Document Text Contents
Page 1
THE BASAL
GANGLIA III
Page 2
ADVANCES IN BEHAVIORAL BIOLOGY
Editorial Board
Jan Bures
Irwin Kopin
Bruce McEwen
James McGaugh
Institute of Physiology, Prague, Czechoslovakia
National Institute of Mental Health, Bethesda, Maryland
Rockefeller University, New York, New York
University of California, Irvine, California
Karl Prlbram
Jay Rosenblatt
Lawrence Welskrantz
Stanford University School of Medicine, Stanford, California
Rutgers University, Newark, New Jersey
University of Oxford, Oxford, England
Recent Volumes in this Series
Volume 27 THE BASAL GANGLIA: Structure and Function
Edited by John S. McKenzie, Robert E. Kemm,
and Lynette N. Wilcock
Volume 28 BRAIN PLASTICITY, LEARNING, AND MEMORY
Edited by B. E. Will, P. Schmitt, and
J. C. Dalrymple-Alford
Volume 29 ALZHEIMER'S AND PARKINSON'S DISEASES: Strategies
for Research and Development
Edited by Abraham Fisher, Israel Hanin, and Chaim Lachman
Volume 30 DYNAMICS OF CHOLINERGIC FUNCTION
Edited by Israel Hanin
Volume 31 TOBACCO SMOKING AND NICOTINE: A Neurobiological Approach
Edited by William R. Martin, Glen R. Van Loon, Edgar T. Iwamoto, and
Lay ten Davis
Volume 32 THE BASAL GANGLIA II: Structure and Function-Current Concepts
Edited by Malcolm B. Carpenter and A. Jayaraman
Volume 33 LECITHIN: Technological, Biological, and Therapeutic Aspects
Edited by Israel Hanin and G. Brian Ansell
Volume 34 ALTERATIONS IN THE NEURONAL CYTOSKELETON IN
ALZHEIMER' DISEASE
Edited by George P. Perry
Volume 35 MECHANISMS OF CEREBRAL HYPOXIA AND STROKE
Edited by George Somjen
Volume 36 NOVEL APPROACHES TO THE TREATMENT OF ALZHEIMER'S DISEASE
Edited by Edwin M. Meyer, James W. Simpkins, and Jyunji Yamamoto
Volume 37 KINDLING 4
Edited by Juhn A. Wada
Volume 38A BASIC, CLINICAL, AND THERAPEUTIC ASPECTS OF
ALZHEIMER'S AND PARKINSON'S DISEASES
Volume 1
Edited by Toshiharu Nagatsu, Abraham Fisher, and Mitsuo Yoshida
Volume 38B BASIC, CLINICAL, AND THERAPEUTIC ASPECTS OF
ALZHEIMER'S AND PARKINSON'S DISEASES
Volume 2
Edited by Toshiharu Nagatsu, Abraham Fisher, and Mitsuo Yoshida
Volume 39 THE BASAL GANGLIA '"
Edited by Giorgio Bernardi, Malcolm B. Carpenter, Gaetano Di Chiara,
Micaela Morelli, and Paolo Stanzione
A Continuation Order Plan is available for this series. A continuation order will bring delivery of each new
volume immediately upon publication. Volumes are billed only upon actual shipment. For further infonnation
please contact the publisher.
Page 374
REGULATION OF NT RECEPTORS AFTER CHRONIC TREATMENT WITH
TYPICAL AND ATYPICAL NEUROLEPTIC DRUGS
Laura Calza, Luciana Giardino, *Pier Vincenzo
Piazza and *Giuseppe Amato
Inst. Human Physiology, Medical School
University of Cagliari, Italy; *Inst. Human
Physiology, Medical School, University of Palermo
Italy
INTRODUCTION
Neurotensin is a tridecapeptide, which produces central
effects such as hypotension, hypothermia, muscle relaxation,
analgesia, antinociception, and reduces locomotor activity
(Nemeroff et al., 1977). In the recent years, several lines
of evidences indicated the great importance of the dopamine
(DA)- neurotensin (NT) interaction taking place mostly in the
mesolimbocortical areas, both at cell bodies (A1D DA-ergic
group located in the ventral tegmental area -VTA-) and nerve
terminal level (cerebral cortex, n. accumbens, olfactory
tubercle, ventral n. caudato-putamen) (Quirion, 1983;
Nemeroff, et al., 198~; Nemeroff, 1986). Immunocytochemical
data indicated the presence of NT-like immunoreactivity in DA
cell bodies in the VTA, including midline structures, with
only single example of coexistence in the substantia nigra
(Hokfelt et al., 198~). Anatomical data indicate that NT-like
immunoreactive fibres, nerve terminals and also cell bodies
are present in the basal ganglia (Kalivas, 198~; Sugimoto and
Mizuno, 1987; Uhl et aI, 1979). NT receptors are localized on
the DA-ergic cell bodies in the ventral mesencephalon (Young
and Kuhar, 1981). NT receptors are also reported to exist in
the basal ganglia of the rat, on DA nerve terminals,
intrinsic neurons and corticostriatal fibres (FIG. 1)
(Quirion et aI, 1985; Goedert et al., 198~).
