Mol Neurobiol (2013) 48:921–930
DOI 10.1007/s12035-013-8480-0
The Imbalanced Expression of Adenosine Receptors
in an Epilepsy Model Corrected Using Targeted
Mesenchymal Stem Cell Transplantation
Kang Huicong & Xue Zheng & Wang Furong & Tang Zhouping &
Xu Feng & Hu Qi & Liu Xiaoyan & Huang Xiaojiang & Zhang Na &
Xu Ke & Zeng Zheng & Zhu Suiqiang
Received: 14 April 2013 / Accepted: 30 May 2013 / Published online: 20 June 2013
# Springer Science+Business Media New York 2013
Abstract Adenosine inhibits epileptic episodes by interacting
with G-protein-coupled receptors. This study examined the
mechanism by which the inhibitory effect of adenosine be-
comes impaired during epileptogenesis. Dynamic changes in
adenosine A1 receptors (A1Rs) and A2a receptors (A2aRs)
were investigated in a kindling model of epilepsy. RT-PCR,
Western blotting, and immunofluorescence results indicated
that expression of A1Rs was increased in the hippocampus
24 h after kindling, but progressively decreased 1 and 6 months
after kindling. Opposite changes were seen in the expression
of A2aRs. This bidirectional change resulted in an imbalance
between A1Rs and A2aRs and dysregulation of the adenosine
system. Autologous mesenchymal stem cell (MSC) transplan-
tation was used to correct this disorder and avoid side effects
of systematic adenosine therapy. Paramagnetic iron oxide
particles were used to mark and track the MSCs in vivo using
MRI. The results indicated that the transplanted cells migrated
along the callosum and settled at the ependymal layer. The
MSCs displayed a relatively long survival time, at least
3 months. The improved AR expression and EEG findings
suggested that MSC transplantation was a potentially ef-
fective means of treating refractory epilepsy.
Electronic supplementary material The online version of this article
(doi:10.1007/s12035-013-8480-0) contains supplementary material,
which is available to authorized users.
K. Huicong : X. Zheng : W. Furong : T. Zhouping : X. Feng :
H. Qi : L. Xiaoyan : H. Xiaojiang : Z. Na : X. Ke : Z. Zheng :
Z. Suiqiang (*)
Department of Neurology, Tongji Hospital Affiliated to Tongji
Medical College, Huazhong University of Science and Technology,
Jiefang Blvd., No.1095, Wuhan 430030, China
e-mail: zhusuiqiang@163.com
Keywords Adenosine receptors . Adenosine kinase .
Mesenchymal stem cells . Epilepsy . Transplantation
Introduction
The adenosine system plays an important role in neural
inhibition and has been shown to have anti-convulsant ac-
tivity in several epilepsy models [1–3]. The anti-epileptic
effects of adenosine are mainly mediated through two kinds
of high-affinity receptors: adenosine A1 receptors (A1Rs)
and A2a receptors (A2aRs) A2aRs [4]. Activation of A1Rs
inhibits voltage-gated Ca2+ channels and K+ channels, and
thereby blocks the release of glutamate and prevents neuro-
nal depolarization [5]. A2aRs are predominantly expressed
in the thalamus where it facilitates excitatory synaptic trans-
mission by counteracting A1R-mediated inhibition of gluta-
mate release [6]. The adenosine system is therefore regarded
as an “endogenous anti-epileptic system” [1]. Since seizures
and ARs are intimately related, researchers have tried to
reduce seizures by activating or blocking the adenosine
system [7, 8]. However, if the AR system is abnormal, drugs
that act on ARs will be unable to reduce seizures. Affected
patients may, therefore, continue to suffer with refractory
epilepsy or even develop status epilepticus. Dynamic obser-
vation of changes in ARs may therefore assist in understand-
ing the mechanism of refractory epilepsy. In the first part of
this study, we dynamically examined the change of ARs in a
kindling model of chronic epilepsy.
Stem cell transplantation is emerging as a new therapeutic
approach for treating epilepsy and is the subject of a number
of ongoing studies. It is thought that transplanted stem cells
922
survive, migrate, and differentiate into the functional neural
cells, and thus correct AR abnormalities. Mesenchymal stem
cells (MSCs) have shown particular promise as autologous
implants. In this study, MSCs were transplanted into rats
with experimentally induced chronic epilepsy and their ef-
fect on AR expression and abnormal discharges were ob-
served. Our objective was to provide an experimental basis
for the application of stem cell transplantation in the treat-
ment of chronic epilepsy.
Materials and Methods
Animals
