Osteoporos Int (2007) 18:1235–1242
DOI 10.1007/s00198-007-0374-4
ORIGINAL ARTICLE
Polymorphisms in the vitamin D receptor gene are associated
with muscle strength in men and women
A. Windelinckx & G. De Mars & G. Beunen & J. Aerssens &
C. Delecluse & J. Lefevre & M. A. I. Thomis
Received: 16 December 2006 / Accepted: 16 March 2007 / Published online: 4 April 2007
# International Osteoporosis Foundation and National Osteoporosis Foundation 2007
Abstract
Introduction Vitamin D receptor (VDR) polymorphisms
have been associated with fracture risk and muscle strength,
although evidence for the latter is limited and conflicting.
Methods BsmI, TaqI and FokI VDR polymorphisms were
genotyped in 253 men (54.9±10.2 yr) and 240 women (41.5±
13.2 yr). Haplotypes were constructed for BsmI and TaqI.
Handgrip, isometric (at 60°, 120° and 180° joint angle) and
eccentric torques (60°/s) of knee extension and flexion were
analysed using AN(C)OVA. Torque-velocity curves were
constructed for concentric torques at 60°/s, 180°/s and 240°/s
and analysed using multivariate AN(C)OVA. Age, height
and fat-free mass were included as covariates.
VDR is a candidate gene for interindividual muscle strength
variability. Here we studied three common polymorphisms in relation
to isometric and dynamic knee strength in 253 men and 240 women.
An association was found for quadriceps strength with FokI in women
and with BsmI-TaqI haplotype in men.
A. Windelinckx : G. De Mars : G. Beunen : C. Delecluse :
J. Lefevre : M. A. I. Thomis
Research Center for Exercise and Health,
Department of Biomedical Kinesiology,
Faculty of Kinesiology and Rehabilitation Sciences,
Katholieke Universiteit Leuven,
B-3001 Leuven, Belgium
M. A. I. Thomis
e-mail: Martine.Thomis@faber.kuleuven.be
J. Aerssens
Department of Translational Medical Research,
TIBOTEC bvba,
B-2800 Mechelen, Belgium
A. Windelinckx (*)
Tervuursevest 101,
B-3001 Heverlee, Belgium
e-mail: An.Windelinckx@faber.kuleuven.be
Results Quadriceps isometric and concentric strength were
higher in female f/f homozygotes compared to F allele
carriers. Adjustment for confounding factors rendered results
for quadriceps isometric strength at 120° non-significant. No
significant association was found with BsmI-TaqI haplotype
in women. In contrast, male Bt/Bt homozygotes had higher
isometric quadriceps strength at 150° and higher concentric
quadriceps strength than bT allele carriers without and with
adjustment for confounding factors. No association was
observed with FokI in men. In both genders, no interaction
effect was present between BsmI-TaqI haplotype and FokI.
Conclusions Different VDR gene polymorphisms are asso-
ciated with quadriceps strength in men and women.
Keywords Ageing . Association analysis .
Gender specific effects . Muscle strength . VDR
Introduction
Fragility-caused bone fractures are a major health care
issue. Attempts have been made to determine risk factors
that predict an individual’s bone fracture risk. Compro-
mised bone strength, resulting from a low bone mass and
density (osteoporosis), as well as falling, alone or in
combination, have been recognized as two independent
and immediate risk factors of fracture in elderly. In
addition, several other, less pronounced risk factors have
been identified, such as aging, inactivity, poor nutrition,
smoking, genetic predisposition, use of alcohol, diseases,
medications, functional impairments, and disabilities [1].
Vitamin D is well known for its regulatory role in calcium
and phosphate homeostasis and can mediate its effect through
the formation of a complex with the vitamin D receptor
(VDR). As both calcium and phosphate are involved in bone
1236
metabolism, most of the initial genetic studies focussed on the
association between VDR gene polymorphisms and bone
mineral density (BMD). Results are, however, conflicting:
some investigators found an association [2–7], while others
did not [8–12]. More recently, associations between VDR
polymorphisms and fracture risk were reported independent-
ly from BMD [13–16].
In contrast to the abundance of studies on VDR poly-
morphisms and BMD, only a few studies have investigated
the relationship between VDR genotypes and the second
major fracture risk factor, namely falling. Falling can be
related to two major underlying factors: impaired balance
and muscle weakness [16–21]. In the present study, we
focus on the relation between VDR genotypes and muscle
strength. It has previously been shown that VDR is
expressed in muscle [22] and polymorphisms within the
VDR gene can affect strength. Geusens et al. [23, 24]
reported a significant association between the BsmI geno-
type and quadriceps and grip strength in elderly non-obese
women (25% and 7% lower in BB compared with bb
genotype, respectively). Neither Dukas et al. [25] nor
Grundberg et al. [26] could replicate these findings. They
did, however, find a correlation of the BsmI genotype with
hamstrings strength in elderly and adult women, with BB
individuals having the highest strength, followed by Bb and
bb. Additionaly, Dukas et al. [25] found the same pattern of
association for handgrip strength. Young Chinese female B
allele carriers showed significantly higher peak torque in
concentric knee flexors than bb homozygotes and ApaI null
allele carriers (AA) exhibited significantly lower knee and
elbow concentric or eccentric peak torque than the a allele
carriers [27]. Until now, no association has been found
between the FokI, BsmI, ApaI or TaqI polymorphisms and
muscle strength in elderly male populations [5, 25, 28],
although Van Pottelbergh et al. [5] reported a solitary
association between FokI genotype and lower limb strength
(measured by standing time) in a sample of adult men.
Since studies on the relationship between VDR geno-
types and muscle strength are limited, especially for the
TaqI and FokI polymorphims, and results from association
studies with BsmI are ambiguous, we performed an
association analysis on the BsmI, TaqI and FokI VDR
polymorphisms with muscle strength (handgrip and iso-
metric and dynamic knee strength) in a cohort of 253 men
and 240 women.
Material and methods
Subjects
Subjects were healthy volunteers from a study conducted
within the framework of the Policy Research Centre Sport,
Osteoporos Int (2007) 18:1235–1242
Physical Activity and Health. Cohort 1 comprises 154 men
(age 45–49 yr) and 138 women (age 38–44 yr) as a follow-
up of the Leuven Longitudinal Study on Lifestyle, Physical
Fitness and Health. Cohort 2 was recruited via socio-
cultural organisations and via advertisements in local and
national media in the region of Leuven (Belgium) as part of
a comparative physical fitness intervention study. Subjects
were at least 60 years old, had not participated in any
endurance and/or strength training in a period of 2 years
preceding the study, and did less than 2 hours/week of
moderate physical activity at the moment of recruitment.