Conventional antipsychotic drugs possess as their most
common side-effect the ability to produce extrapyramidal
symptoms. Certain of these agents produce a low incidence of
extrapyramidal side-effects and have been termed atypical
neuroleptics. A different action of typical and atypical
neuroleptics on the activity of A9 and A1D midbrain
dopaminergic neurons has been indicated as an argument for
site specificity in order to partially explain the different
clinical and behavioral properties of these two classes of
drugs (Chiodo and Bunney, 1983; White and Wang, 1983; Boris
and Diamond, 1983).
The Basa/ Ganglia Ill, Edited by G. Bernardi et a/.
Plenum Press, New York 379
Page 375
... DA neurons
• 0 A receptors
-e- NT neurons
121 NT receptors
o ather neurons
Nemeroff 1986 modified
P.'leios 1988 mDdified
Fig. 1. Schematic representation of the DA and NT receptor
localization on cell bodies and nerve terminals in
mesencephalic ascending pathways and related
structures.
In view of the different NT/DA interaction in the
mesostriatal and mesolimbocortical systems, we investigated
by means of quantitative receptor autoradiography the
modifications induced by the long-term treatment with
haloperidol low dosage, thioridazine, clozapine and
chlorpromazine on the DA2 and NT receptors in the substantia
nigra and ventral tegmental area, and the effect produced by
chronic clozapine on the DA2/NT receptor balance in the
target areas of the mesolimbocortical and mesostriatal
systems.
MATERIALS AND METHODS
Male Sprague Dawley rats, 200-250 gr body weight, were
used. The animals were housed in standard light and dark
conditions, with food pellets and water ad libitum. Groups of
10 rats were treated with the following drugs at the
indicated doses: haloperidol 0.5 mg/kg i.p., thioridazine 5
mg/kg in 0.2 ml saline orally, chlorpromazine 20 mg/Kg i.p.,
clozapine 20 mg/Kg i.p .. Saline 0.2 ml orally was used as
control of thioridazine treated rats, saline 0.2 ml i.p. as
control of other treatments. All the groups were treated for
21 days once a day. At the end of the treatments and after 3
days washout, the rats were sacrificed. Briefly, under
ketamine anaesthesia (10 mg/kg), the rats were perfused
through the ascending aorta (100 ml saline solution followed
by 100 ml paraformaldehyde 0.1% in PBS pH 7.~), then the
brains were quickly removed, frozen and sectioned in a
cryostat (Leitz Kriostat 1720, -25°C, 20 ~ thickness). 5
series of ~ consecutive sections (2 sections for each isoto-
pe, total and unspecific binding) were collected on gelatine
coated slides at the rostro-caudal level 1.2 anterior to the
bregma according to the Paxinos and Watson stereotaxic atlas
380
Page 747
Side effects
neuroleptic, 455, 471
Single-unit
activity, 249, 259, 541
recordings, 144, 231, 303, 463
Sistemic administration, 463
SKF38393, 249, 443, 591, 533
Slice preparations, 329, 275, 285
Somatic sensory cortex, 3
Somatostatin, 187, 199, 629
mRNA, 619
Somatotopic system, 495
Species susceptibility, 501, 511
related toxicity, 501, 511
Specific release, 407
Spine
density, 21
neck, 21
Spiny neurons, 187, 199
Spiperone binding, 379, 604
Spontaneous hyperactivity, 73
Spontaneous hypoactivity, 73
Staining intracellular, 551
Sterotypeis
apomorphine induced, 455
pilocarpine induced, 591
Stimulation
cerebellar, 144
corpus callosum, 293
cortical, 239
fornix, 231
nigral, 144, 239
putamen, 73
striatal, 73
Stored OA pool, 407
Striatal
cell recordings, 335
OA depletions, 335
intraocular transplants, 13
lesions, 433, 561
neurons, 222
stimulation, 73
transplants, 561
