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Experimental Gerontology 48 (2013) 795800

Contents lists available at SciVerse ScienceDirect

Experimental Gerontology
journal homepage: www.elsevier.com/locate/expgero

Metabolic changes in the auditory cortex in presbycusis demonstrated

by MR spectroscopy
Oliver Profant a, c,, Zuzana Balogov a, c, Monika Dezortov b, Dita Wagnerov b, Milan Hjek b, Josef Syka a
a
b
c

Department of Auditory Neuroscience, Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, Prague, Czech Republic
MR Unit, Department of Diagnostic and Interventional Radiology, Institute for Clinical and Experimental Medicine, Prague, Czech Republic
Department of Otorhinolaryngology and Head and Neck Surgery, 1st Faculty of Medicine, Charles University, University Hospital Motol, Prague, Czech Republic

a r t i c l e

i n f o

Article history:
Received 10 January 2013
Received in revised form 4 April 2013
Accepted 25 April 2013
Available online 4 May 2013
Section Editor: Christian Humpel
Keywords:
Presbycusis
Auditory cortex
MR spectroscopy
Glutamate
N-acetylaspartate
Lactate
GABA

a b s t r a c t
In humans, aging is accompanied by the deterioration of the hearing function presbycusis. The major etiology for
presbycusis is the loss of hair cells in the inner ear; less well known are changes in the central auditory system.
Therefore, we used 1 H magnetic resonance spectroscopy at 3 T tomograph to examine metabolite levels in the
auditory cortex of three groups of subjects: young healthy subjects less than 30 years old and subjects older than
65 years either with mild presbycusis corresponding to their age or with expressed presbycusis. Hearing function
in all subjects was examined by pure tone audiometry (12516000 Hz). Signicant differences were found in the
concentrations of glutamate and N-acetylaspartate, with lower levels in aged subjects. Lactate was particularly
increased in subjects with expressed presbycusis. Signicant differences were not found in other metabolites,
including GABA, between young and elderly subjects. The results demonstrate that the age-related changes of
the inner ear are accompanied by a decrease in the excitatory neurotransmitter glutamate as well as a lactate
increase in the auditory cortex that is more expressed in elderly subjects with large hearing threshold shifts.
2013 Elsevier Inc. All rights reserved.

1. Introduction
Physiological aging is accompanied by a deterioration of sensory and
cognitive functions. A highly prevalent sensory decline is the sensorineural hearing loss called presbycusis, which starts to affect the middle
aged human population or as they grow older. Accumulated noise exposure, cardiovascular disorders and their treatment, as well as hereditary
susceptibility are several factors that inuence the state of hearing in
the elderly. Overall, hearing thresholds deteriorate on average by 1 dB
per year for persons over 60, depending on gender and initial thresholds
(Davis, 1991 and Gates and Cooper, 1991). Presbycusis in humans
causes a deterioration of speech comprehension, especially in the presence of background noise, which leads to poor communication and results in social deprivation (Sprinzl and Riechelmann, 2010).
For many years, disorders of the inner ear were considered as the
primary cause of presbycusis. The loss of functional outer hair cells,
the loss of afferent neurons in the cochlea, disorders of the stria vascularis
and stiffening of the basilar membrane were found to be the main reasons
Abbreviations: EP, elderly subjects with expressed presbycusis; MP, elderly subjects
with mild presbycusis; NAA, N-acetylaspartate; VOI, volume of interest; YC, young subjects with physiologic hearing.
Corresponding author at: Department of ENT, Charles University in Prague, 1st Faculty of
Medicine, University Hospital Motol, V valu 84, 150 06 Prague 5, Czech Republic.
Tel.: +420 224434301, +420 224434358; fax: +420 224434310.
E-mail address: profant@biomed.cas.cz (O. Profant).
0531-5565/$ see front matter 2013 Elsevier Inc. All rights reserved.
http://dx.doi.org/10.1016/j.exger.2013.04.012

