Journal of Neurochemistry
Lippincott—Raven Publishers, Philadelphia
© 1997 International Society for Neurochemistry
Angiotensin-Converting Enzyme Modulates Dopamine
Turnover in the Striatum
Trisha A. Jenkins, *Frederick A. 0. Mendelsohn, and *Sjew Yeen Chai
Department of Medicine, University of Melbourne, Austin and Repatriation Medical Centre, Austin Campus, Heidelberg;
and * Howard Florey Institute of Experimental Physiology and Medicine,
University of Melbourne, Parkville, Victoria, Australia
Abstract: The effect of chronic inhibition of the angioten-
sin-converting enzyme on dopamine content and release
in the striatum was investigated using in vivo microdialy-
sis in awake, freely moving rats. Rats were treated for 1
week with the angiotensin-converting enzyme inhibitor
perindopril (1 mg/kg) via the drinking water, whereas the
controls were given water alone. One week after perindo-
pril treatment, striatal dopamine dialysate levels in the
treated group were markedly elevated compared with
control values: control, 233 ± 43 pg/mI; perindopril, 635
±53 pg/mI (p < 0.001). These results were confirmed
by a complementary study in which dopamine content
was measured in striatal extracts (3.5 ±0.4 ~zgof dopa-
mine/g of tissue for controls compared with 9.2 ± 2.4
~zgof dopamine/g of tissue for the treated group; p
<0.05). In the rats that were dialyzed, angiotensin-con-
verting enzyme levels in the striatum were decreased by
50% after perindopril treatment. Levels of dopamine D1
and D2 receptors and of preprotachykinin and tyrosine
hydroxylase mRNA5 were unchanged after angiotensin-
converting enzyme inhibition. A small, but significant, in-
crease was detected in striatal preproenkephalin mRNA
levels in the angiotensin-converting enzyme inhibitor-
treated group. These results indicate that peripherally ad-
ministered angiotensin-converting enzyme inhibitors pen-
etrate the blood—brain barrier when given chronically and
modulate extracellular dopamine and striatal neuropep-
tide levels. Key Words: Angiotensin-converting en-
zyme—Angiotensin-converting enzyme inhibitor—Do-
pamine—Perindopril—Striatum—Enkephalin.
J. Neurochem. 68, 1304—1311 (1997).
Angiotensin-converting enzyme (ACE) inhibitors
are widely used clinically in the treatment of hyperten-
sion and heart failure. Reports of mood elevation (Zu-
benko and Nixon, 1984), a feeling of well-being, and
better quality of life (Croog et al., 1986) are common;
these effects are considered to be the result of ACE
inhibitors crossing the blood—brain barrier to have cen-
tral effects on brain ACE (Beto and Bansal, 1992). In
rats we have shown that ACE inhibitors not only block
ACE in the circumventricular organs (Sakaguchi et al.,
1304
1987), but also cross the blood—brain barrier to block
ACE in the striatum (Chai et al. 1992).
A high level of ACE activity exists within the rat
basal ganglia, including the substantia nigra, globus
pallidus, and caudate putamen (Chai et a!., 1987b). We
have previously demonstrated with selective chemical
lesions of the striatum and substantia nigra that ACE
in the basal ganglia is associated with neurons in the
striatum and their striatonigral and striatopallidal pro-
jections, but not with the ascending nigrostriatal dopa-
minergic system (Chai et al., 1987a). Although both
angiotensin II (Ang II) immunoreactivity and the AT1
receptors are found in the caudate putamen of rats
(Harding et al., 1981; Simonnet and Vincent, 1982;
Lind et al., 1985), in the human brain, the AT1 recep-
tors occur in association with the nigrostriatal dopa-
minergic system (Allen et al., 1991). Physiological
studies with slices in vitro (Simonnet and Vincent,
1982; Dwoskin et al., 1992), in vivo microdialysis
(Mendelsohn et al., 1993), and behavioral models
(Jenkins et al., 1995) demonstrate that these AT1 re-
ceptors are functionally active.
It has been proposed that in addition to its well-
known action in the conversion of angiotensin I to Ang
II and in the breakdown of bradykinin, ACE in the
brain may have other roles. ACE is capable of hy-
drolyzing several neuropeptides, including [Met I and -
[Leu I -enkephalin, substance P. neurotensin, dynor-
phin,6-Phe7
and the enkephalin
(Skidgel precursor
et al., 1987), [Met] -enkephalin-
that may play im-
Arg
portant roles in basal ganglia function. Inhibition of
