184–9.

  8. 8.

    Schuckit MA, Smith TL, Anderson KG, Brown SA. Testing the level of response to alcohol: social information processing model of alcoholism risk—a 20-year prospective study. Alcohol Clin Exp Res. 2004;28:1881–9.

  9. 9.

    Verendeev A, Riley AL. Conditioned taste aversion and drugs of abuse: history and interpretation. Neurosci Biobehav Rev. 2012;36:2193–205.

  10. 10.

    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.

  11. 11.

    Vranjkovic O, Pina M, Kash TL, Winder DG. The bed nucleus of the stria terminalis in drug-associated behavior and affect: a circuit-based perspective. Neuropharmacology. 2017;122:100–6.

  12. 12.

    Dong H-W, Petrovich GD, Watts AG, Swanson LW. Basic organization of projections from the oval and fusiform nuclei of the bed nuclei of the stria terminalis in adult rat brain. J Comp Neurol. 2001a;436:430–55.

  13. 13.

    Dong H-W, Swanson LW. Projections from bed nuclei of the stria terminalis, anteromedial area: cerebral hemisphere integration of neuroendocrine, autonomic, and behavioral aspects of energy balance. J Comp Neurol. 2006a;494:142–78.

  14. 14.

    Dong H-W, Swanson LW. Projections from bed nuclei of the stria terminalis, dorsomedial nucleus: Implications for cerebral hemisphere integration of neuroendocrine, autonomic, and drinking responses. J Comp Neurol. 2006b;494:75–107.

  15. 15.

    Dong HW, Petrovich GD, Swanson LW. Topography of projections from amygdala to bed nuclei of the stria terminalis. Brain Res Rev. 2001b;38:192–46.

  16. 16.

    Poulin J-F, Arbour D, Laforest S, Drolet G. Neuroanatomical characterization of endogenous opioids in the bed nucleus of the stria terminalis. Prog Neuro-Psychopharmacol Biol Psychiatry. 2009;33:1356–65.

  17. 17.

    Sun N, Cassell MD. Intrinsic GABAergic neurons in the rat central extended amygdala. J Comp Neurol. 1993;330:381–404.

  18. 18.

    Canteras NS, Swanson LW. Projections of the ventral subiculum to the amygdala, septum, and hypothalamus: a PHAL anterograde tract‐tracing study in the rat. J Comp Neurol. 1992;324:180–94.

  19. 19.

    Cullinan WE, Herman JP, Watson SJ. Ventral subicular interaction with the hypothalamic paraventricular nucleus: Evidence for a relay in the bed nucleus of the stria terminalis. J Comp Neurol. 1993;332:1–20.

  20. 20.

    Vertes RP. Differential projections of the infralimbic and prelimbic cortex in the rat. Synapse. 2004;51:32–58.

  21. 21.

    Kudo T, Uchigashima M, Miyazaki T, Konno K, Yamasaki M, Yanagawa Y, et al. Three types of neurochemical projection from the bed nucleus of the stria terminalis to the ventral tegmental area in adult mice. J Neurosci. 2012;32:18035 LP–46.

  22. 22.

    Bardo MT, Bevins RA. Conditioned place preference: what does it add to our preclinical understanding of drug reward? Psychopharmacol (Berl). 2000;153:31–43.

  23. 23.

    Tzschentke TM. Measuring reward with the conditioned place preference paradigm: a comprehensive review of drug effects, recent progress and new issues. Prog Neurobiol. 1998;56:613–72.

  24. 24.

    Cunningham CL, Prather LK. Conditioning trial duration affects ethanol-induced conditioned place preference in mice. Anim Learn . 1992;20:187–94.

  25. 25.

    Ciccocioppo R, Panocka I, Froldi R, Quitadamo E, Massi M. Ethanol induces conditioned place preference in genetically selected alcohol-preferring rats. Psychopharmacology. 1999;141:235–41.

  26. 26.

    Morales M, Varlinskaya EI, Spear LP. Evidence for conditioned place preference to a moderate dose of ethanol in adult male Sprague–Dawley rats. Alcohol. 2012;46:643–8.

  27. 27.

    Risinger FO, Oakes RA. Dose- and conditioning trial-dependent ethanol-induced conditioned place preference in Swiss-Webster mice. Pharmacol Biochem Behav. 1996;55:117–23.

  28. 28.

    Cunningham CL, Okorn DM, Howard CE. Interstimulus interval determines whether ethanol produces conditioned place preference or aversion in mice. Anim Learn Behav. 1997;25:31–42.