for the peripheral component of presbycusis (Schuknecht and Gacek,

1993). Typically, presbycusis shows a decline in frequency audibility towards high frequencies, which are important for speech recognition.
Decreased speech understanding, as well as the altered processing
of the temporal parameters of acoustical signals and intensity discrimination, point towards a possible central component of presbycusis
(Gordonsalant and Fitzgibbons, 1993; Gordon-Salant et al., 2010;
Mazelov et al., 2003; Moore et al., 1992; Snell and Frisina, 2000). The
occurrence of central components of presbycusis in humans, besides
audiological testing, has been mostly demonstrated by functional magnetic resonance imaging (fMRI). In the elderly, the loss of sensory activation of the auditory cortex is accompanied by increased activity in the
cognitive regions of the prefrontal cortex (Wong et al., 2008). The increased activity of the prefrontal cortex in the aging population has
been further positively correlated with an improved ability to understand
speech in noise (Akeroyd, 2008; Wong et al., 2010).
Experiments in animals revealed that aging is accompanied by significant changes in the structure and function of the auditory system.
Besides changes in the electrophysiological parameters, the most
expressed age-related changes are those in brain neurochemistry. Both
in the subcortical nuclei and the auditory cortex of rats, a decreased
presence of the inhibitory neurotransmitter GABA was found with
aging using immunocytochemical methods (Burianova et al., 2009;
Caspary et al., 1990; Syka, 2010) as well as changes in calcium binding proteins (Idrizbegovic et al., 2004; Ouda et al., 2008; Zettel et al.,

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O. Profant et al. / Experimental Gerontology 48 (2013) 795800

1997; Zettel et al., 2003). The results of experiments in animals demonstrated the disorganization of the system of neurotransmitters
and calcium regulatory proteins in the central auditory system in
presbycusis, and therefore it can be assumed that a similar decrease
of GABA and a further disorganization of brain neurochemistry may
exist in man.
Several studies concerning age-related differences in the concentration of brain metabolites were performed using 1 H magnetic
resonance spectroscopy (MRS) as early as in the nineties (Kreis,
1997), at that time with 1.5 T MR systems. For example, Saunders et
al. (1999) and Leary et al. (2000) did not nd any differences in the
tissue concentrations of major metabolites such as N-acetylaspartate
(NAA), total creatine, choline compounds and myo-inositol between
young and old subjects. A small age-related increase in creatine concentration was observed by Saunders et al. (1999) and Leary et al. (2000) in
the white matter of the parietal lobe. Similarly, Pfefferbaum et al.
(1999) reported increased concentrations of creatine and choline in
older subjects in comparison with young subjects, with the concentrations of other metabolites remaining unchanged. Schuff et al. (2001)
studied changes of the major metabolites in the white matter and observed increases in creatine and NAA when comparing 50-year-old
with 90-year-old subjects. In their study, Ferguson et al. (2002) concluded
that in healthy humans aged 6570 years, metabolite levels correspond
to cognitive performance. However, the low sensitivity of 1.5-T systems
in determining the concentrations of glutamate, glutamine and GABA
did not allow information to be obtained at that time about the levels of
these metabolites, which represent major brain transmitters.
With advances in MR technology, more recent studies mostly
performed on 3-T or higher magnetic eld systems allowed the more precise investigation of neurotransmitters in the brain (Nacewicz et al., 2012;
O'Gorman et al., 2011; Schubert et al., 2004; Stone, 2009; Waddell et al.,
2011). Boumezbeur et al. (2010) used 13 C/1 H MR spectroscopy to
quantitatively characterize the rates of neuronal and astroglial tricarboxylic acid cycles and neuroglial glutamateglutamine cycling in healthy elderly and young subjects. Compared with young subjects, neuronal
mitochondrial metabolism and glutamateglutamine cycle metabolism
ux were 30% lower in elderly subjects.
However, information about the changes in the concentrations of metabolites in the auditory system in humans, particularly in the auditory
cortex, similar to those found in experimental animals, is still missing.
Therefore, 1 H MRS was used in our experiments with a 3 T system as a
noninvasive tool to determine the levels of various metabolites, including
neurotransmitters, in the auditory cortex of young healthy controls and
elderly subjects with and without expressed presbycusis.
2. Material and methods
2.1. Subjects
Fifty-three subjects underwent audiological and MRS examinations.
They were divided into three groups based on the audiological results:
17 elderly subjects with expressed presbycusis (EP; 8 females/9
males, mean age 71.2 5.2 year), 18 elderly controls age-matched to
the EP group with a mild degree of presbycusis (MP; 13 females/5
males, mean age 67.3 1.9 year) and 18 young controls with normal
hearing not exceeding 20 dB HL (YC; 7 females/11 males, mean age
24.4 2.3 year). None of the examined patients underwent any
previous otological surgery nor did they suffer from a vestibular lesion,
tinnitus, severe head trauma, lesions of the facial nerve, disorders of the
cervical spine, and self-reported central nervous system disorders or
any contraindication for safe MRI scanning. None of the subjects were
musical professionals, but several in both of the elderly groups played
musical instruments regularly during their youth. All of the examined
subjects were informed about the examination protocol, and their
written consent was obtained according to the local Ethical Committee
rules.