Received May 21, 1996; revised manuscript received September
25, 1996; accepted November 12, 1996.
Address correspondence and reprint requests to Dr. T. A. Jenkins
at Department of Medicine, University of Melbourne, Austin and
Repatriation Medical Centre, Austin Campus, Studley Road, Heidel-
berg, Victoria, 3084, Australia.
Abbreviations used: ACE, angiotensin-converting enzyme; Ang
II, angiotensin II; DOPAC, 3,4-dihydroxyphenylacetic acid; GFAP,
glial fibrillary acidic protein; HVA, homovanillic acid; SSC, saline—
sodium citrate.
 ACE INHIBITORS AND DOPAMINE
metabolism of these neuropeptides may contribute
partly to the central effects attributed to ACE inhibi-
tors.
Considering the high levels of ACE within the stria-
turn and the abundance of neuropeptides in the caudate
putarnen known to be influenced by ACE, this study
investigated the effect of chronic ACE inhibition on
levels of dopamine within the striatum and the concur-
rent changes in neuropeptide synthesis.
MATERIALS AND METHODS
Study 1: effect of chronic ACE inhibition on
striatal dopamine release and striatal ACE and
neuropeptide mRNA levels
In vivo microdialysis. Fourteen male Sprague—Dawley
rats (weighing 200—250 g) were anesthetized with a metho-
hexital sodium (Brietal)/amobarbital sodium (Amytal) mix
(Eli Lilly, Australia; 1.5 mg/kg, i.p.) and placed in a stereo-
taxic frame. A guide cannula, with a dummy probe (CMA
Microdialysis, Sweden), was implanted into the right stria-
turn (0.2 mm posterior to the bregma, 3.0 mm right of the
sagittal suture, and 4.0 mm below the dura). Animals were
allowed to recoverfor at least 7 days. Experimental protocols
were approved by the Austin Hospital Animal Ethics Com-
mittee and meet the guidelines of the National Health and
Medical Research Council, Commonwealth Scientific and
Industrial Research Organization, and Australian Agricul-
tural Council Joint Committee.
On the morning of the experiment, the dummy probe was
replaced with a microdialysis probe (0.24 mm in diameter,
3 mm long; CMA Microdialysis, Sweden), through which
an artificial CSF solution containing 133 mM Na~,3.0 mM
K, 2.0 mM Ca2~,0.4 mMMg2~,132 mMCl~,0.5mM
H 2-, 3.6 mM glucose,
3.3 mM
2P04,
and 3 mMureaHC03,
was perfused
0.4 mMatSO4
2 ~il/min. Experimental
collections began 1 h after probe implantation, to allow base-
line stabilization.
Six 20-mm dialysate samples were collected and analyzed
from each rat to determine baseline levels of amines. Rats
were then randomly divided into two groups: One group
received the ACE inhibitor perindopril (1 mg/kg in the
drinking water) for 7 days, and the other received water
alone. At the end of the week dialysate samples were again
collected from the striatum and analyzed.
HPLC analysis of arnines and metabolites. Samples were
collected every 20 mm into vials containing 10 pl of 0.1 M
perchioric acid (AR grade; Mallinckrodt U.S.A.) and within
5 mm injected onto an HPLC system, composed of a Bioana-
lytical Systems (U.S.A.) model PM-80 solvent delivery sys-
tem, a Linear RYYT chart recorder (Bioanalytical Systems),
a CMA 200 refrigerated microsampler (CMA Microdialysis,
Sweden), an Altech 15-cm Econosphere C
18 5-~tm(particle
size) reverse-phase column, a Bioanalytical Systems model
LC-4C electrochemical detector, and an RE-4 flow cell with
a dual glassy carbon working electrode. The applied voltage
of the detector was set at + 0.65 V relative to the Ag/AgC1
reference electrode. The mobile phase consisted of 50 mM
sodiumphosphate, 20 mM sodium citrate, 2 mM heptanesul-
fonic acid, 0.1 mM EDTA, and 15% methanol, pH 3.0. The
detection limits of this system, defined as masses of stan-
dards producing peak heights of double the baseline noise,
1305
were 1.5 pg for dopamine, 23 pg for 3,4-dihydroxyphenyl-
acetic acid (DOPAC), and 12 pg for homovanillic acid
(HVA) (Sigma, U.S.A.).
In vitro histological analysis. After the in vivo microdialy-
sis procedure, the animals were anesthetized with fluothane
inhalation and decapitated, trunk blood was collected, and
the brain was excised and frozen at —40°C.Tissue sections
were cut 10 and 20 jzm thick on a Microm HM5O5E cryostat
at —20°Cand thawed onto coated glass slides. For the autora-
diographic technique, 20-tim sections were collected onto
gelatin-coated slides, dehydrated overnight at 4°Cunder re-
duced pressure, and stored at —80°C with desiccant until