Striatonigral
GABAergic pathway, 481
pathway, 119, 249
Striato-pallidal pathway, 81
Striatum, 3, 13, 29, 49, 134, 371,
379, 433, 541
dorsal, 39, 425
ventral, 39, 425
Striosomal OA release, 363
Striosomal three dimensional
organization, 363
Striosomal separation, 363
Striosomes, 3, 39, 313
Subcortical demdntia, 703
Substance P (SP), 29, 39, 109, 119,
167, 187, 199, 629
ENK distribution, 39
immunoreactivity, 167
774
Substantia nigra, 29, 119, 134, 144,
187, 199, 259, 269, 275,
321, 481, 523, 541, 591
activity, 99
lesions, 541, 573
pars reticulata (SNR), 158, 417,
481
Subthalamic
lesions, 607
nucleus, 89, 99, 187, 199, 581,
607
nucleus afferents, 109
nucleus ventral striatal pathway,
89
output, 99
spontaneous activity, 99
Subthalamo-pallido pathway, 81, 99,
109, 607
Subthalamo-nigral pathway, 99, 481
Sulpiride, 321, 389, 455
Superior colliculus, 134
Super sensitivity , 455
c-fos related, 417
OA receptors, 443
01 and 02 blockade induced, 455
denervation, 391
receptors, 417
Susceptibility
species, 501, 511
Symmetrical synaptic contacts, 21,
119
Synaptic
contacts Ach-Sub P, 39
late excitation, 73
late inhibition, 73
plasticity, 231
systemic administration, 463
Tardive dyskinesia, 607
Terguride isomers, 391
Terminal
depolarization, 407
excitability, 249
receptor blockade, 249
receptor stimulation, 249
Tetrodotoxin (TTX)
insensitive release, 673
Thalamic
central complex, 177
complex, 607
GABAergic inhibition, 347
lesions, 607
Thalamotomy, 638
Thalamostriatal
pathways, 21
projection, 63
terminals, 167
Thalamo-cortico-caudal
pathways, 347
Thalamus, 177
Page 748
Therapeutic effects
clozapine, 471
neuroleptics, 471
Therapy
L-dopa, 646
Theta rhythm, 231
Thioridazine, 379, 471
Three dimensional striosomal
organization, 363
Time dependent rectification, 285
Toxicity
species related, 501, 511
Toxins
environmental, 717
Tracers
anterograde, 81, 177
retrograde, 81, 109, 134, 177
Tracing
anterograde, 81, 177
retrograde, 81, 134
Transient ischemia, 581
Transplant, 551
embryonic striatal intraocular,
13
Transplanted neostriatal neurons,
551
Transport
anterograde, 63, 144, 158
MPTP retrograde, 519
retrograde, 63, 144, 157
Treatment chronic, 379
Tremor pilocarpine induced, 591
Threshold action potential, 227
Transmitter release, 231
True blue (TB) 134
TTX-insensitive OA release, 363
TTX-sensitive OA release, 363
TTX resistant release stimulation,
347
Turning behaviour, 443
TWitch movements, 653
Typical neuroleptics, 379
Tyrosine hydroxylase (TH), 187,
199, 363, 519, 523
Ultrastructure, 187, 199
Uptake
dopamine, 501, 511
MPP+, 519
Vasoactive intestinal polypeptide
(VIP), 187, 199
Ventral tegmental area (VTA), 187,
199, 285, 523
Ventral
metabolic lesion, 581
striatum, 39, 89, 167, 231,
425, 471
striatum 02 receptors, 471
Ventromedial thalamic nucleus,
541
Vertical system, 495
Visual
discrimination, 743
dysfunctions, 654
processing, 743
spatial functions, 703
target, 646
Voltage clamp, 275, 285
Voltametric measurements OA, 49
WAIS scale, 697
Wearing-off, 723
Well water, 723
What to do, 495
Yawning, 604
Yohimbine, 389
775
THE BASAL
GANGLIA III
Page 2
ADVANCES IN BEHAVIORAL BIOLOGY
Editorial Board
Jan Bures
Irwin Kopin
Bruce McEwen
James McGaugh
Institute of Physiology, Prague, Czechoslovakia
National Institute of Mental Health, Bethesda, Maryland
Rockefeller University, New York, New York
University of California, Irvine, California
Karl Prlbram
Jay Rosenblatt
Lawrence Welskrantz
Stanford University School of Medicine, Stanford, California
Rutgers University, Newark, New Jersey
University of Oxford, Oxford, England
Recent Volumes in this Series