2.2. Assessment of auditory function

An otoscopic examination with the removal of the cerumen and
conrmation of an intact tympanic membrane were performed in all
subjects. For assessing the hearing abilities of all three subject groups
pure tone audiometry from 125 to 16,000 Hz and tympanometry
were used. Audiometric investigations were performed in a sound
attenuated chamber; all measurements were performed monaurally,
and the ears of each subject were tested successively.
Tympanometry was performed with an Interacoustics AZ26
tympanometer to conrm optimal middle ear conditions and an intact tympanic membrane. For pure tone audiometry over an extended
frequency range from 125 Hz to 16 kHz, a Madsen Orbiter 922, version
2, audiometer was used, calibrated by the Czech Institute of Metrology
according to the Czech State Norm and the European Standards EN ISO
389-1 and EN ISO 389-5. Acoustical signals were delivered monaurally
via Sennheiser HAD 200 high-frequency headphones. Audiograms were
measured in one-octave steps at frequencies ranging from 125 Hz to
8 kHz and then at 10, 12.5 and 16 kHz. Hearing thresholds were detected
with a resolution of 5 dB steps; the contralateral ear was masked if
necessary. Bone conduction testing (frequencies in the range of 250
4000 Hz) was performed to exclude possible elevated thresholds due to
conductive hearing loss.
On the basis of the recorded hearing thresholds and in comparison
with the ISO 7029 norm specied for their age (at frequencies of 2, 4
and 8 kHz), the elderly patients were divided into two groups. In
order to be certain that the group elderly with expressed presbycusis
(EP) had markedly worse hearing than the group elderly with mild
presbycusis (MP), only subjects with a hearing threshold elevation
(at least at 2 frequencies) categorized as being among the worst 25%
of the population according to the ISO 7029 norm specied for their
age and gender were used.
2.3. MR examination
All subjects underwent a 1 H MRS examination performed at 3 T in a
whole-body Siemens TRIO system using a Tx/Rx head coil. Base images
by T2-weighted turbo spin echo sequences in three perpendicular
planes (TR/TE = 4400/99 ms, 4 mm slice thickness) were obtained
for spectra localization. A MPRage sequence in the sagittal plane with
1 mm slice thickness was used for segmentation, and white matter
(WM), gray matter (GM) and cerebrospinal uid (CSF) contents were
estimated in all spectroscopic regions of interest. MR spectroscopy
was performed by a single voxel technique (PRESS: TR/TE = 5000/
30 ms, 64 acquisitions); the volumes of interest (VOI 8 and 18 ml)
were centered on the Heschl gyrus in both temporal lobes (Fig. 1). For
quantication, spectra without water suppression with one acquisition
were also obtained.
MRS data were analyzed by the LCModel (Provencher, 1993) with a
database of 14 metabolites including macromolecules and lipids; the
water signal was used as an internal standard for quantication. CSF
corrections from the segmentation were applied.
WM, GM and CSF contents in the voxel were estimated as follows:
a high-resolution 3D structural MR dataset was acquired by means of
a MPRage sequence. This dataset was segmented by SPM8 software
(Ashburner and Friston, 2005), (www.l.ion.ucl.ac.uk/spm/) to acquire probability maps of GM, WM and CSF in the whole brain region.
Data corresponding to the region of the measured spectroscopic voxel
were extracted from the maps by means of self-written software in
Java language. Position information and orientation information in structural and spectroscopic datasets were used for the spatial co-registration
of probability maps with single voxel MRS data. By averaging the
extracted probability maps, the proportions of GM, WM and CSF in a
particular spectroscopic voxel were computed.
The statistical evaluations were performed by one way ANOVA
with Tukey's multiple comparison test using GraphPad Prism software

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O. Profant et al. / Experimental Gerontology 48 (2013) 795800

