Relationship between ace genotype and short duration aerobic performance development

Abstract

We have previously demonstrated that, ACE D allele may be related with a better performance in short duration aerobic endurance in a homogeneous cohort with similar training backgrounds. We aimed to study the variation in the short-duration aerobic performance development amongst ACE genotypes in response to identical training programs in homogeneous populations. The study group consisted of 186 male Caucasian non-elite Turkish army recruits. All subjects had undergone an identical training program with double training session per day and 6 days a week for 6 months. Performances for middle distance runs (2,400 m) were evaluated on an athletics track before and after the training period. ACE gene polymorphisms were studied by PCR analysis. The distribution of genotypes in the whole group was 16.7% II, n=31; 46.2% ID, n=86; 37.1% DD, n=69. Subjects with ACE DD genotype had significantly higher enhancement than the ID (P<0.01) and II (P<0.05) genotype groups. Around 2,400 m performance enhancement ratios showed a linear trend as ACE DD>ACE ID>ACE II (P value for Pearson chi2=0.461 and P value for linear by linear association=0.001). ACE DD genotype seems to have an advantage in development in short-duration aerobic performance. This data in unison with the data that we have obtained from homogenous cohorts previously is considered as an existence of threshold for initiation of ACE I allele effectiveness in endurance performance. This threshold may be anywhere between 10 and 30 min with lasting maximal exercises.

Full text

Eur J Appl Physiol
DOI 10.1007/s00421-006-0286-6
123
ORIGINAL ARTICLE
Relationship between ace genotype and short duration aerobic
performance development
Mesut Cerit · MuzaVer Colakoglu · Murat Erdogan ·
AWg Berdeli · Fethi Sirri Cam
Accepted: 27 July 2006
© Springer-Verlag 2006
Abstract We have previously demonstrated that,
ACE D allele may be related with a better perfor-
mance in short duration aerobic endurance in a homo-
geneous cohort with similar training backgrounds. We
aimed to study the variation in the short-duration aero-
bic performance development amongst ACE geno-
types in response to identical training programs in
homogeneous populations. The study group consisted
of 186 male Caucasian non-elite Turkish army recruits.
All subjects had undergone an identical training pro-
gram with double training session per day and 6 days a
week for 6 months. Performances for middle distance
runs (2,400 m) were evaluated on an athletics track
before and after the training period. ACE gene poly-
morphisms were studied by PCR analysis. The distri-
bution of genotypes in the whole group was 16.7% II,
n = 31; 46.2% ID, n =86; 37.1% DD, n = 69. Subjects
with ACE DD genotype had signiWcantly higher
enhancement than the ID (P < 0.01) and II (P <0.05)
genotype groups. Around 2,400 m performance
enhancement ratios showed a linear trend as ACE
DD > ACE ID > ACE II (P value for Pearson

2
= 0.461 and P value for linear by linear
association = 0.001). ACE DD genotype seems to have
an advantage in development in short-duration aerobic
performance. This data in unison with the data that we
have obtained from homogenous cohorts previously is
considered as an existence of threshold for initiation of
ACE I allele eVectiveness in endurance performance.
This threshold may be anywhere between 10 and
30 min with lasting maximal exercises.
Keywords Genetics · Endurance · Insertion/deletion ·
Training · ACE
Introduction
High performance in short duration aerobic perfor-
mance (2–8 min) demands high power output and
increased tissue oxygenation. It requires higher VO
2max
and strength endurance levels. Angiotensin I-convert-
ing enzyme (ACE) cleaves vasodilator kinins while
promoting formation of the vasoconstrictor angioten-
sin II. Increased plasma angiotensin II levels restrict
blood Xow to tissues. The human ACE gene contains a
polymorphism consisting of the presence (insertion, I)
or absence (deletion, D) of a 287 base pair sequence in
M. Cerit
Institute of Health Sciences, Sport Sciences Division,
Ege University, Izmir, Turkey
M. Colakoglu (&)
School of Physical Education and Health,
Department of Coaching Education, Ege University,
Beden Egitimi ve Spor Yuksekokulu,
35100 Bornova, Izmir, Turkey
e-mail: colakoglu@besyo.ege.edu.tr
M. Erdogan
Institute of Health Sciences,
Physical Education and Sports Division,
Gazi University, Ankara, Turkey
A. Berdeli
Faculty of Medicine, Department of Pediatrics,
Molecular Diagnostics Laboratory, Ege University,
Izmir, Turkey
F. S. Cam
Faculty of Medicine,
Department of Medical Biology and Genetics,
Celal Bayar University, Manisa, Turkey

