**Author details**

Melanie M. Pina\* and Amy R. Williams

\*Address all correspondence to: melareaoftsai@gmail.com

Oregon Health & Science University, 3181 SW Sam Jackson Park Rd, Portland, United States

### **References**


[4] Ciccocioppo R, Angeletti S, Weiss F. Long-lasting resistance to extinction of response reinstatement induced by ethanol-related stimuli: role of genetic ethanol preference. Alcohol Clin Exp Res. 2001 Oct;25(10):1414–9.

despite receiving treatment for alcohol abuse. This chapter has outlined the animal models that are being used by preclinical researchers to better understand the formation, expression, extinction, and reinstatement (relapse) of alcohol cue associations that promote ethanol seeking and has summarized each of their advantages and disadvantages. Of particular use is the CPP paradigm. CPP allows the experimenter to separate distinct phases of acquiring, expressing, extinguishing, and reexpressing conditioned ethanol seeking and thus can easily study the neural mechanisms involved in each. This chapter also presented the classical and contemporary tools that can be used separately and in conjunction to probe the exact neural structures and circuitry involved in alcohol cues and seeking. Although classical tools have given us the greatest insight into the neurobiology of ethanol seeking thus far, contemporary tools have been and will allow for a much clearer and specific understanding of the structures involved in animal models of alcohol seeking. Finally, this chapter presented evidence from ethanol self-administration and ethanol CPP studies of the modulation of ethanol seeking by the mesolimbic structures (VTA, NAc), the limbic system (amygdala, BNST, ACC), and cortical structures (mPFC). Of particular importance is the VTA that sends vast dopaminergic input to many of these structures. The challenge of future research is to identify more structures critical for the acquisition and expression, and especially the extinction and reinstatement, of ethanol CPP and the inputs of the VTA that modulate dopaminergic tone and thus ethanolseeking behavior (such as the BNST-VTA projection). A better understanding of the whole circuit driving every aspect of ethanol seeking will improve our knowledge of AUDs and

Oregon Health & Science University, 3181 SW Sam Jackson Park Rd, Portland, United States

[1] Moos RH, Moos BS. Rates and predictors of relapse after natural and treated remission

[2] Ciccocioppo R, Martin-Fardon R, Weiss F. Stimuli associated with a single cocaine experience elicit long-lasting cocaine-seeking. Nat Neurosci. 2004 Mar 28;7(5):495–6.

[3] Ciccocioppo R, Sanna PP, Weiss F. Cocaine-predictive stimulus induces drug-seeking behavior and neural activation in limbic brain regions after multiple months of abstinence: reversal by D(1) antagonists. Proc Natl Acad Sci U S A 2001 Feb;98(4):1976–

from alcohol use disorders. Addiction 2006 Feb;101(2):212–22.

treatment options.

**Author details**

Melanie M. Pina\*

**References**

81.

and Amy R. Williams

68 Recent Advances in Drug Addiction Research and Clinical Applications

\*Address all correspondence to: melareaoftsai@gmail.com


[32] van der Kooy D, O'Shaughnessy M, Mucha RF, Kalant H. Motivational properties of ethanol in naive rats as studied by place conditioning. Pharmacol Biochem Behav. 1983 Sep;19(3):441–5.

[18] Epstein DH, Preston KL, Stewart J, Shaham Y. Toward a model of drug relapse: an assessment of the validity of the reinstatement procedure. Psychopharmacology (Berl).

[19] Childs E, de Wit H. Amphetamine-induced place preference in humans. Biol Psychiatry

[20] Tzschentke TM. Measuring reward with the conditioned place preference paradigm: a comprehensive review of drug effects, recent progress and new issues. Prog Neurobiol.

[21] Tzschentke TM. Measuring reward with the conditioned place preference (CPP) paradigm: update of the last decade. Addict Biol. 2007 Sep;12(3–4):227–462.

[22] Bormann NM, Cunningham CL. Ethanol-induced conditioned place aversion in rats: effect of interstimulus interval. Pharmacol Biochem Behav. 1998 Feb;59(2):427–32. [23] Cunningham CL, Henderson CM. Ethanol-induced conditioned place aversion in mice.