Subjects with pathologies contra-indicative to participation
in a training program, with neurodegenerative or neuro-
muscular diseases or taking drugs that influence exercise
capacity or bone density were excluded. Pre-training
evaluations were used in this study. Both studies were
approved by the medical and ethical committee of the
Katholieke Universiteit Leuven and all subjects gave
written informed consent before participating in the study.
Participants of both cohorts were measured with exactly the
same methods regarding anthropometric and strength
characteristics (see below).
Body composition
A comprehensive set of skinfolds and circumferences was
taken by experienced anthropometrists as described in detail
elsewhere [29]. Fat-free mass (FFM) was estimated by the
Durnin and Womersley equation [30], for which body weight
and skinfolds of triceps, biceps, suprailiac, and subscapula
were used. Body mass index (BMI) was calculated as an
estimate of obesity (weight [kg] / (height [m])2).
Strength measurements
Hand grip strength was measured using a Jamar handgrip
dynamometer (Sammons Preston). The strength of the
knee flexors and extensors was evaluated using an
isokinetic dynamometer (Biodex, Biodex Medical Sys-
tems, Inc.). All tests were performed unilaterally on the
right side, unless the right knee had once been injured.
Subjects were seated on a backwards inclined seat (15°)
with the upper legs, hips and shoulders stabilized using
safety belts. The rotation axis of the joint was aligned
with the mechanical axis of the dynamometer and full
extension of the knee is considered as 180°. Subjects were
verbally encouraged to perform at their maximum effort
and visual feedback of their performance was presented
after each test.
Maximal isometric knee extension strength was mea-
sured at 150°, 120° and 90°. The subjects performed two
isometric contractions at each angle for five seconds,
separated by a 10 s rest period. Between trials for each
Osteoporos Int (2007) 18:1235–1242
angle there was a rest period of 20 s. The highest torque
(Nm) of the two trials was recorded as the maximal
isometric strength. Maximal isometric knee flexion strength
was measured at 120°.
Subjects performed two series of consecutive isokinetic
extension-flexion movements against the lever arm with a
determined velocity of 60°/s (4 repetitions) and 240°/s (six
repetitions) between a knee angle of 90° and 160°. Between
the two series there was a rest period of 20 seconds. The
maximal dynamic extension and flexion strength was
determined as the peak torque (Nm) during these series of
knee extension and flexion.
Maximal eccentric knee extension and flexion strength
was tested at 60°/s with two series of consecutive flexion-
extension movements against the lever arm. The maximal
eccentric flexion and extension torque (Nm) was retained
for further analyses.
Finally, subjects performed a series of 25 consecutive
extension and flexion movements between a knee-angle of
90° and 160° with a determined velocity of 180°/s. Again,
maximal dynamic extension and flexion strength was
determined as the peak torque (Nm).
Genotyping
Genomic DNA was extracted from ethylenediamine-tetra-
acetic acid (EDTA) whole blood using a salting-out
procedure. Genotyping of the following VDR gene poly-
morphisms was performed using a Sequenom MassARRAY
System (Sequenom, Inc.): FokI (rs2228570) in exon 1,
BsmI (rs2544410) in intron 8 and TaqI (rs731236) a syn-
onymous SNP in exon 9 of the gene [31]. These poly-
morphisms were named after the restriction enzyme that
have traditionally been used for their detection; the
presence of a restriction site is conventionally denoted with
a lower case letter (f, b and t for FokI, BsmI and TaqI,
respectively), whereas upper case letters (F, B and T)
indicate the absence of the restriction site.
Haplotypes for the BsmI and TaqI polymorphisms were
inferred using PHASE version 2.1.1 software, freely avail-
able at http://www.stat.washington.edu/stephens/software.
html. For ambiguous pairs of haplotypes, the most likely
pair was selected.
Statistical methods
All statistical analyses were performed separately for men
and women. Chi-square tests were used to assess deviations
from Hardy-Weinberg equilibrium. Analyses of (co)vari-
ance (AN(C)OVA) were used to compare muscle strength
measurements between genotype groups. To reduce the
number of tests, dynamic torque-velocity curves were
constructed from the concentric measurements at 60°/s,
1237
180°/s and 240°/s. When the velocities are logarithmically
transformed these curves become linear and can therefore
be analysed using a general linear models approach.
Eccentric and isometric torques can not be included in this
linear relationship and are analysed separately using an AN
(C)OVA approach. All analyses are performed without and
with adjustment for potential confounding factors (age,
height and total fat-free mass). Interaction effects between
the BsmI-TaqI haplotype and the FokI polymorphism were
tested using pre-planned contrasts. Results are expressed as
(least square) means ± standard error (SEM) and shown in
Tables and Figures with adjustment for age, height and total
fat-free mass unless stated otherwise. The level of signif-
icance was set at p < 0.05. Power calculations were
performed based on estimated effect sizes and observed
sample sizes using proc power. For the present sample
sizes, an effect size of 0.82 can be detected with 80%
power. This corresponds to 10–12% of overall explained
variance and therefore to a substantial genetic effect. All
statistical analyses were performed in SAS version 9.1
(SAS Institute Inc.).
Results
Distribution of VDR genotypes
The genotype distributions of the BsmI and TaqI geno-
types were in Hardy-Weinberg equilibrium (HWE) for
men and women whereas the FokI genotype distribution
was in HWE only for men, not for women (Table 1). As
previously described by Morisson et al. [3], TaqI and BsmI
VDR polymorphisms are linked (chi-square test for
independence; df=4; all p<0.0001; see Table 2). There-
fore haplotypes were constructed for further analyses; four
different haplotype alleles resulting in seven different
haplotype genotypes were found (Table 1). Only the most
common haplotype alleles (i.e., with allele frequency
> 5%; haplotype alleles bT and Bt) and their resulting
genotypes were retained for further analyses. As results
from analyses of both polymorphisms separately are
comparable to those from haplotype analyses, only the
latter are shown.
Descriptive statistics
Subject characteristics for men and women are provided in
Table 3. No significant differences were observed between
any of the VDR genotypes and any of the descriptive
statistics in men. In women, however, there was a
statistically significant association between FokI genotype
and height: f/f homozygotes were taller than F allele
carriers (165.7±1.0 vs. 163.2±0.5; p=0.03).
1238 Osteoporos Int (2007) 18:1235–1242