required. For in situ hybridization and immunohistochemical
studies, 10-gm sections were collected onto poly-L-lysine-
coated slides, stored in the chilled cryostat after thaw mount-
ing, and then kept until needed at —80°C.Alternate 20-~tm
sections were Nissl-stained with thionin to verify microdialy-
sis probe localization. The collecting area of the dialysis
probe was always located within the middle of the caudate
putamen.
In vitro autoradiography:ACE. Methods for in vitro auto-
radiographic localization of ACE with 124135 1A, a tyrosyl
derivative of the ACE inhibitor lisinopril, which binds to
the catalytic site of ACE, have been previously described
(Mendelsohn et al., l984a,b). In brief, 20-~rmsections were
preincubated for 15 mm in 10 mM sodium phosphate buffer
(pH 7.4) containing 150 mM NaCl and 0.2% bovine serum
albumin before a 1-h incubation in a fresh volume of the
same buffer containing ~-~5X l0~cpmlml (100 pM) l25J
351A, iodinated by the chloramine T method (Hunter and
Greenwood, 1962). Nonspecific binding was measured in
the presence of 1 mM Na2EDTA. After incubation, sections
were washed four times, for 1 mm each, in ice-cold buffer
without bovine serum albumin, to remove nonspecifically
bound radioligand. Slides were dried, loaded into x-ray cas-
settes with a series of appropriate radioactivity standards,
exposed to Kodak-Ektascan EB-1 x-ray film (Eastman Ko-
dak Co., U.S.A.) at room temperature for 3 days, and then
processed in an RPX-OMAT 3H}SCH automatic 23390
developer.
binding was
performed receptors.(Savasta
DopamineasD1described [ et al., 1986). Slide-
mounted sections (20 tim) were preincubated for 15 mm at
room temperature in 50 mM Tris-HC1 buffer containing 20
mM NaCl, 5 mM KC1, 1 mM MgC1
2, and 2 mM CaCI2 (pH
7.4) before a 1-h incubation in a fresh
3H]SCH volume
23390 (71.3
ofCi/mmol;
the same
buffer
NEN Du containing ‘-~lnMCO,
Pont, Denver, [ U.S.A.). Nonspecific binding
was measured in the presence of 1 ~tM SCH 23390 (Research
Biochemicals International, U.S.A.). After incubation, sec-
tions were dipped briefly in ice-cold buffer before being
washed twice, for 10 mm each, in buffer and subsequently
rinsed in ice-cold water. The sections were then exposed to
Hyperfilm-3H (Amersham, Sweden) for 5 days and then
developed manually in Kodak Dl9 developer (Eastman Ko-
dak Co., U.S.A.).
Dopamine D
2 receptors. (Black
3H]sulpiride Dopamine
and Crossman,
D2 receptors1992).
were la-
In
brief, using
beled 20-,um[ sections were preincubated for 15 mm in 50
mM Trms-HC1 buffer with 120 mM NaCl (pH 7.7) before a
30-mm incubation in a fresh volume of the same buffer
containing 1 nM [3H] sulpiride (69 Ci/mmol; NEN Du Pont,
U.S.A.). Nonspecific binding was measured in the presence
of 1 jiM sulpiride (Research Biochemicals International,
J. Neurochem., Vol. 68, No. 3, 1997
1306 T. A. JENKINS ET AL.
U.S.A.). After incubation, sections were washed in fresh ice-
cold buffer, twice for 10 s each, before being rinsed in water
at 0°Cto remove nonspecifically bound radioligand. The
3H for 21 days and
sections were then exposed to Hyperfilm-
then developed manually in Kodak D19 developer.
In situ hybridization histochemistry. Levels of mRNA
were measured using in situ hybridization with radiolabeled
oligonucleotides. The oligonucleotides used were sequences
from rat preprotachykinin (145—192) (Krause et al., 1987),
rat preproenkephalin (322—361)(Howells et al., 1984), and
rat tyrosine hydroxylase (477—493) (Grima et al., 1985).
The in situ hybridization method was adapted from that of
Schalling et al. (1988).
In brief, all oligonucleotides were 3 ‘-end-labeled with 35~~
dATP (NEN Du Pont, U.S.A.) using terminal d-transferase.
Sections (10 jim) were hybridized at 42°C for 18 h in a
solution containing 1 X lO7cpm/ml probe (—~5X 105/jil).
The hybridization cocktail consisted of 50% formamide, 4
>< saline—sodium citrate (SSC; 0.6 M NaC1 and 0.06 M
sodium citrate), 1 x Denhardt’ s solution (2% Ficoll, 2%
bovine serum albumin, and 2% polyvinylpyrrolidone), 2%
sarcosyl, 0.2 M Na
2HPO4 buffer, 10% dextran sulfate, her-
ring sperm DNA (50 jil/ml), and 5 M dithiothreitol (40 jil/
ml). After hybridization, sections were washed four times
in 1 x SSC for 30 mm at 55°C,rinsed in sterile water, and
dehydrated through a succession of graded ethanol solutions.
Sections from the in situ hybridization histochemical proce-