[24] Cunningham CL, Clemans JM, Fidler TL. Injection timing determines whether intra‐ gastric ethanol produces conditioned place preference or aversion in mice. Pharmacol

[25] Cunningham CL, Ferree NK, Howard MA. Apparatus bias and place conditioning with

[26] Cunningham CL, Okorn DM, Howard CE. Interstimulus interval determines whether ethanol produces conditioned place preference or aversion in mice. Anim Learn Behav.

[27] Grahame NJ, Chester JA, Rodd-Henricks K. Alcohol place preference conditioning in high-and low-alcohol preferring selected lines of mice. Pharmacol Biochem Behav. 2001

[28] Ciccocioppo R, Panocka I, Froldi R, Quitadamo E, Massi M. Ethanol induces condi‐ tioned place preference in genetically selected alcohol-preferring rats. Psychopharma‐

[29] 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 Nov;46(7):

[30] Asin KE, Wirtshafter D, Tabakoff B. Failure to establish a conditioned place preference

[31] Cunningham CL. Flavor and location aversions produced by ethanol. Behav Neural

with ethanol in rats. Pharmacol Biochem Behav. 1985 Feb;22(2):169–73.

ethanol in mice. Psychopharmacology (Berl). 2003 Dec;170(4):409–22.

2006 Sep 22;189(1):1–16.

70 Recent Advances in Drug Addiction Research and Clinical Applications

2009 May 15;65(10):900–4.

1998 Dec;56(6):613–72.

1997;25:31–42.

Apr;68(4):805–14.

643–8.

Behav Pharmacol. 2000 Nov;11(7–8):591–602.

Biochem Behav. 2002 Jun;72(3):659–68.

cology (Berl). 1999 Jan;141(3):235–41.

Biol. 1979 Nov;27(3):362–7.


[59] Di Ciano P, Everitt BJ. Direct interactions between the basolateral amygdala and nucleus accumbens core underlie cocaine-seeking behavior by rats. J Neurosci. 2004 Aug 11;24(32):7167–73.

[45] McCall JG, Kim T-I, Shin G, Huang X, Jung YH, Al-Hasani R, et al. Fabrication and application of flexible, multimodal light-emitting devices for wireless optogenetics. Nat

[46] Park SI, Shin G, Banks A, McCall JG, Siuda ER, Schmidt MJ, et al. Ultraminiaturized photovoltaic and radio frequency powered optoelectronic systems for wireless

[47] Gysbrechts B, Wang L, Trong NND, Cabral H, Navratilova Z, Battaglia F, et al. Light distribution and thermal effects in the rat brain under optogenetic stimulation. J

[48] Reig R, Mattia M, Compte A, Belmonte C, Sanchez-Vives MV. Temperature modulation of slow and fast cortical rhythms. J Neurophysiol. 2010 Mar;103(3):1253–61.

[49] Stujenske JM, Spellman T, Gordon JA. Modeling the spatiotemporal dynamics of light and heat propagation for in vivo optogenetics. Cell Rep. 2015 Jul 21;12(3):525–34. [50] Sternson SM, Roth BL. Chemogenetic tools to interrogate brain functions. Annu Rev

[51] Armbruster BN, Li X, Pausch MH, Herlitze S, Roth BL. Evolving the lock to fit the key to create a family of G protein-coupled receptors potently activated by an inert ligand.

[52] Alexander GM, Rogan SC, Abbas AI, Armbruster BN, Pei Y, Allen JA, et al. Remote control of neuronal activity in transgenic mice expressing evolved G protein-coupled

[53] Nichols CD, Roth BL. Engineered G-protein coupled receptors are powerful tools to investigate biological processes and behaviors. Front Mol Neurosci. 2009;2:16.

[54] Bender D, Holschbach M, Stöcklin G. Synthesis of n.c.a. carbon-11 labelled clozapine and its major metabolite clozapine-N-oxide and comparison of their biodistribution in

[55] Farrell MS, Roth BL. Pharmacosynthetics: reimagining the pharmacogenetic approach.