Specific pathogen-free male Wistar rats (weighing 220 to
250 g) were obtained from the Animal Center of Tongji
Medical College, Huazhong University of Science and
Technology, Wuhan, China. The animals were kept under
controlled conditions at 22±2 °C and 50–60 % humidity, in a
12-h dark/light cycle. The animals had free access to tap
water and standard feed. The rats were divided into three
groups of eight, comprising a kindled group, control group,
and normal group. Rats in the kindling group received intra-
peritoneal injection of lithium chloride–pilocarpine, those in
the control received normal saline and animals in the normal
group received no treatment.
The study protocol was approved by the Animal Care and
Use Committee of Hubei Province, China (Y20081342).
Throughout the study, international standards of animal
welfare were strictly followed in terms of care and use of
animals.
Establishment of the Epilepsy Model
Kindling was induced in accordance with previously de-
scribed methods [9]. In brief, animals were administered
intraperitoneal pilocarpine (50 mg/kg) 18 to 24 h after the
intraperitoneal injection of lithium chloride (3 mmol/kg).
Following this procedure, most animals developed limbic
status epilepticus, which was terminated by administration
of diazepam (10 mg/kg, i.p.) 40 min after onset.
The seizures were recorded on a five-point Racine Scale.
Grades IV and V represented full kindling. After a seizure-free
(latent) period, animals developed spontaneous recurrent seizures
during the subsequent chronic period [10]. The animals were
video-taped to observe these seizures.
RT-PCR Analysis
Rats were decapitated 24 h, 1 month, and 6 months after the
induction of epilepsy. The hippocampus was isolated and
harvested on ice for RNA extraction, using TRIzol reagent
Mol Neurobiol (2013) 48:921–930
(Invitrogen Co., Carlsbad, CA, USA). Complementary DNA
was synthesized from total RNA using a reverse transcrip-
tion kit (Fermentas Co.) according to the manufacturer's
instructions.
mRNA was detected by a reverse transcription-polymerase
primer chain reaction (RT-PCR) performed on a PTC-
100 programmable thermal controller (MJ Research, Inc.,
Watertown, MA, USA). All primers were designed using pub-
lished sequences and primer design software package (Primer
3). The following sequences were used for A1R (630 bp): 5′-
ATCCCACTGGCCA TCCTTATG-3′ and 5′-AGGCGATGTA
GATCAGAATGC-3′. The sequences for A2aR (540 bp) were:
5′-TGTCCTGGTCCTCACGCAGAG-3′ and 5′- CGGATCC
TGTAGGCGTAGA TGAAGG-3′. The thermal cycle profile
was as follows: 95 °C for 5 min, 95 °C for 30 s, 58 °C for 45 s,
72 °C for 1 min, repeated for 38 cycles, followed by 72 °C for
10 min to terminate the reaction. PCR products were separated
on 1.5 % agarose gels and visualized with ethidium bromide.
The density of each band was determined by using Image Quart
Software. Expression levels were indicated by the density
relative to that of β-actin which was an internal control.
Western Blotting
Protein was extracted from the tissue using lysate. After dilu-
tion in SDS-PAGE sample buffer, protein and pre-stained
molecular weight markers were separated by SDS-PAGE at
voltages reducing from 80 to 120 V. The products were electro-
transferred to a polyvinylidene difluoride membrane using a
current of 200 mA for 60 min. The blots were washed and
incubated in TBS-T+5 % non-fat dried milk for 60 min. They
were then re-incubated with primary antibodies (1:1,000 rab-
bit purified IgG anti adenosine; A1R, Abcam Co. and 1:100
mouse purified IgG anti adenosine A2aR, Upstate Co).
Thereafter, the blots were incubated overnight washed and
incubated for 1 h with secondary antibody labeled with horse-
radish peroxidase (Santa Cruz Biotechnology, Santacruz, CA,
USA). After three 10-min washes with Tris-buffered saline
and Tween the membrane was stained using a DAB kit
(Zhongshan Co., Beijing, China). Image density was ana-
lyzed using Image Quart Software. For each sample, the
optical density was normalized using β-actin as an internal
control.
Immunofluorescence
Frozen sections (10-mm thick) were prepared to detect
AR expression by immunofluorescence. Antibodies of
A1R and A2aR were diluted 1:100 and those of Cy3
and FITC were diluted 1:300. Positively stained cells
were identified by green or red membrane fluorescence.
Two slices were selected from each animal and 10 fields
were observed at a high magnification.
Mol Neurobiol (2013) 48:921–930 923