dure were apposed to Hyperfilm-/3max (Amersham, Swe-
den) for 14—21 days before being hand-developed using
Kodak D19 developer.
Analysis of the in vitro autoradiography and in situ hy-
bridization histochemical data. Optical density on the films
was quantified using an MCID microcomputer 3H, or 35S imaging
radioactivity
sys-
tem
standards
(Imaging
were
Research,
fitted to Canada)
a calibration
.1251, curve, and the optical
density of sections was mapped into dpm of radioligand per
square millimeter for 125J and 35S compounds (Sexton et al.,
1986) and into nanocuries per milligram of tissue for the
tritiated compounds (Amersham, Sweden). Differences be-
tween control and ACE inhibitor-treated data were analyzed
using an unpaired t test.
Glialfibrillary acidic protein (GFAP) immunohistochem-
istry. Astrocytes in the area around the implanted microdialy-
sis probe were stained by the indirect immunoperoxidase
method with antibody to GFAP (rabbit anti-cow; DAKO,
Denmark). The working GFAP dilution was 1:1,000, with
an overnight incubation.
Study 2: effect of chronic ACE inhibition on
striatal dopamine content
Male Sprague—Dawley rats (weighing 200—250 g) re-
ceived either perindopril (1 mg/kg in drinking water) or
water for 1 week (n = 6 for each group). At the end of the
treatment period animals were killed. The brain was re-
moved, and the striatum and frontal cortex were dissected,
frozen in isopentane chilled with dry ice, and stored at
—80°C.
Dopamine extraction. Tissues were thawed, weighed, and
sonicated at 60 W, 40% pulsed power (Branson Sonifier
Cell Disrupter B 15) for 15 s in 0.1 M perchloric acid with
0.1 mM Na
2EDTA (Murai et a!., 1988) using 0.04 g/100
j.tl for striatum and 0.12 g/100 jil for frontal cortex. The
homogenate was centrifuged at 1,500 g in a Beckman GPR
(U.S.A.) centrifuge for 15 mm at 4°C,the supernatant was
J. Neurochem., Vol. 68, No. 3, 1997
removed and further centrifuged at 12,000 rpm for 15 mm
in a Beckman Microcentrifuge, and the final supernatant was
collected.
The supernatant was injected onto the HPLC system
described above and analyzed for dopamine, DOPAC,
and HVA.
RESULTS
Study 1: effect of chronic ACE inhibition on
striatal dopamine release and striatal ACE and
neuropeptide mRNA levels
Dopamine release. After treatment with the ACE
inhibitor perindopril, the levels of extracellular dopa-
mine increased in the treatment group to —~2.5times
that of the control. Pretreatment baseline striatal dopa-
mine levels between the two groups were not signifi-
cantly different (Table 1).
Levels of the metabolites DOPAC and HVA did not
change significantly after ACE inhibition (Table 1).
In vitro autoradiography. Striatal ACE levels were
50% lower in the group of rats treated with perindopril,
compared with controls (Fig. 1A and B). No discern-
ible differences were observed in levels of doparnine
D1 and D2 receptors in the rat striatum after chronic
ACE inhibition (Fig. iC—F).
In situ hybridization histochemistry. After chronic
treatment with the ACE inhibitor perindopril, a small,
but significant, 15% increase in the levels of striatal
preproenkephalin mRNA was observed, compared
with controls (Fig. 2A and B). Levels of preprotach-
ykinin mRNA in the striatum (Fig. 2C and D) and
tyrosine hydroxylase mRNA levels in the substantia
nigra (Fig. 2E and F) did not change after ACE inhibi-
tor treatment.
GFAP. There were no discernible differences in the
amount of gliosis in the area of collection around the
microdialysis probe between the two treatment groups.
Study 2: effect of chronic ACE inhibition on
striatal dopamine content
Striatal dopamine levels. Chronic treatment with the
ACE inhibitor perindopril increased striatal dopamine
nearly threefold compared with the control group.
Again, we observed no significant differences in the
levels of the metabolites DOPAC and HVA, although,
as before, levels appeared higher in the treated animals
(Table 2).
Frontal cortex dopamine levels. Dopamine,
DOPAC, and HVA levels in the frontal cortex were
unchanged after chronic ACE inhibitor treatment (Ta-
ble 2).
DISCUSSION
A 1-week oral treatment with the ACE inhibitor
perindopril increased the content and release of dopa-
mine in the rat striatum to approximately three times
that of controls. No differences were seen in the levels
 ACE INHIBITORS AND DOPAMINE 1307

