nestler (2001) nature reviews neuroscience 2, 110-128. molecular and genetic bases of nicotine...
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Molecular and Genetic Bases of Nicotine DependenceNoboru Hiroi, Ph.D.
Department of Psychiatry and Behavioral SciencesDepartment of Neuroscience
Albert Einstein College of Medicine
Smoking as DependenceConfession of a smoker
- He smoked 22-40 cigars per day.
- "To cease smoking is the easiest thing I ever did. I ought to know because I've done it a thousand times."
- “As an example to others, and not that I care for moderation myself, it has always been my rule never to smoke when asleep* and never to refrain when awake.” --70th birthday speech
- * “He always went to bed with a cigar in his mouth, and sometimes, mindful of my fire insurance, I went up and took it away, still burning, after he had fallen asleep." William Dean Howells.
Samuel Langhorne Clemens
-
“Nicotine is not addictive”
Nicotine as an addictive substance
- Smokers prefer nicotine-containing cigarettes to de- nicotinized cigarettes.
- Smokers experience withdrawal when switching to light cigarettes.
- Nicotine replacement alleviates withdrawal symptoms.
How easily would you develop dependence?
Nicotine
Heroin
Cocaine
Alcohol
Stimulants other than cocaine (d-amphetamine and methamphetamine)
Cannabis (marijuana, hashish, or both)
Anxiolytics/sedative and hypnotic drugs (secobarbital, diazepam, flurazepam, alprazolam, and triazolam)
Analgesics (morphine, propoxyphene, and codeine)
% of individuals with dependence among extra-medical users.
n=8,098, 15-54 years old. (Anthony et al., 1994)
32%
23%
17%
15%
11%
9%
9%
8%
How to define substance dependence (by features)Physiological Dependence• Tolerance • Withdrawal
Behavioral Dependence (Addiction)• Substance taken in larger amounts or over a longer period than
intended.• Persistent desire or unsuccessful attempts to control or
decrease use.• Great deal of time spent in activities to obtain substance.• Substance use significantly or completely takes place of social,
occupational, or recreational activities.• Continued use despite adverse consequences.(Three or more of the above within a 12 month period) DSM-IV-TR
Problems• All or none• Not mechanism based• Heterogeneous nature ignored• Not much translational value
Affiliative attachment Automaticity Behavioral choice-melioration Loss of control Cognitive enhancement Craving Cue exposure-associative processes Negative reinforcement Positive reinforcement Social and environmental goads Taste and sensory properties Tolerance
WISDM: Wisconsin Inventory of Smoking Dependence MotivesPiper et al (2004)
How to define substance dependence (by reasons)
Subjective reasons for smoking
Affiliative attachment
Automaticity
Behavioral choice-melioration
Loss of control
Cognitive enhancement
Craving
Cue exposure-associative processes
Negative reinforcement
Positive reinforcement Social and environmental goads
Taste and sensory properties
Tolerance
How to model elements of dependence in experimental animals
?
?
?
?
Memory tasks
?
Cue reactivity
Withdrawal and cue reactivity
Self-administration and cue control
?
?
?
Do you have an urge to smoke?
Drug-associated cues
Droungas, et al., 1995
Neutral cuesSmoking-related cuesUnpleasant cues
Nicotine
Rodent model of cue reactivity
CONDITIONING DAY: MORNING
Saline
CONDITIONING DAY: AFTERNOON
TEST DAY
Affiliative attachment
Automaticity
Behavioral choice-melioration
Loss of control
Cognitive enhancement
Craving
Cue exposure-associative processes
Negative reinforcement
Positive reinforcement Social and environmental goads
Taste and sensory properties
Tolerance
How to model elements of dependence in experimental animals
?
?
?
?
Memory tasks
?
Cue reactivity
Withdrawal and cue reactivity
Self-administration and cue control
?
?
?
Nicotine
Withdrawal Cue Aversion
Chronic nicotine intake
Mecamylamine
CONDITIONING DAY: MORNING
Precipitated withdrawal
Saline
No withdrawal
TEST DAY
N. Accumbens
DA
VTA
Nicotine Acts on the Mesolimbic Dopamine Pathway
Nicotinic receptorNicotine
Mesolimbic Dopamine Pathway
Caudate-Putamen
N. Accumbens
Frontal/cingulate cortex
VTA
Hiroi (2005)
Double Dissociation of Nicotine Cue Preference and Withdrawal-Cue Aversion
CPP
Pretest Test 1 Test 2 Test 3 Test 4 Test 5 Test 6 Test 7 Test 8 Test 9 Test 10Tim
e d
iffer
enc
e (
Nic
otin
e-p
aire
d -
sa
line
-pa
ired,
sec
)
-300
-200
-100
0
100
200
300
400 InsulaOrbitofrontal CortexSham Cortical control
Pretest Test 1 Test 2 Test 3 Test 4 Test 5 Test 6 Test 7 Test 8 Test 9 Test 10
Tim
e d
iffe
ren
ce (
Me
cam
yla
min
e-p
aire
d -
sa
line
-pa
ired
, se
c)
-400
-300
-200
-100
0
100
200
300
400InsulaOrbitofrontal CortexSham Cortical control
Scott and Hiroi (in preparation)
Nicotine Cue Apporach Withdrawal-Cue Aversion
N. Accumbens
DA
VTA
Nicotine Acts on the Mesolimbic Dopamine Pathway
Nicotinic receptorNicotine
Transcription Factor
Downstream genes
Second Messenger
Nicotine Dependence?