Fig. 1 Expression of A1Rs and

A2aRs in the hippocampus after
kindling detected by RT-PCR
and Western blotting. The upper
panels show the RT-PCR
electrophoretogram for A1Rs
and A2aRs. A1R expression
was up-regulated at 24 h and
down-regulated progressively at
1 and 6 months after kindling.
Changes in A2aRs were in the
opposite direction. The lower
panel shows Western blotting
results. The dynamic changes in
A1R and A2aR protein were
consistent with RT-PCR
findings

Culture, Labeling and Identification of MSCs
Wistar rats 4 to 5 weeks old were sacrificed by decapitation
and dipped into 75 % alcohol for 8 min. Whole tibial and
fibular bones were aseptically isolated and separated from
muscle and bone membranes in iced phosphate buffer solu-
tion (PBS). The epiphyses were removed and the marrow
cavity was washed three times in PBS. The flushing fluid
was centrifuged at 1000 r/min for 10 min. The resulting cells
were suspended in a complete medium (containing 80 %
DMEM low glucose medium, 18 % fetal calf serum, 1 %
P-S, 1 % glutamate, EGF) and cultured at 37 °C and in 5 %
CO2 and 95 % humidity.
MSCs were labeled with paramagnetic iron oxide parti-
cles (PIOPs) provided by Prof. Zhu Wenzhen (Department of
Radiology, Tongji Hospital). PIOPs (50 μg Fe/mL) were
added to the complete medium. Poly-L-lysine (PLL; Sigma,
USA) was added at a ratio of Fe/PLL of 1:0.03, and the
samples were incubated at 37°C for 1 h. Half of the MSC
medium (1.5 mL containing fifth-generation MSCs) was
replaced medium containing PIOP and the resultant samples
were incubated at 37 °C for 24 h. Complete medium was

Table 1 Quantitative RT-PCR data for hippocampal AR expression (A/Aβ-actin)

Group N A1R A2aR

24 h 1 months 6 months F value 24 h 1 months 6 months F value

KG 8 1.1483±0.1182 0.7749±0.0262 0.5682±0.0443 311.987 0.8338±0.0572 1.1581±0.0447 1.2169±0.0332 289.735

CG 8 0.8997±0.0170 0.8846±0.0354 0.8943±0.0281 0.221 1.0016±0.0045 0.9983±0.0165 0.9760±0.0229 0.519
NG 8 0.9973±0.0194 1.0061±0.0372 1.0175±0.0143 0.953 0.9842±0.0782 1.0037±0.0358 0.9953±0.0147 0.348

Data are shown as mean±SD

N number of animals in each group, KG kindling group, CG control group, NG normal group
924 Mol Neurobiol (2013) 48:921–930

Table 2 Quantitative Western Blot data for hippocampal ARs expression (A/Aβ-actin)