TABLE 1. Effect of chronic ACE inhibition on extracellular dopamine levels

in the rat striatum

Pretreatment Posttreatment
ACE ACE
Control inhibitor-treated Control inhibitor-treated p value

Dopamine (pg/ml) 443 ±123 615 ±75 233 ±43 635 ±53 p < 0.001
DOPAC (ng/ml) 189 ±33 259 ±32 212 ±42 307 ±57 p = 0.07
HVA (ng/ml) 26 ±5.5 35 ±4.2 34 ±7.0 48 ±9.0 p = 0.15

Levels of dopamine, DOPAC, and HVA were measured by in vivo microdialysis in the striatum before
and after 1 week of ACE inhibition and compared with values for untreated controls. Data are mean
±SEM values (n = 7).

of the major dopamine metabolites DOPAC and HVA
after treatment.
It has previously been reported that a single dose of
the ACE inhibitor ceronapril (10 jig/kg i.p.) increased
the level of dopamine in the amygdala and that of
HVA in the septum and decreased the level of DOPAC
in the nucleus accumbens. No significant changes in
dopamine levels were detected in the frontal cortex,
entorhinal cortex, olfactory tubercle, hippocampus, or
striatum (Barnes et al., 1992).
We have also shown in this study that after 1 week
of oral treatment, perindopril is able to cross the
blood—brain barrier to block ACE in the striatum. This
is in agreement with our previous study in which rats
administered a single dose of perindopril by gavage at
doses of 4—16 mg/kg showed decreased ACE levels
in the caudate putamen (Sakaguchi et al., 1988). It is
interesting that a previous study using chronic doses
of ACE inhibitors reported differential degree of inhi-
bition in the various brain regions. A i-week oral treat-
ment with the ACE inhibitors ramipril and Hoe 288
(10 mg/kg) resulted in marked inhibition of ACE ac-
tivity in the cortex, hypothalamus, and brainstem, but
both inhibitors failed to block striatal ACE (Gohlke et
al., 1989).
The classical role of ACE is the conversion of inac-
tive angiotensin I to active Ang II by removal of C-
terminal dipeptide and the breakdown of bradykinin.

1251-351A
FIG.
binding Levels
1. (in of (A) ACE
dpm/mm2), assessed Dby
(C) dopamine
3HISCH23390 binding (in1 nCi/mg
receptors
of
tissue),
assessedand by (E)
[ dopamine D
3H]sulpiridebinding (in nCi/mg of tissue)
2 receptors in the
assessed
caudate
by [ putamen in control and ACE inhibitor-
treated (1 week of perindopril, 1 mg/kg via drinking
water) animals. Data are mean ±SE (bars) values
offour measurements from each rat (n = 7 rats). *p
< 0.05. Also shown are representative sections of
(B) ACE, (D) dopamine D
1 receptor binding, and
(F) dopamine D2 receptor binding in the caudate
putamen of a control animal. White represents high
levels of binding.