Volume 27 THE BASAL GANGLIA: Structure and Function
Edited by John S. McKenzie, Robert E. Kemm,
and Lynette N. Wilcock
Volume 28 BRAIN PLASTICITY, LEARNING, AND MEMORY
Edited by B. E. Will, P. Schmitt, and
J. C. Dalrymple-Alford
Volume 29 ALZHEIMER'S AND PARKINSON'S DISEASES: Strategies
for Research and Development
Edited by Abraham Fisher, Israel Hanin, and Chaim Lachman
Volume 30 DYNAMICS OF CHOLINERGIC FUNCTION
Edited by Israel Hanin
Volume 31 TOBACCO SMOKING AND NICOTINE: A Neurobiological Approach
Edited by William R. Martin, Glen R. Van Loon, Edgar T. Iwamoto, and
Lay ten Davis
Volume 32 THE BASAL GANGLIA II: Structure and Function-Current Concepts
Edited by Malcolm B. Carpenter and A. Jayaraman
Volume 33 LECITHIN: Technological, Biological, and Therapeutic Aspects
Edited by Israel Hanin and G. Brian Ansell
Volume 34 ALTERATIONS IN THE NEURONAL CYTOSKELETON IN
ALZHEIMER' DISEASE
Edited by George P. Perry
Volume 35 MECHANISMS OF CEREBRAL HYPOXIA AND STROKE
Edited by George Somjen
Volume 36 NOVEL APPROACHES TO THE TREATMENT OF ALZHEIMER'S DISEASE
Edited by Edwin M. Meyer, James W. Simpkins, and Jyunji Yamamoto
Volume 37 KINDLING 4
Edited by Juhn A. Wada
Volume 38A BASIC, CLINICAL, AND THERAPEUTIC ASPECTS OF
ALZHEIMER'S AND PARKINSON'S DISEASES
Volume 1
Edited by Toshiharu Nagatsu, Abraham Fisher, and Mitsuo Yoshida
Volume 38B BASIC, CLINICAL, AND THERAPEUTIC ASPECTS OF
ALZHEIMER'S AND PARKINSON'S DISEASES
Volume 2
Edited by Toshiharu Nagatsu, Abraham Fisher, and Mitsuo Yoshida
Volume 39 THE BASAL GANGLIA '"
Edited by Giorgio Bernardi, Malcolm B. Carpenter, Gaetano Di Chiara,
Micaela Morelli, and Paolo Stanzione
A Continuation Order Plan is available for this series. A continuation order will bring delivery of each new
volume immediately upon publication. Volumes are billed only upon actual shipment. For further infonnation
please contact the publisher.
Page 374
REGULATION OF NT RECEPTORS AFTER CHRONIC TREATMENT WITH
TYPICAL AND ATYPICAL NEUROLEPTIC DRUGS
Laura Calza, Luciana Giardino, *Pier Vincenzo
Piazza and *Giuseppe Amato
Inst. Human Physiology, Medical School
University of Cagliari, Italy; *Inst. Human
Physiology, Medical School, University of Palermo
Italy
INTRODUCTION
Neurotensin is a tridecapeptide, which produces central
effects such as hypotension, hypothermia, muscle relaxation,
analgesia, antinociception, and reduces locomotor activity
(Nemeroff et al., 1977). In the recent years, several lines
of evidences indicated the great importance of the dopamine
(DA)- neurotensin (NT) interaction taking place mostly in the
mesolimbocortical areas, both at cell bodies (A1D DA-ergic
group located in the ventral tegmental area -VTA-) and nerve
terminal level (cerebral cortex, n. accumbens, olfactory
tubercle, ventral n. caudato-putamen) (Quirion, 1983;
Nemeroff, et al., 198~; Nemeroff, 1986). Immunocytochemical
data indicated the presence of NT-like immunoreactivity in DA
cell bodies in the VTA, including midline structures, with
only single example of coexistence in the substantia nigra
(Hokfelt et al., 198~). Anatomical data indicate that NT-like
immunoreactive fibres, nerve terminals and also cell bodies
are present in the basal ganglia (Kalivas, 198~; Sugimoto and
Mizuno, 1987; Uhl et aI, 1979). NT receptors are localized on
the DA-ergic cell bodies in the ventral mesencephalon (Young
and Kuhar, 1981). NT receptors are also reported to exist in
the basal ganglia of the rat, on DA nerve terminals,
intrinsic neurons and corticostriatal fibres (FIG. 1)
(Quirion et aI, 1985; Goedert et al., 198~).