Table 1 Allele, haplotype and genotype distributions of TaqI, BsmI analysing allelic association with f/f homozygotes being
and FokI VDR gene polymorphisms
stronger than F allele carriers (p<0.05 for both 90° and 120°).
Men (253) Women (240) Knee extensor and flexor torque-velocity curves are
shown in Fig. 1. No significant association with the FokI
Alleles polymorphism was found for knee flexors and for eccentric
TaqIa
knee extensor strength. For the concentric torque-velocity
T (T) 306 (60.7)b 271 (56.9)c
t (C) 198 (39.3)b 205 (43.1)c curves of the knee extensors multivariate ANOVA indicates
BsmIa a trend towards an overall genotype effect (intercept effect)
B (A) 200 (39.7)b 210 (43.8) with f/F heterozygotes having less strength than both
b (G) 304 (60.3)b 270 (56.3) homozygote groups (p=0.1). However, no significant
FokIa genotype*velocity interactions (slope effect) were present.
F (C) 314 (62.3)b 309 (64.6)b When analysing allelic association, F allele carriers had
f (T) 190 (37.7)b 169 (35.4)b
marginally less concentric quadriceps strength than f/f
Haplotype allelesa
homozygotes (p=0.06).
bT (GT) 302 (59.7) 269 (56.1)
Bt (AC) 196 (38.7) 207 (43.1) To investigate whether the VDR genotype effects on
BT (AT) 6 (1.2) 3 (0.6) muscle strength were modified by confounding factors, age,
bt (GC) 2 (0.4) 1 (0.2) height and total fat-free mass were included as covariates in
Haplotype genotypesa an ANCOVA analysis. Taking these confounding factors
Bt-bT (AC/GT) 124 (49.0) 117 (48.8) into account, we find that the results demonstrated
bT-bT (GT/GT) 87 (34.4) (30.8) 74 marginally significant genotypic associations and signifi-
Bt-Bt (AC/AC) 35 (13.8) (18.8) 45
cant differences between f/f homozygotes and F allele
BT-bT (AT/GT) 2 (0.8) 3 (1.2)
bt-bT (GC/GT) 2 (0.8) 1 (0.4)
carriers for isometric knee extensor strength at 90°. The
Bt-BT (AC/AT) 2 (0.8) 0 (0.0) difference in isometric strength at 120° becomes non-
BT-BT (AT/AT) 1 (0.4) 0 (0.0) significant. For dynamic knee extensor torques a trend
towards an overall genotypic effect is present in both
Values are counts with percentages between brackets; genotypic and allelic analyses (p<0.1).
a: Common notation with corresponding nucleotide between brackets;
b: 1 missing genotype; c: 2 missing genotypes
Men
VDR genotype and muscle strength
A borderline non-significant trend (p=0.06) was observed
Women for handgrip with Bt/Bt homozygotes having higher
strength than bT carriers (see Table 3 and Fig. 2). For the
No significant associations were found between the BsmI- isometric knee extensor strength an association was found
TaqI haplotype and any of the strength measurements (data for measurements at an angle of 150°, and a trend for 120°
not shown). Handgrip strength and isometric knee extensor (p=0.01 and p=0.06, respectively). When allelic association
and flexor strength in relation to the FokI polymorphism are was tested, Bt/Bt homozygotes had significantly higher
shown in Table 3. No significant differences between FokI isometric (at 150° and 120°) strength than bT carriers. No
genotype groups were present for handgrip, isometric knee significant differences were present for isometric flexor
extensor strength at 150° and isometric knee flexor strength strength. For the concentric knee extensor strength a non-
at all angles. However, knee extensor isometric strength at significant trend was observed when comparing the three
120° and 90° differed between genotype groups with the f/F genotype groups (p=0.1). When allelic association was
heterozygotes having less strength than both homozygotes. tested, Bt/Bt homozygotes had significantly higher concen-
This difference in isometric strength is also present when tric knee extensor strength than bT carriers. As in women,