797

(www.graphpad.com). Data for ANOVA analysis were prepared in Excel,

and three groups of subjects (YC, MP and EP), each with 18 concentrations of metabolites, were compared (ANOVA df (2, 103); a signicant
difference was considered to be p b 0.05).
3. Results
3.1. Investigation of hearing function
The control group of young subjects (YC) displayed normal hearing with thresholds not exceeding 20 dB HL above the values typical
for their age (Fig. 2). The MP group showed physiological auditory
thresholds ( 20 dB HL) with a small but steady increase of thresholds at frequencies above 3 kHz with a 69 dB hearing loss at 12 kHz
and the absence of hearing at 16 kHz. In the EP group the threshold
increase exceeding the physiological range started above 750 Hz
and reached a 76.5 dB hearing loss at 12.5 kHz with no hearing at
16 kHz. The hearing thresholds of all three groups were statistically
compared at frequencies above 750 Hz, where the EP group's average
threshold started to exceed the physiological hearing range (20 dB
HL). The YC group's thresholds were signicantly lower than those of
the MP group and also signicantly lower than those of the EP subjects
(Tukey's multiple comparison test, F (2, 27) = 16.93; p = 0.016 for
MP and p b 0.0001 for EP); the hearing thresholds of both elderly groups
signicantly differed between each other as well (Tukey's multiple
comparison test, F (2, 27) = 16.93; p = 0.025). No signicant difference
between the left and right ears was found, therefore the thresholds from
both ears were merged together. Tympanometric examination showed
type A curve in all volunteers.

No signicant differences were found between the metabolite concentrations from the left and right hemispheres, therefore the MRS results were pooled from both hemispheres for both the 8 ml as well as
the 18 ml VOI. Left-handedness or gender differences also did not affect
the results.
In both groups of elderly subjects, a certain degree of brain atrophy was observed on MR images. Therefore, segmentation of the
CSF volume was necessary for spectra quantication. Independent
of VOI size, CSF content inside the VOI was comparable in both elderly groups and was 78% higher than in young controls. A comparison
of the brain tissue compositions in the VOI showed similar amounts
of WM as compared to GM among the subject groups; nevertheless,
the contents differed between the small and large volumes, see
Table 1.
Graphs of metabolite concentrations are shown in Fig. 3 and Table 1
for all the examined groups of subjects separately for 8 and 18 ml VOI.
Data were compared between the YC and both aged groups as well as
between MP and EP and statistically analyzed by one way ANOVA
(Tukey's multiple comparison test). In both elderly groups the concentration of N-acetylaspartate (including N-acetylaspartylglutamate) was
signicantly lower (for 8 ml VOI: F (2, 103) = 8.525, p = 0.0078 for YC
vs. MP, p = 0.0005 for YC vs. EP; and for 18 ml VOI: F (2, 103) = 8.525,
p = 0.0022 for YC vs. MP, p b 0.0001 for YC vs. EP) in comparison with
young controls, independent of the VOI size. No differences were found
between the MP and EP groups in this case. Similarly, the glutamate
concentration was lower in elderly subjects when compared with
young subjects (for 18 ml VOI: F (2, 103) = 28.37, p b 0.0001 for YC
vs. MP, p b 0.0001 for YC vs. EP); however, in this case the difference
between young subjects and elderly controls (YC and MP) did not
reach statistical signicance in the smaller (8 ml) VOI. Glutamine itself
did not reveal any differences.
Signicant differences between all individual groups were found in
lactate concentration measured from the large VOI (for 18 ml VOI: F
(2, 103) = 13.57, p b 0.0001 for YC vs. MP, p b 0.0143 for YC vs. EP,

Fig. 1. Example of spectra localization (small 8 ml VOI in black and large 18 ml VOI in white)
in the temporal lobes of both hemispheres of one subject centered around the Heschl gyrus.

p = 0.0493 for MP vs. EP), with higher values in MP than in YC and

even higher values in EP than in MP. This trend was also visible in the
small VOI but without statistical signicance. Other statistically signicant changes were found in decreasing creatine with phosphocreatine
(Cr/PCr) (for 18 ml VOI: F (2, 103) = 4.708, p = 0.0131 for YC vs. EP)
and increasing GABA in the EP group compared to YC only in the case
of the large VOI (for 18 ml VOI: F (2, 103) = 3.847, p = 0.0413 for
YC vs. EP). No signicant age-related changes were found in myoinositol or in choline compounds in either VOI.