Conventional antipsychotic drugs possess as their most
common side-effect the ability to produce extrapyramidal
symptoms. Certain of these agents produce a low incidence of
extrapyramidal side-effects and have been termed atypical
neuroleptics. A different action of typical and atypical
neuroleptics on the activity of A9 and A1D midbrain
dopaminergic neurons has been indicated as an argument for
site specificity in order to partially explain the different
clinical and behavioral properties of these two classes of
drugs (Chiodo and Bunney, 1983; White and Wang, 1983; Boris
and Diamond, 1983).
The Basa/ Ganglia Ill, Edited by G. Bernardi et a/.
Plenum Press, New York 379
Page 375
... DA neurons
• 0 A receptors
-e- NT neurons
121 NT receptors
o ather neurons
Nemeroff 1986 modified
P.'leios 1988 mDdified
Fig. 1. Schematic representation of the DA and NT receptor
localization on cell bodies and nerve terminals in
mesencephalic ascending pathways and related
structures.
In view of the different NT/DA interaction in the
mesostriatal and mesolimbocortical systems, we investigated
by means of quantitative receptor autoradiography the
modifications induced by the long-term treatment with
haloperidol low dosage, thioridazine, clozapine and
chlorpromazine on the DA2 and NT receptors in the substantia
nigra and ventral tegmental area, and the effect produced by
chronic clozapine on the DA2/NT receptor balance in the
target areas of the mesolimbocortical and mesostriatal
systems.
MATERIALS AND METHODS
Male Sprague Dawley rats, 200-250 gr body weight, were
used. The animals were housed in standard light and dark
conditions, with food pellets and water ad libitum. Groups of
10 rats were treated with the following drugs at the
indicated doses: haloperidol 0.5 mg/kg i.p., thioridazine 5
mg/kg in 0.2 ml saline orally, chlorpromazine 20 mg/Kg i.p.,
clozapine 20 mg/Kg i.p .. Saline 0.2 ml orally was used as
control of thioridazine treated rats, saline 0.2 ml i.p. as
control of other treatments. All the groups were treated for
21 days once a day. At the end of the treatments and after 3
days washout, the rats were sacrificed. Briefly, under
ketamine anaesthesia (10 mg/kg), the rats were perfused
through the ascending aorta (100 ml saline solution followed
by 100 ml paraformaldehyde 0.1% in PBS pH 7.~), then the
brains were quickly removed, frozen and sectioned in a
cryostat (Leitz Kriostat 1720, -25°C, 20 ~ thickness). 5
series of ~ consecutive sections (2 sections for each isoto-
pe, total and unspecific binding) were collected on gelatine
coated slides at the rostro-caudal level 1.2 anterior to the
bregma according to the Paxinos and Watson stereotaxic atlas
380
Page 747
Side effects
neuroleptic, 455, 471
Single-unit
activity, 249, 259, 541
recordings, 144, 231, 303, 463
Sistemic administration, 463
SKF38393, 249, 443, 591, 533
Slice preparations, 329, 275, 285
Somatic sensory cortex, 3
Somatostatin, 187, 199, 629
mRNA, 619
Somatotopic system, 495
Species susceptibility, 501, 511
related toxicity, 501, 511
Specific release, 407
Spine
density, 21
neck, 21
Spiny neurons, 187, 199
Spiperone binding, 379, 604
Spontaneous hyperactivity, 73
Spontaneous hypoactivity, 73
Staining intracellular, 551
Sterotypeis
apomorphine induced, 455
pilocarpine induced, 591
Stimulation