Table 2 Chi-square test

for independence between Men Women
BsmI and TaqI genotypes
bb bB BB bb bB BB

tt 0 0 34 tt 0 0 44
tT 2 74 2 tT 1 116 0
TT 86 2 1 TT 74 3 0
χ2 359.76 p <0.0001 χ2 455.96 p <0.0001
df 4 df 4
Osteoporos Int (2007) 18:1235–1242 1239

Table 3 Subject characteristics, handgrip and isometric peak torque values for the knee extensors in women and men stratified according to
genotype

Women Men
c
f/f f/F F/F unadj. adj. p-value bT/bT bT/Bt Bt/Bt unadj. adj.
p-valueb p-valueb p-valuec

Subjects, n 39 91 109 87 124 35

Agea, yrs 51.8±12.9 51.8±13.5 51.1±12.9 0.93 55.4±10.4 55.0±10.0 52.7±9.96 0.40
Heigtha, cm 165.7±7.5 162.6±5.7 163.7±6.8 0.04d 175.7±6.3 176.4±6.2 175.9±5.9 0.86
Weigtha, kg 68.3±9.1 65.7±9.6 67.5±11.0 0.28 81.8±11.3 81.3±11.8 82.4±7.8 0.70
BMIa, kg/m2 24.9±3.0 24.9±4.0 25.3±4.5 0.76 26.5±3.5 26.1±3.6 26.7±2.4 0.60
Total FFMa, kg 44.7±4.4 44.2±4.4 44.7±5.0 0.70 62.1±6.1 62.2±5.9 62.7±4.4 0.90
Handgrip, kg 31.2±0.9 30.2±0.6 30.1±0.6 0.39 0.59 45.5±1.0 43.7±0.8 47.6±1.7 0.06 0.07
Knee Extensors
Isometric 90°, Nm 139.7±4.6 126.9±3.1 131.8±2.8 0.04d 0.07d 216.0±5.1 211.4±4.2 226.1±7.8 0.32 0.41
Isometric 120°, Nm 120.8±4.2 112.8±2.8 114.7±2.7 0.05d 0.29 174.0±4.2 170.4±3.4 187.6±6.3 0.06e 0.06e
Isometric 150°, Nm 62.7±2.6 60.1±1.7 61.2±1.6 0.32 0.70 88.8±2.1 83.8±1.8 95.5±3.3 0.02e 0.01e
Knee Flexors
Isometric 120°, Nm 70.3±2.2 68.2±1.5 69.5±1.4 0.15 0.68 116.7±2.6 115.8±2.1 123.3±4.0 0.24 0.33

a: Values are mean ± SD; BMI = body mass index; FFM = fat-free mass; b: p-value from the overall ANOVA comparing the three genotype
groups; c: p-value from ANCOVA adjusted for age, height and total fat-free mass comparing the three genotype groups; d: p<0.05 f/f vs.
F allele carriers; e: p<0.05 Bt/Bt vs. bT allele carriers

no significant interaction between genotype and velocity Discussion

was found. Neither for eccentric knee extensor strength nor
for any of the knee flexor strength measurements differ-
ences between genotype groups were observed. For the
FokI genotype, no significant associations were observed
for any of the strength measurements.
Adjustment of the analyses for age, height and total fat-
free mass did not alter results.
For both men and women no interaction effect between
the BsmI-TaqI haplotype and FokI was detected.
To our knowledge this is the first study to investigate VDR
polymorphisms in relation to both isometric and dynamic
knee strength. Our results show that both isometric and
concentric strength of the quadriceps can be influenced by
VDR genotypes, albeit by different polymorphisms in men
and women. For men, a trend towards a difference in
handgrip strength between BsmI-TaqI haplotype groups is
also present.