Hearing threshold in all three groups

0.25

0.75

1.5

10 16

0
YC

10

hearing loss [dB HL]

3.2. MR results

20

MP

30

EP

40

mean

50

median

60
70
80
90
100
0.125

0.5

8 12.5

frequency [kHz]
Fig. 2. Hearing thresholds of subjects in the YC, MP and EP groups: the results of pure tone
audiometry in an extended high-frequency range. Data are shown as means (SEM) and
medians (in the case of no hearing sensation, the highest intensity of the stimulus was set
as the threshold). An absence of hearing at 16 kHz was found in the MP and EP groups,
therefore dashed lines were used. Statistical analysis (one-way ANOVA, Tukey's multiple
comparison test) was used to compare the hearing thresholds above 750 Hz (dashed
area), * p b 0.05 (not shown is the comparison of YC vs. EP, p b 0.0001). At frequencies
above 750 Hz, the EP group exceeded the physiological threshold set at 20 dB HL.

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O. Profant et al. / Experimental Gerontology 48 (2013) 795800

Table 1
Metabolite concentrations [mM] and white matter and gray matter content [%] with their standard deviations in the brain tissue of the auditory cortex calculated by the LCModel
technique with segmentation to CSF content (GABA -aminobutyrate, Gln glutamine, Glu glutamate, Ins myo-inositol, Lac lactate, Cho choline compounds, NAA
N-acetylaspartate + N-acetylaspartylglutamate, Cr/PCr creatine + phosphocreatine, WM white matter, and GM gray matter).
Group
VOI 8 ml
YC
PH
EP

VOI 18 ml
YC
PH
EC

GABA

Gln

Glu

Ins

Lac

Cho

NAA

Cr/PCr

Glx

WM

GM

1.77
0.9
1.56
0.8
1.79
0.8

4.05
2
3.57
2.4
4.23
1.7

14.27
2
13.08
2
11.80

7.74
1
8.57
1.3
8
1.7

0.74
0.6
1.04
0.9
1.26
1

2.2
0.3
2.33
0.4
2.16
0.4

12.3
0.8
11.14
1.8
10.79

10.19
0.8
10.2
1.3
9.83
1.4

18.33
3.4
16.64
3.2
16.03
3.2

22
5
23.8
4
23.5
3.5

78
5
76.2
4
76.5
3.5

1.22
0.7
1.61
0.7
1.64
0.7

1.76
1.1
2.04
1.7
1.7
1

12.1
1.4
10.14

7.59
0.9
7.68
1
7.41
1.5

0.53
0.3
0.84#
0.5
1.10
0.5

2
0.3
2.15
0.3
1.95
0.4

11.32
1.3
9.97
1.8
9.49
1.8

13.86
1.9
12.17
2.9
11.13
2

26.7
4.6
30
3.5
28.4
3.3

73.3
4.6
70
3.5
71.6
3.3

2.6

1.7
9.43
1.5

9.54
0.8
9.4
1
8.77
1.4

p b 0.05.
p b 0.01.
p b 0.001 from YC.
#
p b 0.05 from EP.

4. Discussion
The results of our MRS investigation demonstrate that differences
exist in the concentration of metabolites in the auditory cortex between
young and elderly subjects. Signicantly lower levels of glutamate and
N-acetyl aspartate and higher levels of lactate are present in elderly
subjects with practically zero differences between subjects with mild
presbycusis and those with expressed presbycusis.

A
[mM]
18

***

***
***

16
14
12
10
8
6
4
2
0

GABA

Gln

Glu

Ins

Lac

Cho

NAA

Cr/PCr

***
**

NAA

Cr/PCr

B
[mM]
18
16
***
***

14
12
10
8
6
4

***
* *

2
0

GABA

Gln

Glu

Ins
YC

Lac
PH

Cho
EP

Fig. 3. Graphs of metabolite concentrations [in mM] and their standard deviations
in the auditory cortex. A. VOI 8 ml, and B. VOI 18 ml. (GABA -aminobutyrate,
Gln glutamine, Glu glutamate, Ins myo-inositol, Lac lactate, Cho choline
compounds, NAA N-acetylaspartate + N-acetylaspartylglutamate, and Cr/PCr
creatine + phosphocreatine). * p b 0.05, ** p b 0.01, and *** p b 0.001.