Group N A1R A2aR

24 h 1 month 6 months F value 24 h 1 month 6 months F value

KG 8 0.7872±0.0621 0.4532±0.0380 0.2633±0.0609 269.782 0.2098±0.0257 0.5108±0.03210 0.7080±0.0371 329.691

CG 8 0.5829±0.0237 0.5417±0.0573 0.5782±0.018 2.643 0.3428±0.0304 0.3643±0.05190 3316±0.0413 1.927
NG 8 0.6271±0.0592 0.5913±0.0620 0.5748±0.047 2.041 0.2975±0.0469 0.3214±0.03520 3195±0.0583 1.538

Data are shown as mean±SD

N number of animals in each group, KG kindling group, CG control group, NG normal group

then added and the MSCs were incubated at 37 °C and in 5 %
CO2 for 2 days.
Fourth- and fifth-generation MSCs were washed with PBS
containing 1 % bovine serum albumin and incubated with
CD45, CD90, CD105 antibodies for 15 min at 4 °C. After
incubation, the cells were washed twice with PBS and analyzed
on a FACSAria system with built-in FACS DIVA 6.2 Software
(BD Immunocytometry Systems, Heidelberg, Germany).
Six-well plates were cover-slipped and coated with PLL.
Then, cells were digested using 0.5 % trypsin and the
suspended cells were seeded into six-well plates. Cells were
fixed with 4 % paraformaldehyde for 20 min, washed three
times, and incubated with 2 % potassium ferrocyanide (Perl’s
reagent) in 6 % hydrochloric acid for 30 min. After another
wash, the cells were counterstained with 0.5 % eosin for
3 min. Cells that had not been subjected to PIOPs labeling
were used as controls.
MSCs Transplantation, MRI Tracking and In Vivo
Histological Identification
Cells were harvested and re-suspended at a concentration of
2×106/mL in PBS. One month after kindling, the rats were
anesthetized by intraperitoneal injection of 6 % chloral hy-
drate (at 0.5 mL/100 g). The rats were placed in a stereotaxic
instrument (SR-5, Narishige, Japan). The right hippocampus
was located (3.8 mm behind the frontal fontanelle, 3.2 mm
from the midline, and 3.5 mm deep) and the labeled MSCs
were transplanted using a microinjector. A total of 2 μL of
cell suspension was injected at a rate of 0.2 μL/min. After
injection, the microinjector remain in situ for 10 to 15 min to
avoid cell reflux.
Seven days, 1, and 3 month(s) after transplantation, the
labeled MSCs were tracked in vivo by 1.5 T MRI. The rats
were anesthetized and placed into a specially designed receiv-
er MR-coil. Fast spin echo T2-weighted images were obtained
using the following parameters: repetition time/echo time
4,400/80 ms, echo train 6, slice thickness 1.5 mm, matrix
64×64, and four excitations.
Three months after transplantation, frozen sections (10 mm
thick) were made for Prussian blue staining using previously
reported methods.
EEG Detection
EEG recording was carried out on day 15 after kindling and
was repeated 3 months after transplantation. Two electrodes
were separately placed on the frontal lobe (2.0 mm in front
of the anterior fontanelle, 2.0 mm away from the midline
and 0.5 mm deep) and hippocampus (2.0 mm behind the
anterior fontanelle, 3.2 mm away from the midline and
3.5 mm deep). The other electrodes served as a ground-
wire and reference electrode respectively. The recording
trace was set at a speed of 15 mm/s with 2 mm displacement
representing 50 V. Each recording lasted 15 min. The aver-
age frequency of spike waves (total number of the spike
waves divided by recording time) was calculated and am-
plitude was measured.