225 Bt/Bt
225 f/f * bT/bT
F/F 200
bT/Bt
200 f/F 175
175
Torque, Nm

150 #
Torque, Nm

150 ‡ 125 extensors

125
100
100
extensors
75
75
50 flexors
50
25 eccentric concentric
25 flexors
eccentric concentric 0
0 60 0 60 120 180 240
60 0 60 120 180 240
Velocity, ˚/s
Velocity, ˚/s Fig. 2 Torque-velocity curves in men, stratified according to BsmI/
Fig. 1 Torque-velocity curves in women, stratified according to FokI TaqI haplotype. 0°/s = isometric knee strength at 120°;* bT allele
genotype. 0°/s=isometric knee strength at 120°;‡ F allele carriers vs. carriers vs. Bt/Bt homozygotes, p<0.05; # bT allele carriers vs. Bt/Bt
f/f homozygotes, p<0.05 without correction homozygotes for dynamic torque-velocity curve, p=0.05
1240
The association between VDR genotypes and muscle
strength has already been studied in both men [5, 25, 28]
and women [23–27]. Unlike Dukas et al. [25], Grundberg et
al. [26], Geusens et al. [23, 24] and Wang et al. [27], we
found no association between BsmI or TaqI genotype and
hamstrings or quadriceps strength in our group of women.
Since these previous studies included either young, middle
aged or senior women, we repeated the analyses for the
middle-aged and senior women separately. Also within the
separate cohorts no association of quadriceps or hamstrings
strength with BsmI or TaqI was observed. However, an
association of the TaqI polymorphism with handgrip
strength was found in adult women (data not shown).
Several remarks should be given on this lack of
replication in our study. First, one should be careful in
comparing results from Asian [27] and Caucasian popula-
tions since VDR allele frequencies and linkage disequilib-
rium between markers are largely ethnicity-dependent. As a
consequence, even if the same allele is found to be
associated with a trait in both Caucasians and Asians, this
can be due to linkage to a completely different allele [31].
Uitterlinden et al. [31], therefore, suggested to analyse
haplotypes rather than individual polymorphisms, which we
have done in our study. Second, results from Geusens et al.
[23, 24] were adjusted for age, calcium intake and bone
mineral density. Unfortunately, we do not have data on
these potentially confounding factors for all of the subjects
and we therefore did not include them in our analyses.
To our knowledge, no previous studies have investigated
the effect of the FokI polymorphism on muscle strength in
women. We found significant differences between FokI
genotype groups for isometric and concentric quadriceps
strength. Adjustment for age, height and total fat-free mass
influenced the genotype-phenotype association in isometric
quadriceps strength. One possible explanation for this
confounding effect is the genotypic difference in height
observed in our sample: f/f homozygotes are taller than F
allele carriers and also have higher strength. Since peak
torque is dependent on the length of the lever arm, longer
legs can induce higher torque values.
We should point out that FokI genotype distribution was
not in Hardy-Weinberg equilibrium in our total sample of
women (p=0.011). While the consequences have been
widely addressed within a case-control approach (e.g.,
genotyping errors if controls are not in HWE), effects on
analyses of quantitative traits are far less obvious. However,
we believe that this disequilibrium was less likely a result
of genotyping errors and did not affect our results
substantially because: (1) The Sequenom MassARRAY
system is highly accurate; (2) FokI is a functional poly-
morphism that can influence VDR transcriptional activity
[32]. As a consequence selection can not be ruled out,
thereby violating one of the assumptions underlying the
Osteoporos Int (2007) 18:1235–1242
theory of HWE; (3) Deviation from HWE can occur when
subpopulations with different allele frequencies are present
within the population. In our cohorts, the middle-aged
compared to the senior women indeed had different allele
frequencies, but per group these allele frequencies were in
HWE.
Our data show that male bT haplotype carriers have less
handgrip and less concentric quadriceps strength than Bt
homozygotes. This contrasts with Van Pottelbergh et al. [5]
and Dukas et al. [25] who found no such association. As for
the FokI polymorphism, no association was detected in our
sample of men. These results are comparable with studies
from both Roth et al. [28] and Van Pottelbergh et al. [5].
Uitterlinden et al. [31] showed the importance of looking
at interaction effects between several polymorphisms within
the same gene. Fang et al. [13] also underline the
importance of studying both promoter and 3′-UTR poly-
morphisms because together they can determine how much
VDR mRNA will be expressed in a given target cell.
However, no interaction effects between the BsmI-TaqI