The effect of aging on the metabolite concentrations observable in

the human brain by MRS can be seen mainly in the NAA concentration
decline as NAA reects neuronal damage and demyelinating processes
and can be used as an indirect measure of neuronal integrity and density (Li et al., 2010; Richards et al., 1997; van Wageningen et al., 2010).
The decreased NAA levels found in both our aged groups conrm this
age-dependent neuronal degradation and are in agreement with other
studies at 3 T that revealed that NAA concentration decreases with
aging in many parts of the brain (Kaiser et al., 2005; Schubert et al.,
2004). In contrast to these ndings, higher NAA levels in the auditory
cortex were reported in musicians (Aydin et al., 2005) as a result of
long-lasting musical training, which is consistent with the increase of
synaptic density in use-dependent adaptation. However, this effect
was eliminated by the selection of our subjects.
In the case of metabolites such as creatine (with one small exception),
choline and myo-inositol, our examinations did not reveal any signicant
changes with aging. Although some limited data exist in the literature
about age-related changes of these metabolites (e.g., Leary et al., 2000,
reported an increase of creatine with aging), most results support our
data, i.e. there are no age-related changes in the concentrations of these
metabolites.
An age-related decrease in glutamate was found in the motor cortex
(Kaiser et al., 2005) and in the parietal gray matter and basal ganglia
and to a lesser degree in the frontal white matter (Chang et al., 2009).
It is therefore not surprising that in the VOI representing the auditory
cortex, we found lower concentrations of glutamate in the elderly,
especially in those suffering from more expressed presbycusis. A
lower concentration of glutamate was also found in EP subjects in
comparison with MP subjects, but the difference was not signicant.
In the larger VOI both MP subjects and EP subjects showed a highly
signicant decrease in glutamate concentration when compared
with young subjects. This nding corresponds with the worse hearing thresholds in these subjects and deteriorated speech discrimination
(Profant et al., 2012). A similar tendency was also reported by Schubert
et al. (2004), who measured the concentration of glutamate using 3 T
MR spectroscopy in the anterior cingulate cortex and the hippocampus
and compared the values obtained in young and old subjects. The concentration values and the variability obtained in our experiments, as well as
in theirs, are comparable when taking into account the correction for CSF:
with aging, the values signicantly decreased (p b 0.04). Interestingly, in
the rst whole genome association study for age-related hearing impairment (Friedman et al., 2009; Newman et al., 2012) the authors identied
a highly signicant and replicated single nucleotide polymorphism located at GRM7, the gene encoding metabotropic glutamate receptor type 7.

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O. Profant et al. / Experimental Gerontology 48 (2013) 795800

To what extent this nding can be related to the lower levels of the excitatory transmitter glutamate observed in elderly subjects remains to be
elucidated.
Glutamine is a product of glutamate metabolism and in MR spectra
is hard to distinguish from the glutamate peaks. A similar problem
exists for GABA signals. The reason for these difculties is the low
concentration of these metabolites and the multiplicity and overlap of
glutamine, glutamate and GABA signals, which result in insufcient
resolution for their precise quantitative estimation in vivo even with a
3 T system. Although the mean concentrations of GABA from our
measurements, between 1.2 and 1.8 mM, are comparable with other
published data (Jensen et al., 2005; O'Gorman et al., 2011; Waddell et
al., 2011), technical difculties do not allow measuring GABA concentration with sufcient precision. Thus, our hypothesis based on the results of animal experiments (Burianova et al., 2009; Ling et al., 2005),
i.e. that in the human auditory cortex we should nd an insufcient
function of inhibitory neurotransmitters expressed as a lower concentration of GABA, was not proven by the results. The level of GABA was
found to be even slightly higher in the EP group of subjects than in
the young subjects when the larger VOI was used.
Lactate concentration under physiological conditions is very low,
and thus it is usually only detected under pathological conditions,
when energy metabolism is affected. In our study, a signicant increase
in lactate concentration was found in the 18 ml VOI in those subjects
with mild presbycusis as well as those with expressed presbycusis. In
the brain, lactate is processed in the TCA cycle in mitochondria and
therefore acts as an energy source, used especially in strenuous situations (Ross et al., 2010). Our results conrm the previously reported
elevation of lactate in aged brain (Soros et al., 2006), which seems to
be a result of altered mitochondrial metabolism (Ross et al., 2010). In
one of the rst papers using MR spectroscopy for measuring metabolite
concentrations in the auditory cortex (Richards et al., 1997), the authors
reported that a lower MR spectroscopic lactate signal was observed in
healthy control subjects during a musical stimulus when compared to
scanner noise or the sound of sirens. Recently, small but signicant
increases of lactate in the human visual cortex induced by visual stimulation were detected at 7 T (Lin et al., 2012). A lactate increase in the
visual cortex was also consistently found using 1-H MRS in migraine
patients (Reyngoudt et al., 2012). Our nding that in the 18 ml VOI
the lactate levels in the AC were signicantly increased particularly
when comparing old subjects with expressed presbycusis with young
subjects, corresponds with the results of Emir et al. (2011), who found
that the lactate concentration in the visual cortex was signicantly
higher in elderly than in young subjects. The higher concentration of
lactate was accompanied in their measurements by a lower concentration of glutathione. The authors theorized that the lower glutathione and higher lactate concentrations are indications of defective
protection against oxidative damage and impaired mitochondrial
respiration. The difference in lactate levels between YC on one side
and MP and EP on the other side must be considered with caution
since the calculation of metabolite concentrations around 1 mM
using MRS is affected by a higher calculation error (CramerRao
bound > 50%). This fact can explain why we did not observe signicantly increased lactate levels in the smaller VOI, but only a tendency
towards an increase with aging.
In principle, the observed differences in metabolite concentrations
were more expressed in the larger VOI. Several reasons for this nding
can be found. One of these is the fact that the larger VOI contains
proportionally more white matter. The results of segmentation analysis
show that the presence of white matter was 37% higher than in the
smaller VOI. Therefore, the contribution of metabolites with a typically
higher concentration in the white matter compared to the gray matter
is somewhat greater. Another explanation can be found in the higher
S/N in the larger VOI, which was about twice as high as in the small
VOI; this plays an important role in the precision of the metabolite
quantication.