[56] Guettier J-M, Gautam D, Scarselli M, Ruiz de Azua I, Li JH, Rosemond E, et al. A chemical-genetic approach to study G protein regulation of beta cell function in vivo.

[57] Vardy E, Robinson JE, Li C, Olsen RHJ, DiBerto JF, Giguere PM, et al. A New DREADD facilitates the multiplexed chemogenetic interrogation of behavior. Neuron 2015 May

[58] Bechtholt AJ, Gremel CM, Cunningham CL. Handling blocks expression of conditioned place aversion but not conditioned place preference produced by ethanol in mice.

Protoc. 2013 Dec;8(12):2413–28.

72 Recent Advances in Drug Addiction Research and Clinical Applications

optogenetics. J Neural Eng. 2015 Oct;12(5):056002.

Proc Natl Acad Sci U S A 2007 Mar 20;104(12):5163–8.

receptors. Neuron 2009 Jul 16;63(1):27–39.

mice. Nucl Med Biol. 1994 Oct;21(7):921–5.

Proc Natl Acad Sci U S A 2009 Nov 10;106(45):19197–202.

Pharmacol Biochem Behav. 2004 Dec;79(4):739–44.

Brain Res. 2013 May;1511:6–20.

20;86(4):936–46.

Biophoton. 2015 Jul;(Epub ahead of print)

Neurosci. 2014 Jun;37(1):387–407.


[86] Fuchs RA, Ramirez DR, Bell GH. Nucleus accumbens shell and core involvement in drug context-induced reinstatement of cocaine seeking in rats. Psychopharmacology (Berl). 2008 Nov;200(4):545–56.

[72] McDaid J, McElvain MA, Brodie MS. Ethanol effects on dopaminergic ventral tegmen‐ tal area neurons during block of Ih: involvement of barium-sensitive potassium

[73] Morikawa H, Morrisett RA. Ethanol action on dopaminergic neurons in the ventral tegmental area: interaction with intrinsic ion channels and neurotransmitter inputs. Int

[74] Mrejeru A, Martí-Prats L, Avegno EM, Harrison NL, Sulzer D. A subset of ventral tegmental area dopamine neurons responds to acute ethanol. Neuroscience 2015 Apr

[75] Dayas CV, Liu X, Simms JA, Weiss F. Distinct patterns of neural activation associated with ethanol seeking: effects of naltrexone. Biol Psychiatry 2007 Apr;61(8):979–89.

[76] Swanson LW. The projections of the ventral tegmental area and adjacent regions: a combined fluorescent retrograde tracer and immunofluorescence study in the rat. Brain

[77] Cador M, Robbins TW, Everitt BJ. Involvement of the amygdala in stimulus-reward associations: interaction with the ventral striatum. Neuroscience 1989;30(1):77–86.

[78] Clark JJ, Collins AL, Sanford CA, Phillips PEM. Dopamine encoding of Pavlovian incentive stimuli diminishes with extended training. J Neurosci. 2013 Feb 20;33(8):3526–

[79] Di Chiara G, Bassareo V, Fenu S, De Luca MA, Spina L, Cadoni C, et al. Dopamine and drug addiction: the nucleus accumbens shell connection. Neuropharmacology

[80] Ikemoto S, Panksepp J. The role of nucleus accumbens dopamine in motivated behavior: a unifying interpretation with special reference to reward-seeking. Brain Res

[81] Wise RA. Dopamine, learning and motivation. Nat Rev Neurosci. 2004 Jun;5(6):483–94.