haplotype and the FokI polymorphism were found within
these samples. Probably this is due to the rather small
number of subjects in each interaction group, rendering our
study statistically underpowered to detect interactions.
Since we performed a rather large number of tests, one
could argue that a correction for multiple testing (e.g.,
Bonferroni correction) is appropriate. However, Bonferroni
correction is based on the number of independent test.
Given the biological interdependence of the different
muscle strength measurements it is difficult to assess the
exact number of independent test and therefore it is hard to
select the correction level. Moreover, this correction is
known to be overly conservative and would invalidate the
significance of the present results as well as those from
several previous studies investigating the association
between VDR polymorphisms and muscle strength.
By using multivariate models to analyse the torque-
velocity curves, different effects can be tested: (1) the main
effect of genotype by looking at the up- or down-ward shift
in the curve (intercept-effect) (2) the main effect of
contraction velocity by comparing the genotype-indepen-
dent strength levels over the different velocities and (3) the
interaction effect between genotype and velocity by testing
significance of shape differences of the torque-velocity
curve according to VDR genotype (slope effect). Our
results indicate both a genotype effect i.e., f/f homozygous
women and Bt/Bt homozygous men have higher strength
than F or bT allele carriers, respectively, and a velocity
effect i.e., lower strength levels at higher velocities. How-
ever, neither for men nor for women an interaction between
genotype and velocity was found, indicating that the
polymorphisms have a general strength increasing effect
rather than a velocity-dependent effect.
Osteoporos Int (2007) 18:1235–1242
The molecular basis for the associations observed for the
polymorphisms in the present study remains uncertain. The
BsmI and TaqI polymorphisms are located within the 3′-end
of the VDR gene and do not directly affect protein structure
or function. Several explanations for the associations have
been suggested. The 3′-UTR is known to have a regulatory
role in mRNA stability. The selected polymorphisms could
influence this stability and thereby regulate expression
either directly or through linkage disequilibrium with other
regulatory SNPs like e.g., the polyA repeat [33]. Whitfield
et al. [34], on the other hand, suggest that the translational
activity of the various mRNA variants could differ.
The FokI SNP, however, is one of the few functional
polymorphisms known in the VDR gene. Functional studies
have demonstrated that the T to C transition of this
polymorphism results in a 3 amino acids shorter protein,
with higher activity [32] because of a more efficient
interaction with transcription factor TFII B [35, 36].
Moreover, gene-specific effects could be present since
some promoter areas of the Vitamin D target genes may
be more sensitive to these VDR genotype-dependent
differences in activity than others [31]. It remains unclear
why gender-specific effects are observed both FokI and
BsmI and TaqI genotypes. Furthermore, comparisons with
other studies are difficult as most reports are limited to one
gender.
Vitamin D has long been known for its influence on
muscle tissue [37, 38]. However, the specific pathways
through which this occurs are less well known. Several
investigators suggest that Vitamin D could exert its actions
through the VDR-mediated regulation of insulin-like
growth factor binding proteins [39, 40], which in turn
regulate the actions of Insulin-Like Growth Factors [41],
known as modulators for muscle strength and function [40,
42–44].
Several limitations of this study should be addressed.
First, data were not available for all individuals regarding
Calcium intake, serum or plasma vitamin D levels, bone
mineral density or fracture risk. Therefore these could not
be included in the analyses and no direct assessment of the
relationship between muscle strength and these factors
could be made. It should, however, be pointed out that
subjects with joint replacements or drug treatments influ-
encing bone mineral density were excluded from the
analyses. Second, as mentioned earlier, sample sizes are
limited which implies that only large genotype effects could
be detected and power to detect allelic interaction effects
was insufficient. Third, since maximum torque at 180°/s
extension/flexion was determined within a 25 repetition test
at the end of the strength testing protocol, values are
probably lower than expected from the theoretical strength-
velocity curve [45]. Additionally, isometric values shown in
Figs. 1 and 2 are measured at 120° knee angle for knee
1241