799

The atrophy of the aging brain as evident from the segmentation

analysis does not inuence the calculation of metabolite concentrations in the pure brain tissue. We found the same relative content of
WM and GM in all subject groups (in both VOI), which means that
the metabolic changes in the elderly groups of subjects depend on
aging and presbycusis only. Nevertheless, it should be highlighted
that the relaxation times could play an important role in the quantication of metabolites. Unfortunately, the age dependence of relaxation times has not been well described, especially in old age,
according to our knowledge, thus we did not include these corrections in our study.
As stated before, the aging of the auditory system is a complex
process, which comprises changes of the peripheral as well as the
central parts of the auditory system; however, it is unclear how
much these changes affect each other. Therefore, several hypotheses,
e.g. direct central presbycusis, indirect central presbycusis and
the cognitive hypothesis, have appeared to explain this complexity
(Humes et al., 2012; Tun et al., 2012). Peripheral changes that occur in
the organ of Corti and at the level of the spiral ganglion (Bao and
Ohlemiller, 2010) are accompanied by changes in the cochlear nuclei
(for review see Frisina and Walton, 2006). A decreased function of the
inhibitory system, mostly glycinergic, results in an increased ring of
neurons. A similarly decreased function of the GABAergic inhibitory
system at higher levels of the auditory pathway (for review see
Caspary et al., 2008) contributes to the complexity of the changes occurring in the auditory system during aging. All of these changes then inuence the situation in the auditory cortex, and it is difcult to separate
the effects of individual components from the overall nal picture.
5. Conclusions
In conclusion, the results of our study demonstrate that aging in man
is accompanied by signicant changes in the concentration of some
metabolites in the auditory cortex. Declining levels of the main excitatory transmitter glutamate may signicantly inuence the quality of
acoustical signal processing in the brain, particularly the most complex
acoustical signal speech. The declining levels of metabolites are
apparently not highly correlated with the sensory deafferentation that
accompanies human aging and that are manifested by a gradual loss
of hair cells, neurons and lateral wall cells in the inner ear, starting as
early as in adolescence. This conclusion is based on the fact that our
two groups of elderly subjects, MP with a mild age-related loss of sensory receptors in the inner ear (as demonstrated by audiograms) and EP
with profound presbycusis connected to a greater loss of hair cells
(and with large hearing threshold shifts), show almost no differences
in their metabolite levels. Also, the distribution of the white and gray
matter and the cerebrospinal uid does not show any essential differences between MP and EP groups. Only the large difference in lactate
levels between subjects with expressed presbycusis and control young
subjects points at more serious metabolic dysfunction in these subjects.
Conict of interest
The authors have no conicts of interests.
Acknowledgments
This research was supported by the grant GACR P304/10/1872 and
the Institutional Support 00023001IKEM of the Ministry of Health of
the Czech Republic.
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