Eur J Appl Physiol
123
intron 16 (Rigat et al. 1990). This polymorphism seems
to have an important role on ACE at a cellular level
(Davis et al. 2000; Mizuiri et al. 1997) and may eVect
angiotensin II production.
The present data on the ACE I/D polymorphism
and exercise performance are somewhat controversial.
The ACE I-allele usually seems to be associated with
enhanced aerobic endurance performance (Alvarez
et al. 2000; Gayagay et al. 1998; Montgomery et al.
1998; Myerson et al. 1999; Nazarov et al. 2001). How-
ever, in some studies higher VO
2max
levels, which indi-
cate an improved oxidative capacity, found to be
related with ACE D-allele (Rankinen et al. 2000a;
Zhao et al. 2003). On the other hand, ACE D-allele is
related with higher fast-twitch (FT) muscle Wber ratio
(Zhang et al. 2003), greater strength gain in the quadri-
ceps muscle in response to training (Folland et al.
2000), and better anaerobic performance (Woods et al.
2001). In contrast, some researchers have not found a
relationship between ACE genotype and athletic per-
formance in elite athletes (Rankinen et al. 2000b; Tay-
lor et al. 1999), and sedentary subjects (Rankinen et al.
2000a).
Such associations with athletic performance and
ACE I/D polymorphism have been replicated across
diVerent races, geographical locations, athletic status
and sporting disciplines (Alvarez et al. 2000; Myerson
et al. 1999; Woods et al. 2001). Studies of those of
mixed ability and mixed sporting disciplines have thus
tended to be negative (Woods et al. 2001) as have
those confounded by a mixture of those of diVerent
race and sex or training regimen (Nazarov et al. 2001;
Taylor et al. 1999).
We have previously demonstrated that ACE D
allele may be related with a better performance in
short duration aerobic endurance in a homogeneous
cohort (Cam et al. 2005). However, the study was
cross-sectional and the group was small (n =88).
We postulated that ACE D allele is associated with
a better short-duration aerobic performance develop-
ment in response to identical training programs in
homogeneous populations. To clarify this hypothesis,
we aimed to study the variation in the performance as a
result of 6 months endurance training in the army
recruits.
Methods
Subjects
The study group consisted of 186 male Caucasian non-
elite Turkish army recruits. The study had appropriate
ethics committee approval. Written informed consent
was obtained from all participants.
Training program
All subjects had undergone an identical training pro-
gram with double training session per day and 6 days a
week for 6 months. The program consists of Xexibility
exercises, circuit trainings, 2,400 and/or 3,000 m runs,
1,000–3,000 m runs with military equipment, hurdling
course, aerobic threshold and anaerobic threshold tra-
inings. The circuit trainings were consisted of gallows,
sit-ups, push-ups and rope-climbs, bomb throws, hur-
dling course. In initial 2 weeks, there were approxi-
mately 30 min whole body Xexibility exercises and
circuit trainings every weekday, 30–45 min anaerobic
threshold runs and 45–60 min aerobic threshold runs
alternately except Sundays. From third week onwards,
one hurdling course training, and one or two of the
1,000 –3,000 m run with military equipment and/or the
2,400 or 3,000 m running were replaced with one of the
aerobic or anaerobic threshold training.
Exercise tests
Performances for middle distance runs (2,400 m) were
evaluated on an athletics track before and after the
training period. Performance times were determined
with digital timers in 0.01 s accuracy by three referees.
The time in the middle was recorded.
Genetic analysis
Genomic DNA was extracted from 200 l of EDTA-
anticoagulated peripheral blood leucocytes using the
QIAmp Blood Kit (QIAGEN, Ontario, Canada, Cat.
no:51,106). AmpliWcation of DNA for genotyping the
ACE I/D polymorphism was carried out by polymerase
chain reaction (PCR) in a Wnal volume of 15 l contain-
ing 200 M dNTP mix, 1.5 mM MgCl
2
, 1£ BuVer, 1 unit
of AmpliTaq
®
polymerase (PE Applied Biosystems)
and 10 pmol of each primer. The primers used to encom-
pass the polymorphic region of the ACE were 5⬘-CTGG
AGACCACTCCCATCCTTTCT-3⬘ and 5⬘-ATGTGG
CCATCACATTCGTCAGAT-3⬘ (Rigat et al. 1992).
DNA is ampliWed for 35 cycles, each cycle comprising
denaturation at 94°C for 30 s, annealing at 50°C for 30 s,
extension at 72°C for 1 min with Wnal extension time of
7 min. The initial denaturizing stage was carried out at
95°C for 5 min. The PCR products were separated on
2.5% agarose gel and identiWed by ethidium-bromide
staining. Each DD genotype was conWrmed through a
second PCR with primers speciWc for the insertion