J. Neurochem., Vol. 68, No. 3, 1997

1308 T. A. JENKINS ET AL.

FIG. 2. Levels of (A) preproenkephalin mRNA (in

2) in the striatum, (C) preprotachykinin
dpm/mm
mRNA (in dpm/mm2) in the striatum, and (E) tyrosine
hydroxylase mRNA (in dpm/mm2) in the substantia
nigra assessed by in situ hybridization in control and
ACE inhibitor-treated (1 week of perindopril, 1 mg/
kg via drinking water) animals. Data are mean ±SE
(bars) values of four measurements from the stria-
tum and two values from the substantia nigra from
each rat (n = 7 rats). °p< 0.05. Also shown are
representative sections of (B) preproenkephalin
mRNA, (D) preprotachykinin mRNA, and (F) tyrosine
hydroxylase mRNA in the caudate putamen (B and
D) and substantia nigra (F) of a control animal. White
represents high levels of binding.

We have previously shown by in vivo microdialysis
that Ang II stimulates dopamine release in the rat stria-
turn via a direct action on the dopaminergic terminals
(Mendelsohn et al., 1993) and induces turning in the
unilateral 6-hydroxydopamine-lesioned substantia ni-
gra model (Jenkins et al., 1995). In view of our previ-
ous findings, after chronic ACE inhibition, we ex-
pected Ang II levels to be decreased, thereby resulting
in a decline in dopamine levels. Our current results
TABLE 2. Effect of chronic ACE inhibition on tissue
contents of dopamine and metabolites
Content (~sg/gof tissue)
with chronic ACE inhibitor-treated rats displayed the
opposite effect to what was predicted, detecting an
increase in dopamine content and release.
We have morphological evidence that Ang II AT
1
receptors and ACE in the striatum are associated with
different neuronal pathways. Our autoradiographic
studies in humans (Allen et al., 1991, 1992) revealed
that Ang II receptors in the striatum are present on the
terminals of the nigrostriatal dopaminergic projections.
This distribution is in contrast to the ACE, which oc-
curs in striatopallidal and striatonigral neurons (Chai
et al., 1987a,b). I© addition to Ang II and bradykinin,
ACE is also able to cleave other small peptides in vitro,
including [Met I and [Leu] -en.kephalin, substance P,
-

and dynorphin, which occur in abundance in the stria-

ACE topallidal and striatonigral pathways.
Control inhibitor-treated p value
We have shown an increase in striatal preproenkeph-
Striatum alin mRNA level after chronic ACE inhibitor treat-
Dopamine 3.53 ±0.44 9.23 ±2.35 p < 0.05 ment. Centrally, ACE can contribute to the synthesis
DOPAC 2.14 ±0.21 5.23 ±1.85 p = 0.13 and metabolism of enkephalins (Yang et al., 1981;
HVA 2.56 ± 1.07 4.08 ±1.58 p = 0.45 Palenker et al., 1984; Lantz and Terenius, 1985; Kase
Frontal cortex
Dopamine 0.23 ±0.05 0.49 ±0.01 p = 0.09 et al., 1986). Interactions between ACE and the central
DOPAC 0.09 ±0.03 0.16 ±0.05 p = 0.26 enkephalinergic system have been observed. Intracere-
HVA 0.13 ±0.02 0.18 ±0.04 p = 0.25 broventricular administration of ACE inhibitors po-
tentiated the analgesic effects of intracerebroventricu-
Tissue contents of dopamine, DOPAC, and HVA were measured lar [Met]-enkephalin (Stine et al. 1980) (an effect
by HPLC before and after I week of ACE inhibition and compared
with values in untreated controls. Data are mean ±SEM values (n blocked by the opioid antagonist naltrexone) and 6- of
= 6). the more
Phe7 potent et
(Norman heptapeptide
al., 1985). [Met] -enkephalin-Arg
Clinically, a case study