Dopamine
Hypothetical Intracellular Cascades of Addiction
?
Saline Nicotine Sal Nic Sal Nic Sal Nic
0.5 1 6 0.5 0.5 1 6 0.5 0.5 1 6 0.5 0.5 1 6 0.5
WT KO WT KO WT KO WT KO
Accumbens Caudate-Putamen Ventral tegmental area Substantia Nigra
FosB
DFosB
β-actin
Time (hr)
FosB/DFosB induction by nicotine along the mesolimbic dopamine pathway
N. Accumbens
Caudate-Putamen
VTASN
Zhu et al (2007) Human Molecular Genetics
Strategy
Mouse with deletion of single genes
Characterize behavioral phenotypes in rodentmodels of nicotine dependence
FosB KO mouse• Backcrossed to C57BL/6J mice for 7 generations
• Reduced locomotor habituation (Zhu et al., 2007)• Basally low blood corticosterone level (Zhu et al., 2007)
Normal in:• neurological functions (Gruda et al., 1996)• olfactory discrimination (Brown et al., 1996)• learning (Brown et al., 1996)• anatomy (Hiroi et al., 1997)• regulation of other Fos family proteins (Hiroi et al., 1999)• ethanol preference (Korkosz et al., 2004)• nicotine metabolism (Zhu et al., 2007)• novelty approach (Zhu et al., 2007)• anxiety-related behavior (Zhu et al., 2007)
Cue-induced Approach
Time difference = Time in nicotine side - time in saline side Zhu et al (2007) Human Molecular Genetics
Nicotine (base mg/kg)0.025 0.05 0.2 0.6 0.8 2
Tim
e di
ffere
nce
(sec
)
-400
-200
0
200
400 WT
FosB KO
*
Preference
Aversion
Transcription Factor
Downstream genes
cAMP
Nicotine Cue Approach
Dopamine
FosB
PKA
GC (soluble guanylate cyclase)
Nicotine Dependence?
Nitric Oxide (NO)
cGMP (cyclic guanosine monophosphate)
PKG (cGMP-dependent protein kinase)
PThr34P-Thr75
DARPP-32
Nicotine
Allen Brain Atlas (2009)
PKGII mRNA
Region-specific activation by nicotine of PKG activity
PFC NA CA AMY VTA SN
Per
cent
of c
ontr
ol
60
70
80
90
100
110
120
130
140
150
160
170
180SalineNicotine (0.2 mg/kg)
**
**
**
Scott et al., 2009
PKG-II KO mice
• Congenic C57BL/6 background• Reduced bone length• Defective intestinal water secretion• Enhanced anxiety• Increased ethanol consumption• Altered light-induced circadian clock
resetting
Maintenance of Nicotine Cue Approach
Pretest 1 2 3 4 5 6 7 8 9 10Tim
e di
ffere
nce
(sec
, nic
otin
e-pa
ired
- sa
line-
paire
d)
-400
-300
-200
-100
0
100
200
300
WTHTKO
**
** **
**
**
++++
+ +
#
PKGII KO
PKGII WT
Scott et al., 2009
Pretest Test 1 Test 2 Test 3 Test 4 Test 5 Test 6 Test 7 Test 8 Test 9 Test 10
Tim
e d
iffe
ren
ce (
me
cam
yla
min
e-p
aire
d -
sa
line
-pai
red,
se
c)
-500
-400
-300
-200
-100
0
100
200
WT HT KO
Withdrawal-Cue Aversion
Scott et al., 2009
GC
Long-term Nicotine Cue ApproachNot withdrawal-cue aversion
Nitric Oxide (NO)
cGMP
PKG
Nicotine
Nicotine
Long-term retention ofNicotine cue approachPKG-II
Acute Nicotine cue approachFosB
Translational Approach
Nicotine cue approachWithdrawal cue aversion
InsularOrbitofrtontal cortexStriatumAmygdala
FosBPKGDARPP-32MAOA
fosB 19q13.32Prkg2 4q13.1-q21.1DARPP-32 17q12Maoa Xp11.3
Albert Einstein College of MedicineLaboratory of Molecular PsychobiologyDepartment of Psychiatry and Behavioral SciencesDepartment of NeuroscienceNoboru Hiroi, Ph.D.Takehito Sawamura, M.D. Hongwen Zhu, M.D. , Ph.D. Soh Agatsuma, M.D., Ph.D.Go Suzuki, M.D.Takeshi Hiramoto, Ph.D.Kathryn Harper, M.Sc.Daniel Scott, M.Sc.Gina Kang, B.Sc.
Franz Hofmann, Ph.D. Jens Schlossmann, Ph.D. Robert Feil, Ph.D. Suzanne Feil, Ph.D.
Supported by: NIDA (R01DA13232 and R01DA024330), NARSAD, Transgenic Core/ Albert Einstein College of Medicine
The Rockefeller UniversityPaul Greengard, Ph.D.
Children’s Hospital/Harvard Medical SchoolMichael E. Greenberg, Ph.D.
University of Southern CaliforniaJean C. Shih, Ph.D.Kevin Chen, Ph.D.
University of Paris-SudIsabelle Seif
Technical University of Munich
ACKNOWLEDGEMENTS
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