flexion and extension. This is positioned at different points
in the force-length curve for knee flexor and extensor
muscles. Furthermore, dynamic torques were not registered
at this specific knee angle only, but were measured as the
peak value over the total range of motion (90°–160°). This
might also induce deviations from the typical shape of the
strength-velocity curve.
In conclusion, VDR FokI f/f homozygote women,
perform better in isometric quadriceps muscle strength
and show a trend towards an increased concentric torque-
velocity curve, compared to F allele carriers. In men, Bt/Bt
homozygotes for the BsmI-TaqI haplotype outperform bT
carriers in both isometric and concentric quadriceps torques
at different contraction velocities.
Acknowledgements Strength phenotyping of both middle-aged and
senior groups was supported by the Flemish Government in the Policy
Research Centre Sport, Physical Activity and Health. Genotyping was
supported by a Research grant to Martine Thomis of the Fund for
Scientific Research Flanders (FWO). Gunther De Mars is funded by
grant G.0496.05 of the FWO. An Windelinckx is funded by the
Research Fund of the K.U.Leuven (OT/04/44).
References
1. Kannus P, Uusi-Rasi K, Palvanen M et al (2005) Non-pharmaco-
logical means to prevent fractures among older adults. Ann Med
37(4):303–310
2. Morrison NA, Yeoman R, Kelly PJ et al (1992) Contribution of
trans-acting factor alleles to normal physiological variability:
vitamin D receptor gene polymorphisms and circulating osteocal-
cin. Proc Natl Acad Sci USA 89(15):6665–6669
3. Morrison NA, Qi JC, Tokita A et al (1994) Prediction of bone density
from vitamin D receptor alleles. Nature 367(6460):284–287
4. Kikuchi R, Uemura T, Gorai I et al (1999) Early and late
postmenopausal bone loss is associated with BsmI vitamin D
receptor gene polymorphism in Japanese women. Calcif Tissue Int
64(2):102–106
5. Van Pottelbergh I, Goemaere S, De Bacquer D et al (2002)
Vitamin D receptor gene allelic variants, bone density, and bone
turnover in community-dwelling men. Bone 31(5):631–637
6. Duman BS, Tanakol R, Erensoy N et al (2004) Vitamin D receptor
alleles, bone mineral density and turnover in postmenopausal
osteoporotic and healthy women. Med Princ Pract 13(5):260–266
7. Remes T, Vaisanen SB, Mahonen A et al (2005) Bone mineral
density, body height, and vitamin D receptor gene polymorphism
in middle-aged men. Ann Med 37(5):383–392
8. Uitterlinden AG, Pols HA, Burger H et al (1996) A large-scale
population-based study of the association of vitamin D receptor
gene polymorphisms with bone mineral density. J Bone Miner Res
11(9):1241–1248
9. Aerssens J, Dequeker J, Peeters J et al (2000) Polymorphisms of
the VDR, ER and COLIA1 genes and osteoporotic hip fracture in
elderly postmenopausal women. Osteoporos Int 11(7):583–591
10. van der Sluis I, de Muinck Keizer-Schrama SM, Krenning EP et al
(2003) Vitamin D receptor gene polymorphism predicts height
and bone size, rather than bone density in children and young
adults. Calcif Tissue Int 73(4):332–338
11. Dvornyk V, Liu PY, Long JR et al (2005) Contribution of
genotype and ethnicity to bone mineral density variation in
1242
Caucasians and Chinese: a test for five candidate genes for bone
mass. Chin Med J (Engl ) 118(15):1235–1244
12. Macdonald HM, McGuigan FE, Stewart A et al (2006) Large-
scale population-^based study shows no evidence of association
between common polymorphism of the VDR gene and BMD in
British women. J Bone Miner Res 21(1):151–162
13. Fang Y, van Meurs JB, d’Alesio A et al (2005) Promoter and 3′-
untranslated-region haplotypes in the vitamin D receptor gene
predispose to osteoporotic fracture: the Rotterdam study. Am J
Hum Genet 77(5):807–823
14. Garnero P, Munoz F, Borel O et al (2005) Vitamin D receptor gene
polymorphisms are associated with the risk of fractures in
postmenopausal women, independently of bone mineral density.
J Clin Endocrinol Metab 90(8):4829–4835
15. Nguyen TV, Esteban LM, White CP et al (2005) Contribution of
the collagen I alpha1 and vitamin D receptor genes to the risk of
hip fracture in elderly women. J Clin Endocrinol Metab 90
(12):6575–6579
16. Sipila S, Heikkinen E, Cheng S et al (2006) Endogenous hormones,
muscle strength, and risk of fall-related fractures in older women. J
Gerontol A Biol Sci Med Sci 61(1):92–96
17. Lord SR, Ward JA, Williams P et al (1994) Physiological factors
associated with falls in older community-dwelling women. J Am
Geriatr Soc 42(10):1110–1117
18. Stel VS, Smit JH, Pluijm SM et al (2003) Balance and mobility
performance as treatable risk factors for recurrent falling in older
persons. J Clin Epidemiol 56(7):659–668
19. Luukinen H, Koski K, Laippala P et al (1997) Factors predicting
fractures during falling impacts among home-dwelling older
adults. J Am Geriatr Soc 45(11):1302–1309