  29. 29.

    Hill KG, Ryabinin AE, Cunningham CL. FOS expression induced by an ethanol-paired conditioned stimulus. Pharmacol Biochem Behav. 2007;87:208–21.

  30. 30.

    Mahler SV, Aston-Jones GS. Fos activation of selective afferents to ventral tegmental area during cue-induced reinstatement of cocaine seeking in rats. J Neurosci. 2012;32:13309–25.

  31. 31.

    Pina MM, Young EA, Ryabinin AE, Cunningham CL. The bed nucleus of the stria terminalis regulates ethanol-seeking behavior in mice. Neuropharmacology. 2015;99:627–38.

  32. 32.

    Sartor GC, Aston-Jones G. Regulation of the ventral tegmental area by the bed nucleus of the stria terminalis is required for expression of cocaine preference. Eur J Neurosci. 2012;36:3549–58.

  33. 33.

    Pina MM, Cunningham CL. Ethanol-seeking behavior is expressed directly through an extended amygdala to midbrain neural circuit. Neurobiol Learn Mem. 2017;137:83–91.

  34. 34.

    Kash TL, Baucum AJ II, Conrad KL, Colbran RJ, Winder DG. Alcohol exposure alters NMDAR function in the bed nucleus of the stria terminalis. Neuropsychopharmacology. 2009;34:2420.

  35. 35.

    Kash TL, Matthews RT, Winder DG. Alcohol inhibits NR2B-containing NMDA receptors in the ventral bed nucleus of the stria terminalis. Neuropsychopharmacology. 2007;33:1379.

  36. 36.

    Reisiger A-R, Kaufling J, Manzoni O, Cador M, Georges F, Caille S. Nicotine self-administration induces CB1-dependent LTP in the bed nucleus of the stria terminalis. J Neurosci. 2014;34:4285–92.

  37. 37.

    Dumont EC, Mark GP, Mader S, Williams JT. Self-administration enhances excitatory synaptic transmission in the bed nucleus of the stria terminalis. Nat Neurosci. 2005;8:413–4.

  38. 38.

    Cunningham CL, Gremel CM, Groblewski PA. Drug-induced conditioned place preference and aversion in mice. Nat Protoc. 2006;1:1662.

  39. 39.

    Mazzone CM, Pati D, Michaelides M, DiBerto J, Fox JH, Tipton G, et al. Acute engagement of Gq-mediated signaling in the bed nucleus of the stria terminalis induces anxiety-like behavior. Mol Psychiatry. 2016a;23:143.

  40. 40.

    Vezina P, Stewart J. Conditioned locomotion and place preference elicited by tactile cues paired exclusively with morphine in an open field. Psychopharmacol (Berl). 1987;91:375–80.

  41. 41.

    Cunningham CL. Localization of genes influencing ethanol-induced conditioned place preference and locomotor activity in BXD recombinant inbred mice. Psychopharmacol (Berl). 1995;120:28–41.

  42. 42.

    Kourrich S, Calu DJ, Bonci A. Intrinsic plasticity: an emerging player in addiction. Nat Rev Neurosci. 2015;16:173.

  43. 43.

    Zucker RS, Regehr WG. Short-term synaptic plasticity. Annu Rev Physiol. 2002;64:355–405.

  44. 44.

    Lüscher C, Malenka RC. Drug-evoked synaptic plasticity in addiction: from molecular changes to circuit remodeling. Neuron. 2011;69:650–63.

  45. 45.

    Cunningham CL, Henderson CM. Ethanol-induced conditioned place aversion in mice. Behav Pharmacol. 2000;11:591–602.

  46. 46.

    Cunningham CL, Tull LE, Rindal KE, Meyer PJ. Distal and proximal pre-exposure to ethanol in the place conditioning task: tolerance to aversive effect, sensitization to activating effect, but no change in rewarding effect. Psychopharmacol (Berl). 2002;160:414–24.

  47. 47.

    Dumont EC, Williams JT. Noradrenaline Triggers GABAA Inhibition of Bed Nucleus of the Stria Terminalis Neurons Projecting to the Ventral Tegmental Area. J Neurosci. 2004;24:8198 LP-8204.

  48. 48.

    Lüscher C, Slesinger PA. Emerging roles for G protein-gated inwardly rectifying potassium (GIRK) channels in health and disease. Nat Rev Neurosci. 2010;11:301.

  49. 49.