[82] Everitt BJ, Robbins TW. Neural systems of reinforcement for drug addiction: from

[83] McFarland K, Kalivas PW. The circuitry mediating cocaine-induced reinstatement of

[84] Alderson HL, Parkinson JA, Robbins TW, Everitt BJ. The effects of excitotoxic lesions of the nucleus accumbens core or shell regions on intravenous heroin self-administra‐

[85] Fuchs RA, Evans KA, Parker MC, See RE. Differential involvement of the core and shell subregions of the nucleus accumbens in conditioned cue-induced reinstatement of cocaine seeking in rats. Psychopharmacology (Berl). 2004 Nov;176(3–4):459–65.

actions to habits to compulsion. Nat Neurosci. 2005 Nov;8(11):1481–9.

drug-seeking behavior. J Neurosci. 2001 Nov 1;21(21):8655–63.

tion in rats. Psychopharmacology (Berl). 2000 Dec 21;153(4):455–63.

currents. J Neurophysiol. 2008 Jun 25;100(3):1202–10.

Rev Neurobiol. 2010;91:235–88.

74 Recent Advances in Drug Addiction Research and Clinical Applications

Res Bull. 1982 Jul;9(1–6):321–53.

2004;47(Suppl. 1):227–41.

Brain Res Rev. 1999 Dec;31(1):6–41.

2;290:649–58.

32.


[110] Young EA, Dreumont SE, Cunningham CL. Role of nucleus accumbens dopamine receptor subtypes in the learning and expression of alcohol-seeking behavior. Neuro‐ biol Learn Mem. 2014 Jun;108:28–37.

[98] Simms JA, Haass-Koffler CL, Bito-Onon J, Li R, Bartlett SE. Mifepristone in the central nucleus of the amygdala reduces yohimbine stress-induced reinstatement of ethanol-

[99] Zhao Y, Dayas CV, Aujla H, Baptista MAS, Martin-Fardon R, Weiss F. Activation of group II metabotropic glutamate receptors attenuates both stress and cue-induced ethanol-seeking and modulates c-fos expression in the hippocampus and amygdala. J

[100] Corbit LH, Nie H, Janak PH. Habitual alcohol seeking: time course and the contribution of subregions of the dorsal striatum. Biol Psychiatry 2012 Sep;72(5):389–95.

[101] Corbit LH, Nie H, Janak PH. Habitual responding for alcohol depends upon both AMPA and D2 receptor signaling in the dorsolateral striatum. Front Behav Neurosci.

[102] Brown RM, Kim AK, Khoo SY-S, Kim JH, Jupp B, Lawrence AJ. Orexin-1 receptor signalling in the prelimbic cortex and ventral tegmental area regulates cue-induced reinstatement of ethanol-seeking in iP rats. Addict Biol. 2015 May;21(3):603–12. [103] Hauser SR, Deehan GA, Toalston JE, Bell RL, McBride WJ, Rodd ZA. Enhanced alcoholseeking behavior by nicotine in the posterior ventral tegmental area of female alcoholpreferring (P) rats: modulation by serotonin-3 and nicotinic cholinergic receptors.

[104] Hauser SR, Ding Z-M, Getachew B, Toalston JE, Oster SM, McBride WJ, et al. The posterior ventral tegmental area mediates alcohol-seeking behavior in alcohol-

[105] Löf E, Olausson P, deBejczy A, Stomberg R, McIntosh JM, Taylor JR, et al. Nicotinic acetylcholine receptors in the ventral tegmental area mediate the dopamine activating and reinforcing properties of ethanol cues. Psychopharmacology (Berl). 2007 Aug

[106] Fidler TL, Bakner L, Cunningham CL. Conditioned place aversion induced by intra‐ gastric administration of ethanol in rats. Pharmacol Biochem Behav. 2004 Apr;77(4):

[107] Cunningham CL. Localization of genes influencing ethanol-induced conditioned place preference and locomotor activity in BXD recombinant inbred mice. Psychopharma‐

[108] Cunningham CL, Niehus DR, Malott DH, Prather LK. Genetic differences in the rewarding and activating effects of morphine and ethanol. Psychopharmacology (Berl).

[109] Gremel CM, Cunningham CL. Roles of the nucleus accumbens and amygdala in the acquisition and expression of ethanol-conditioned behavior in mice. J Neurosci.

seeking. Neuropsychopharmacology 2012 Mar;37(4):906–18.