cerebellar, 144
corpus callosum, 293
cortical, 239
fornix, 231
nigral, 144, 239
putamen, 73
striatal, 73
Stored OA pool, 407
Striatal
cell recordings, 335
OA depletions, 335
intraocular transplants, 13
lesions, 433, 561
neurons, 222
stimulation, 73
transplants, 561
Striatonigral
GABAergic pathway, 481
pathway, 119, 249
Striato-pallidal pathway, 81
Striatum, 3, 13, 29, 49, 134, 371,
379, 433, 541
dorsal, 39, 425
ventral, 39, 425
Striosomal OA release, 363
Striosomal three dimensional
organization, 363
Striosomal separation, 363
Striosomes, 3, 39, 313
Subcortical demdntia, 703
Substance P (SP), 29, 39, 109, 119,
167, 187, 199, 629
ENK distribution, 39
immunoreactivity, 167
774
Substantia nigra, 29, 119, 134, 144,
187, 199, 259, 269, 275,
321, 481, 523, 541, 591
activity, 99
lesions, 541, 573
pars reticulata (SNR), 158, 417,
481
Subthalamic
lesions, 607
nucleus, 89, 99, 187, 199, 581,
607
nucleus afferents, 109
nucleus ventral striatal pathway,
89
output, 99
spontaneous activity, 99
Subthalamo-pallido pathway, 81, 99,
109, 607
Subthalamo-nigral pathway, 99, 481
Sulpiride, 321, 389, 455
Superior colliculus, 134
Super sensitivity , 455
c-fos related, 417
OA receptors, 443
01 and 02 blockade induced, 455
denervation, 391
receptors, 417
Susceptibility
species, 501, 511
Symmetrical synaptic contacts, 21,
119
Synaptic
contacts Ach-Sub P, 39
late excitation, 73
late inhibition, 73
plasticity, 231
systemic administration, 463
Tardive dyskinesia, 607
Terguride isomers, 391
Terminal
depolarization, 407
excitability, 249
receptor blockade, 249
receptor stimulation, 249
Tetrodotoxin (TTX)
insensitive release, 673
Thalamic
central complex, 177
complex, 607
GABAergic inhibition, 347
lesions, 607
Thalamotomy, 638
Thalamostriatal
pathways, 21
projection, 63
terminals, 167
Thalamo-cortico-caudal
pathways, 347
Thalamus, 177
Page 748
Therapeutic effects
clozapine, 471
neuroleptics, 471
Therapy
L-dopa, 646
Theta rhythm, 231
Thioridazine, 379, 471
Three dimensional striosomal
organization, 363
Time dependent rectification, 285
Toxicity
species related, 501, 511
Toxins
environmental, 717
Tracers
anterograde, 81, 177
retrograde, 81, 109, 134, 177
Tracing
anterograde, 81, 177
retrograde, 81, 134
Transient ischemia, 581
Transplant, 551
embryonic striatal intraocular,
13
Transplanted neostriatal neurons,
551
Transport
anterograde, 63, 144, 158
MPTP retrograde, 519
retrograde, 63, 144, 157
Treatment chronic, 379
Tremor pilocarpine induced, 591
Threshold action potential, 227
Transmitter release, 231
True blue (TB) 134
TTX-insensitive OA release, 363
TTX-sensitive OA release, 363
TTX resistant release stimulation,
347
Turning behaviour, 443
TWitch movements, 653
Typical neuroleptics, 379
Tyrosine hydroxylase (TH), 187,
199, 363, 519, 523
Ultrastructure, 187, 199
Uptake
dopamine, 501, 511
MPP+, 519
Vasoactive intestinal polypeptide
(VIP), 187, 199
Ventral tegmental area (VTA), 187,
199, 285, 523
Ventral
metabolic lesion, 581
striatum, 39, 89, 167, 231,
425, 471
striatum 02 receptors, 471
Ventromedial thalamic nucleus,
541
Vertical system, 495
Visual
discrimination, 743
dysfunctions, 654
processing, 743
spatial functions, 703
target, 646
Voltage clamp, 275, 285
Voltametric measurements OA, 49
WAIS scale, 697
Wearing-off, 723
Well water, 723
What to do, 495
Yawning, 604
Yohimbine, 389
775