20. Nguyen ND, Pongchaiyakul C, Center JR et al (2005) Identifica-
tion of high-risk individuals for hip fracture: a 14-year prospective
study. J Bone Miner Res 20(11):1921–1928
21. Robbins JA, Schott AM, Garnero P et al (2005) Risk factors for hip
fracture in women with high BMD: EPIDOS study. Osteoporos Int
16(2):149–154
22. Bischoff HA, Borchers M, Gudat F et al (2001) In situ detection
of 1,25-dihydroxyvitamin D3 receptor in human skeletal muscle
tissue. Histochem J 33(1):19–24
23. Geusens P, Vandevyver C, Vanhoof J et al (1997) Quadriceps and
grip strength are related to vitamin D receptor genotype in elderly
nonobese women. J Bone Miner Res 12(12):2082–2088
24. Vandevyver C, Vanhoof J, Declerck K et al (1999) Lack of
association between estrogen receptor genotypes and bone mineral
density, fracture history, or muscle strength in elderly women. J
Bone Miner Res 14(9):1576–1582
25. Dukas L, Bischoff H, Michiels L et al (2001) Vitamin D receptor
genotype in community-dwelling elderly men and women: its
relation to muscle strength results from the cross-sectional
analyses of the baseline measurements of the AIMS-study.
Abstract book of the 23rd annual meeting of the American
Society of Bone and Mineral Research, S353
26. Grundberg E, Brandstrom H, Ribom EL et al (2004) Genetic
variation in the human vitamin D receptor is associated with
muscle strength, fat mass and body weight in Swedish women.
Eur J Endocrinol 150(3):323–328
27. Wang P, Ma LH, Wang HY et al (2006) Association between
polymorphisms of vitamin D receptor gene ApaI, BsmI and TaqI
Osteoporos Int (2007) 18:1235–1242
and muscular strength in young Chinese women. Int J Sports Med
27(3):182–186
28. Roth SM, Zmuda JM, Cauley JA et al (2004) Vitamin D receptor
genotype is associated with fat-free mass and sarcopenia in elderly
men. J Gerontol A Biol Sci Med Sci 59(1):B10–B15
29. Claessens AL, Beunen G, Malina RM (2000) Anthropometry,
physique, body composition and maturity. In: Armstrong N, van
Mechelen W (eds) Paediatric exercise science and medicine.
Oxford University Press, Oxford, pp 11–22
30. Durnin JVGA, Womersley J (1974) Body fat assessment from
total body density and its estimations from skinfold thickness:
measurements on 481 men and women aged from 16 to 72 years.
Br J Nutr 32:77–87
31. Uitterlinden AG, Fang Y, van Meurs JBJ et al (2004) Genetics and
biology of vitamin D receptor polymorphisms. Gene 338(2):143–156
32. Arai H, Miyamoto K, Taketani Y et al (1997) A vitamin D
receptor gene polymorphism in the translation initiation codon:
effect on protein activity and relation to bone mineral density in
Japanese women. J Bone Miner Res 12(6):915–921
33. Gennari L, Becherini L, Falchetti A et al (2002) Genetics of
osteoporosis: role of steroid hormone receptor gene polymor-
phisms. J Steroid Biochem Mol Biol 81(1):1–24
34. Whitfield GK, Remus LS, Jurutka PW et al (2001) Functionally
relevant polymorphisms in the human nuclear vitamin D receptor
gene. Mol Cell Endocrinol 177(1–2):145–159
35. Jurutka PW, Remus LS, Whitfield GK et al (2000) The polymorphic
N terminus in human vitamin D receptor isoforms influences
transcriptional activity by modulating interaction with transcription
factor IIB. Mol Endocrinol 14(3):401–420
36. Colin EM, Weel AE, Uitterlinden AG et al (2000) Consequences
of vitamin D receptor gene polymorphisms for growth inhibition
of cultured human peripheral blood mononuclear cells by 1, 25-
dihydroxyvitamin D3. Clin Endocrinol (Oxf) 52(2):211–216
37. Pfeifer M, Begerow B, Minne HW (2002) Vitamin D and muscle
Function. Osteoporos Int 13(3):187–194
38. Montero-Odasso M, Duque G (2005) Vitamin D in the aging
musculoskeletal system: An authentic strength preserving hor-
mone. Mol Aspects Med 26(3):203–219
39. Peng L, Malloy PJ, Feldman D (2004) Identification of a
functional vitamin D response element in the human insulin-like
growth factor binding protein-3 promoter. Mol Endocrinol 18
(5):1109–1119
40. Sweeney C, Murtaugh MA, Baumgartner KB et al (2005) Insulin-
like growth factor pathway polymorphisms associated with body
size in Hispanic and non-Hispanic white women. Cancer Epidemiol
Biomarkers Prev 14(7):1802–1809
41. Jones JI, Clemmons DR (1995) Insulin-like growth factors and
their binding proteins: biological actions. Endocr Rev 16(1):3–34
42. Florini JR, Ewton DZ, Coolican SA (1996) Growth hormone and
the insulin-like growth factor system in myogenesis. Endocr Rev
17(5):481–517
43. Noguchi S (2005) The biological function of insulin-like growth
factor-I in myogenesis and its therapeutic effect on muscular
dystrophy. Acta Myol 24(2):115–118
44. Glass DJ (2005) Skeletal muscle hypertrophy and atrophy
signaling pathways. Int J Biochem Cell Biol 37(10):1974–1984
45. Hill A (1938) The heat of shortening and the dynamic constants of
muscle. Proc R Soc Lond B Biol Sci 126(843):136–195