J. Neurochem., Vol. 68, No. 3, 1997

 ACE INHIBITORS AND DOPAMINE
reported that psychosis induced by captopril adminis-
tration was immediately relieved by naloxone treat-
ment (Gillman and Sandyk, 1985), suggesting that the
inhibition of breakdown of enkephalins may have been
involved.
When infused into the substantia nigra, enkephalins
have been shown to increase striatal dopamine turnover
(Kalivas and Stewart, 1991). Moreover, disruption of
the nigrostriatal dopaminergic neurons increases the
levels of the dopamine D
2 receptor and [Met] -enkeph-
alin mRNAs that are colocalized in the striatopallidal
pathway. These observations suggest that these two
pathways may be intrinsically linked to modulate each
other’s activity. It thus appears that the ACE inhibitor
effect on dopamine release may be mediated, at least
in part, by increasing the level of preproenkephalin
mRNA within the striatum.
We saw no difference in striatal preprotachykinin
mRNA content between control and perindopril-
treated groups but cannot rule out the influence of
substance P in this study. After all, human ACE has
been reported to cleave substance P in vitro (Skidgel
et al., 1984), and an in vivo effect of ACE inhibitors
on the potentiation of substance P-induced salivation
in the rat has been reported (Cascieri et al., 1984).
Neuroanatomical studies reveal colocalization of dopa-
mine D1 receptor and substance P mRNAs in striatoni-
gral neurons (Gerfen, 1995). This suggests that ACE
inhibition may have less influence over the striatonigral
pathway than the indirect, striatopallidal pathway.
We cannot be certain, however, that these two pep-
tides alone are influencing the dopaminergic system.
ACE is capable of hydrolyzing other neuropeptides in
vitro that are known to interact with the nigrostriatal
dopamine system. These include neurotensin (Ruggeri
et al., 1987; Skidgel et al., 1987; Blaha et al., 1990)
and dynorphin (Benuck et al., 1984; Li et al., 1986,
1988).
We observed no changes in tyrosine hydroxylase
mRNA content after ACE inhibition, suggesting that
ACE inhibitors are not modulating dopamine content
via increasing tyrosine hydroxylase synthesis. The lack
of effect on the dopamine metabolites is difficult to
interpret. In our previous microdialysis studies we have
shown that Ang II stimulates the release of dopamine,
DOPAC, and HVA from the rat striatum (Mendelsohn
et a!., 1993). However, the dopamine metabolites may
not always move parallel to dopamine.
The possibility that the chronic effect of ACE inhibi-
tors on reducing the extent of gliosis around the micro-
dialysis probe and thereby resulting in an increase in
the detection of dopamine is not supported by our ob-
servation that there is no difference in GFAP staining
in the area of the striatum surrounding the implanted
microdialysis probe between the treated and control
groups. Further support for a specific effect of ACE
inhibitors on increasing dopamine release is the data
from our complementary study on striatal extract
1309
where an increase in dopamine content was seen in
the intact brain after chronic ACE inhibitor treatment.
Clinically, several reports have suggested that ACE
inhibitors induce a feeling of well-being and improve
quality of life in hypertensive patients (Beto and Ban-
sal, 1992). In a study of three depressed patients, cap-
topril treatment was associated with substantial mood
elevation (Zubenko and Nixon, 1984). The effect of
ACE inhibitors on cognition is uncertain. In healthy
male subjects, single doses of captopril have been re-
ported to improve short-term memory and perfor-
mances at letter cancellation tests (Currie et al., 1990).
On the other hand, other authors found no effects of
ACE inhibitors on cognition (Bulpitt and Fletcher,
1992).
The involvement of ACE inhibitors in the modula-
tion of central neurotransmitters has not been well in-
vestigated, although these drugs affect cognitive func-
tion and mood. Moreover, the possibility that ACE
inhibitors can regulate the levels of a range of small
neuropeptides other than Ang II and bradykinin sug-
gests a more important role for ACE in the brain than
initially suspected.
The findings of this study confirm that chronic ACE
inhibitor treatment does indeed influence the levels of
dopamine in the brain. The full potential of these drugs
as therapeutic agents in the treatment of syndromes
involving the brain dopaminergic system needs to be
investigated further.
Acknowledgment: This work is funded by the National
Health and Medical Research Council of Australia, National
Heart Foundation, and Austin Hospital Medical Research
Foundation. T.A.J. holds an NH&MRC Dora Lush Postgrad-
uate Biomedical Scholarship. The authors would like to
thank Ms. Carmel Murone and Mr. Gabriel Liberatore for
excellent technical assistance. The ACE inhibitor 35 1A was
a kind gift from Dr. C. S. Sweet, Merck Institute, Rahway,
NJ, U.S.A.
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