Eur J Appl Physiol
123
sequence (Shanmugam et al. 1993). The samples with II
and DD homozygote genotypes and ID heterozygote
genotype were selected at random. These samples were
then puriWed by PCR products puriWed system (Genomics,
Montage PCR, Millipore) and directly sequenced by the
ABI 310 Genetic Analyzer (ABI Prisma PE Applied
Biosystems).
Statistical analysis
Statistical analyses were performed using SPSS for Win-
dows version 12.0 (SPSS Inc., Chicago, IL, USA). Meth-
ods applied were frequencies, cross-tabulations,
descriptive statistics, and means. Statistical signiWcance
was set at the P < 0.05 level. A 
2
test with the data read
from Finetti statistics program was used to conWrm that
the observed genotype frequencies were in Hardy–
Weinberg equilibrium. DiVerences amongst ACE geno-
type groups in endurance performance were tested with
analysis of variance (ANOVA) and post-hoc Bonferroni
test. Genotype distribution across performance levels
was compared by chi-square for linear trend. DiVerences
between baseline and post-training values of each ACE
genotype group were analyzed by t-test.
Results
The distribution of genotypes in the whole group
(16.7% II, n = 31; 46.2% ID, n = 86; 37.1% DD, n =69)
did not deviate signiWcantly from those predicted by
the Hardy–Weinberg equilibrium. The allele frequen-
cies of the subjects were 0.398 and 0.602 for the I and D
alleles, respectively. Baseline 2,400 m performance lev-
els were not diVerent amongst ACE genotype groups
(Table 1).
All ACE genotype groups showed signiWcant
improvements in 2,400 m performance after training
period as compared to baseline levels (P <0.001 for all).
However, subjects with ACE DD genotype had signiW-
cantly higher enhancement than the ID (P <0.01) and II
(P < 0.05) genotype groups (Table 1, 2). Around
2,400 m performance enhancement ratios (variation %)
showed a linear trend as ACE DD > ACE ID > ACE II
(P value for Pearson 
2
= 0.461 and P value for linear by
linear association = 0.001).
Discussion
We have previously reported that ACE D allele may
be related with a better performance in short-duration
aerobic endurance (2,000 m) in a homogeneous cohort
(Cam et al. 2005) and, also found that I allele responses
better to medium-duration (30 min) aerobic endurance
training (Cam et al. 2006).
In this study, we demonstrated that ACE DD geno-
type has an advantage in short-duration aerobic endur-
ance (2,400 m) development in response to training.
Thus, it seems that the initiation of the eVectiveness of
ACE I allele in better performances or responses to
training in endurance events is somewhere between
approximately 10 –30 min.
High level of power production, VO
2max
and anaero-
bic capacity is necessary for success in middle distance
running performances. VO
2max
levels can be sustained
10–12 min (Martin 1990). Since our subjects baseline
performances are close to 10 min and post-training
performances are better, it suggests that their exertion
is at least equal or even higher than VO
2max
. Running
performances corresponding to VO
2max
resulted in 8–
12 mM blood lactate concentrations (Noakes 1988).
Ohkuwa et al. (1984) had shown that mean peak blood
lactate levels were 12 mM after an exhaustive 3,000 m
running in track and Weld athletes. Thus, it may be pos-
tulated a high anaerobic energy contribution exists in
2,400 m maximal running performance.
Table 1 DiVerences amongst
A
CE genotype groups in
2,400 m performance
ACE II ACE ID ACE DD ANOVA
(n = 31) (n =86) (n =69) 
2
P
Baseline (s) 599.1 § 33.0 591.9 § 33.9 601.0 § 40.3 1.305 0.274
Post-training (s) 541.0 § 25.4 532.5 § 28.0 529.6 § 28.7 1.809 0.167
P (t-test) 0.0001** 0.0001** 0.0001**
Variation
a
(%) 9.59 § 3.64 9.94 § 3.7 11.6 § 3.4 6.669 0.02*
*P < 0.05; **P <0.01
a
{1-[last value/previous
value]} £100
Table
2
D
iV
erences
i
n 2,400 m per
f
ormance
i
mprovements
amongst ACE genotype groups
*P < 0.05; **P < 0.01(Bonferroni test)
ACE II,
(n =31)
ACE ID,
(n =86)
ACE DD,
(n =69)
A
CE II – 1.000 0.013*
A
CE ID 1.000 – 0.004**