    Matsuda H, Saigusa A, Irisawa H. Ohmic conductance through the inwardly rectifying K channel and blocking by internal Mg2+. Nature. 1987;325:156.

  50. 50.

    Dascal N. Signalling via the G protein-activated K+channels. Cell Signal. 1997;9:551–73.

  51. 51.

    Aryal P, Dvir H, Choe S, Slesinger PA. A discrete alcohol pocket involved in GIRK channel activation. Nat Neurosci. 2009;12:988.

  52. 52.

    Herman MA, Sidhu H, Stouffer DG, Kreifeldt M, Le D, Cates-Gatto C, et al. GIRK3 gates activation of the mesolimbic dopaminergic pathway by ethanol. Proc Natl Acad Sci. 2015;112:7091 LP–6.

  53. 53.

    McCall NM, Kotecki L, Dominguez-Lopez S, Marron Fernandez de Velasco E, Carlblom N, Sharpe AL, et al. Selective ablation of girk channels in dopamine neurons alters behavioral effects of cocaine in mice. Neuropsychopharmacology. 2016;42:707.

  54. 54.

    Munoz MB, Padgett CL, Rifkin R, Terunuma M, Wickman K, Contet C, et al. A Role for the GIRK3 Subunit in methamphetamine-induced attenuation of gaba<sub>b</sub> receptor-activated GIRK currents in VTA dopamine neurons. J Neurosci. 2016;36:3106 LP–14.

  55. 55.

    Rifkin RA, Moss SJ, Slesinger PA. G protein-gated potassium channels: a link to drug addiction. Trends Pharmacol Sci. 2017;38:378–92.

  56. 56.

    Tipps ME, Raybuck JD, Kozell LB, Lattal KM, Buck KJ. G protein-gated inwardly rectifying potassium channel subunit 3 knock-out mice show enhanced ethanol reward. Alcohol Clin Exp Res. 2016;40:857–64.

  57. 57.

    Hill KG, Alva H, Blednov YA, Cunningham CL. Reduced ethanol-induced conditioned taste aversion and conditioned place preference in GIRK2 null mutant mice. Psychopharmacol (Berl). 2003;169:108–14.

  58. 58.

    Dong Y, Nasif FJ, Tsui JJ, Ju WY, Cooper DC, Hu X-T, et al. Cocaine-induced plasticity of intrinsic membrane properties in prefrontal cortex pyramidal neurons: adaptations in potassium currents. J Neurosci. 2005;25:936 LP–40.

  59. 59.

    Kamii H, Kurosawa R, Taoka N, Shinohara F, Minami M, Kaneda K. Intrinsic membrane plasticity via increased persistent sodium conductance of cholinergic neurons in the rat laterodorsal tegmental nucleus contributes to cocaine-induced addictive behavior. Eur J Neurosci. 2015;41:1126–38.

  60. 60.

    Kourrich S, Thomas MJ. Similar Neurons, opposite adaptations: psychostimulant experience differentially alters firing properties in accumbens core versus shell. J Neurosci. 2009;29:12275 LP–83.

  61. 61.

    Marcinkiewcz CA, Dorrier CE, Lopez AJ, Kash TL. Ethanol induced adaptations in 5-HT2c receptor signaling in the bed nucleus of the stria terminalis: Implications for anxiety during ethanol withdrawal. Neuropharmacology. 2015;89:157–67.

  62. 62.

    Pleil KE, Lowery-Gionta EG, Crowley NA, Li C, Marcinkiewcz CA, Rose JH, et al. Effects of chronic ethanol exposure on neuronal function in the prefrontal cortex and extended amygdala. Neuropharmacology. 2015a;99:735–49.

  63. 63.

    Francesconi W, Berton F, Repunte-Canonigo V, Hagihara K, Thurbon D, Lekic D, et al. Protracted withdrawal from alcohol and drugs of abuse impairs long-term potentiation of intrinsic excitability in the juxtacapsular bed nucleus of the stria terminalis. J Neurosci. 2009;29:5389 LP–401.

  64. 64.

    Kim S-Y, Adhikari A, Lee SY, Marshel JH, Kim CK, Mallory CS, et al. Diverging neural pathways assemble a behavioural state from separable features in anxiety. Nature. 2013;496:219.

  65. 65.

    Jennings JH, Sparta DR, Stamatakis AM, Ung RL, Pleil KE, Kash TL, et al. Distinct extended amygdala circuits for divergent motivational states. Nature. 2013;496:224.