Psychopharmacology (Berl). 2014 Mar 6;231(18):3745–55.

preferring rats. J Pharmacol Exp Ther. 2011 Mar;336(3):857–65.

Neurosci. 2006 Sep 27;26(39):9967–74.

76 Recent Advances in Drug Addiction Research and Clinical Applications

2014;8:301.

17;195(3):333–43.

cology (Berl). 1995 Jul;120(1):28–41.

1992 Jun;107(2–3):385–93.

2008;28(5):1076–84.

731–43.


[137] Kudo T, Konno K, Uchigashima M, Yanagawa Y, Sora I, Minami M, et al. GABAergic neurons in the ventral tegmental area receive dual GABA/enkephalin-mediated inhibitory inputs from the bed nucleus of the stria terminalis. Eur J Neurosci. Jun;39(11): 1796–809.

[123] Groblewski PA, Ryabinin AE, Cunningham CL. Activation and role of the medial prefrontal cortex (mPFC) in extinction of ethanol-induced associative learning in mice.

[124] Geisler S, Derst C, Veh RW, Zahm DS. Glutamatergic afferents of the ventral tegmental

[125] Omelchenko N, Sesack SR. Glutamate synaptic inputs to ventral tegmental area neurons in the rat derive primarily from subcortical sources. Neuroscience 2007;146(3):

[126] Carr DB, Sesack SR. Projections from the rat prefrontal cortex to the ventral tegmental area: target specificity in the synaptic associations with mesoaccumbens and meso‐

[127] Overton PG, Clark D. Burst firing in midbrain dopaminergic neurons. Brain Res Brain

[128] Wanat MJ, Willuhn I, Clark JJ, Phillips PEM. Phasic dopamine release in appetitive behaviors and drug addiction. Curr Drug Abuse Rev. 2009;2(2):195–213.

[129] Jennings JH, Sparta DR, Stamatakis AM, Ung RL, Pleil KE, Kash TL, et al. Distinct extended amygdala circuits for divergent motivational states. Nature 2013 Apr

[130] Lammel S, Lim BK, Ran C, Huang KW, Betley MJ, Tye KM, et al. Input-specific control of reward and aversion in the ventral tegmental area. Nature 2012 Nov;491(7423):212–

[131] Omelchenko N, Sesack SR. Laterodorsal tegmental projections to identified cell populations in the rat ventral tegmental area. J Comp Neurol. 2005;483(2):217–35.

[132] Lodge DJ, Grace AA. The laterodorsal tegmentum is essential for burst firing of ventral tegmental area dopamine neurons. Proc Natl Acad Sci U S A 2006;103(13):5167–72.

[133] Georges F, Aston-Jones GS. Potent regulation of midbrain dopamine neurons by the bed nucleus of the stria terminalis. J Neurosci. 2001 Aug 15;21(16):RC160.

[134] Georges F, Aston-Jones GS. Activation of ventral tegmental area cells by the bed nucleus of the stria terminalis: a novel excitatory amino acid input to midbrain

[135] Jalabert M, Aston-Jones GS, Herzog E, Manzoni O, Georges F. Role of the bed nucleus of the stria terminalis in the control of ventral tegmental area dopamine neurons. Prog

[136] 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 Dec;32(50):18035–46.

Neuro-Psychopharmacol Biol Psychiatry 2009 Nov 13;33(8):1336–46.

dopamine neurons. J Neurosci. 2002 Jun;22(12):5173–87.

Neurobiol Learn Mem. 2012 Jan;97(1):37–46.

78 Recent Advances in Drug Addiction Research and Clinical Applications

1259–74.

area in the rat. J Neurosci. 2007 May;27(21):5730–43.

cortical neurons. J Neurosci. 2000;20(10):3864–73.

Res Rev. 1997 Dec;25(3):312–34.

11;496(7444):224–8.

7.

[138] Buffalari DM, See RE. Inactivation of the bed nucleus of the stria terminalis in an animal model of relapse: effects on conditioned cue-induced reinstatement and its enhance‐ ment by yohimbine. Psychopharmacology (Berl). 2011 Jan;213(1):19–27.