Eur J Appl Physiol
123
ACE D allele seems related with a higher VO
2max
(Rankinen et al. 2000a; Zhao et al. 2003) and superior
performances in middle and long distance swimming
(Tsianos 2004). ACE DD genotype may be associated
with a greater skeletal muscle strength gain in response
to training (Colakoglu et al. 2005; Folland et al. 2000;
Hopkinson et al. 2004) and a higher anaerobic capacity
(Woods et al. 2001). This genotype is found to be related
to a higher percentage of type-II muscle Wbers (Zhang
et al. 2003). Middle distance runners (800–3,000 m) have
a relatively high percentage (48–55%) of fast-twitch
Wbers (Noakes 1991). Therefore, ACE DD genotype
subjects may have an advantage in short-duration aero-
bic performances that requires high level VO
2max
.
Indeed, recent data have some conXictions on the
eVectiveness of ACE I/D polymorphism and exercise
performance. Besides many research projects revealing
that there may be an association between ACE I/D
polymorphism and athletic performance, Rankinen
et al. (2000b) concluded that there was no relationship
between ACE I/D polymorphism and elite athlete sta-
tus in 192 athletes whose VO
2max
was at least
75 ml kg
¡1
min
¡1
. Likewise, Taylor et al. (1999) did not
Wnd any association between ACE I/D polymorphism
and athletic performance in a cohort, composed with
both genders. However, they found a trend toward the
DD genotype in males but the trend was inconsistent in
females. Also, Sonna et al. (2001) have reported that
ACE genotype was not strongly related to physical per-
formance in their studies on the eVect of training on
aerobic power and muscular endurance in 147 healthy
US Army recruits of diVerent ethnicity.
Conclusion
ACE DD genotype seems to have an advantage in
development in shortduration aerobic performance.
There was also a linear trend in performance enhance-
ment as ACE DD > ID > II. This data in unison with
the data that we have obtained from homogenous
cohorts previously is considered as an existence of
threshold for initiation of ACE I allele eVectiveness in
endurance performance. This threshold may be any-
where between 10 and 30 min lasting maximal exer-
cises.
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