  66. 66.

    Marcinkiewcz CA, Mazzone CM, D’Agostino G, Halladay LR, Hardaway JA, DiBerto JF, et al. Serotonin engages an anxiety and fear-promoting circuit in the extended amygdala. Nature. 2016;537:97.

  67. 67.

    Pleil KE, Rinker JA, Lowery-Gionta EG, Mazzone CM, McCall NM, Kendra AM, et al. NPY signaling inhibits extended amygdala CRF neurons to suppress binge alcohol drinking. Nat Neurosci. 2015b;18:545.

  68. 68.

    Silberman Y, Matthews RT, Winder DG. A corticotropin releasing factor pathway for ethanol regulation of the ventral tegmental area in the bed nucleus of the stria terminalis. J Neurosci. 2013;33:950 LP–60.

  69. 69.

    Lovinger DM, Kash TL. Mechanisms of neuroplasticity and ethanol’s effects on plasticity in the striatum and bed nucleus of the stria terminalis. Alcohol Res. 2015;37:109–24.

  70. 70.

    Mantsch JR, Baker DA, Funk D, Lê AD, Shaham Y. Stress-induced reinstatement of drug seeking: 20 years of progress. Neuropsychopharmacology. 2015;41:335.

  71. 71.

    Minami MBT-IR of N. Chapter 10 neuronal mechanisms for pain‐induced aversion: behavioral studies using a conditioned place Aversion. Test. 2009;85:135–44.

  72. 72.

    Malenka RC, Bear MF. LTP and LTD: an embarrassment of riches. Neuron. 2004;44:5–21.

  73. 73.

    McElligott ZA, Winder DG. Modulation of glutamatergic synaptic transmission in the bed nucleus of the stria terminalis. Prog Neuro-Psychopharmacol Biol Psychiatry. 2009;33:1329–35.

  74. 74.

    Harris NA, Winder DG. Synaptic plasticity in the bed nucleus of the stria terminalis: underlying mechanisms and potential ramifications for reinstatement of drug- and alcohol-seeking behaviors. ACS Chem Neurosci. 2018;9:2173–87.

  75. 75.

    Grueter BA, Gosnell HB, Olsen CM, Schramm-Sapyta NL, Nekrasova T, Landreth GE, et al. Extracellular-signal regulated kinase 1-dependent metabotropic glutamate receptor 5-induced long-term depression in the bed nucleus of the stria terminalis is disrupted by cocaine administration. J Neurosci. 2006;26:3210 LP–9.

  76. 76.

    Puente N, Cui Y, Lassalle O, Lafourcade M, Georges F, Venance L, et al. Polymodal activation of the endocannabinoid system in the extended amygdala. Nat Neurosci. 2011;14:1542.

  77. 77.

    Mazzone CM, Pati D, Michaelides M, DiBerto J, Fox JH, Tipton G, et al. (2016b). Acute engagement of Gq-mediated signaling in the bed nucleus of the stria terminalis induces anxiety-like behavior. Mol Psychiatry.

  78. 78.

    Delfs JM, Zhu Y, Druhan JP, Aston-Jones G. Noradrenaline in the ventral forebrain is critical for opiate withdrawal-induced aversion. Nature. 2000;403:430.

  79. 79.

    Deyama S, Katayama T, Ohno A, Nakagawa T, Kaneko S, Yamaguchi T, et al. Activation of the β-adrenoceptor–protein kinase a signaling pathway within the ventral bed nucleus of the stria terminalis mediates the negative affective component of pain in rats. J Neurosci. 2008;28:7728 LP–36.

  80. 80.

    Egli RE, Kash TL, Choo K, Savchenko V, Matthews RT, Blakely RD, et al. Norepinephrine modulates glutamatergic transmission in the bed nucleus of the stria terminalis. Neuropsychopharmacology. 2004;30:657.

  81. 81.

    McElligott ZA, Klug JR, Nobis WP, Patel S, Grueter BA, Kash TL, et al. Distinct forms of G<sub>q</sub>-receptor-dependent plasticity of excitatory transmission in the BNST are differentially affected by stress. Proc Natl Acad Sci. 2010;107:2271 LP–6.

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Funding and disclosure

This work was funded by NIAAA grants R01 AA019454 (TLK), U01 AA020911 (TLK), R01 AA025582 (TLK), P60 AA011605 (TLK) and F32 AA026485 (MMP). The authors declare no competing interests.

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Correspondence to Thomas L. Kash.

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