Table 3 Positive fluorescence cell count for hippocampal ARs (A/Aβ-actin)

Group N A1R A2aR

24 h 1 month 6 months F value 24 h 1 month 6 months F value

KG 8 76.17±4.62 49.83±4.36 38.50±4.81 166.619 43.83±5.12 90.33±5.43 114.50±4.04 179.852

CG 8 62.67±5.57 64.39±3.78 65.81±4.39 1.674 1.674 64.17±4.17 66.82±4.29 64.98±5.61 1.42
NG 8 59.73±4.16 57.20±5.24 61.53±3.92 2.68 66.72±3.26 63.23±5.08 65.37±4.90 1.97

Data are shown as mean±SD

N number of animals in each group, KG kindling group, CG control group, NG normal group
Mol Neurobiol (2013) 48:921–930 925

Fig. 2 Immunofluorescence of
hippocampal ARs in after
kindling. a A1R expression in
normal group. b A1R
expression 24 h after kindling. c
A1R expression 1 month after
kindling. d A1R expression
6 months after kindling. e A2aR
expression in normal group. f
A2aR expression 24 h after
kindling. g A2aR expression
1 month after kindling. h A2aR
expression at 6 months after
kindling
926
Statistical Analysis
The data were analyzed using the SPSS 12.0 Software.
Numerical data were expressed as means and standard de-
viations (±SD). Between group differences were evaluated
using one-way ANOVA analysis. Values of P<0.05 were
considered statistically significant.
Results
Bidirectional Change of ARs in Hippocampus
After Kindling
The expression of A1Rs in the hippocampus increased tran-
siently 24 h after kindling, and thereafter decreased progres-
sively 1 and 6 months after kindling. A2aR expression
decreased 24 h after kindling and then increased at 1 and
6 months Adenosine receptor expression remained
unchanged in the hippocampus in the control and normal
group. The results are shown in full in Fig. 1 and Table 1.
There were statistically significant differences in post-
kindling A2aR and A1R expression between kindling and
control groups (P<0.05 or P<0.01). There was also a statis-
tically significant difference in AR expression between the 1
and 6 month assessments (P<0.0001).
Western blotting and immunofluorescence detection at
different time points also exhibited significant bidirectional
changes ARs expression after kindling (Figs. 1 and 2 and
Tables 2 and 3).
MSC Identification and Labeled by PIOPs
Primary cultured MSCs which were spindle-shaped and
proliferated rapidly and became fibroblast-like in shape after
5 to 7 days. They were positive for CD90 and CD105, but
negative for CD45 (S1).
Prussian blue staining revealed the presence of blue par-
ticles in the cytoplasm of virtually all MSCs that had been
cultured in the presence of PIOPs. PIOP-labeled cells were
all positively stained and non-labeled cells (control cells)
were all negatively stained. The PIOPs were transferrable
to daughter cells such that positive PIOP-labeling in the fifth
generation was estimated at 87.4 % (Fig. 3).
Post Transplantation Survival and Immigration of MSCs
PIOP-labeled MSCs showed low signals on T2-weighted
images at all time points after transplantation and scan indi-
cated that the cells migrated towards the midline of brain
(Fig. 3). Prussian blue staining showed that the transplanted
cells had migrated to the callosum and were distributed along
the endyma at 3 months post transplantation (Fig. 3).
Mol Neurobiol (2013) 48:921–930
Fig. 3 Prussian blue MSC staining and tracking by MRI and histology.„
The upper panels show Prussian blue staining with numerous blue-
stained iron particles in the cytoplasm of all cells (right). Notably,
87.4 % of cells remained labeled after five culture transfers (middle).
The left panel shows the absence of staining in control cells. The middle
panels show T2-weighted MRI images displaying dark cell clusters at
7 days (left), 1 month (middle) and 3 months (right) after labeled MSCs
transplantation. Labeled cells migrated towards the midline of brain.
The lower panels show histology of brain tissue slices after PIOP-
labeled MSCs transplantation. a Transplanted cells migrated towards
the callosum and round cell clusters became fusiform. b A blue-stained
cell zone was distributed along the endyma
Improved A1Rs/A2aRs Ratio After MSCs Transplantation
Three months post transplantation, A1Rs expression in the
kindling group was up-regulated and A2aR expression was
down-regulated relative to the control group. This resulted in
an improved A1R/A2aR ratio in the kindling group (Fig. 4).
Post Transplantation Epileptic-Form Discharge
The pre transplantation frequency of sharp waves was
19.65±2.18 times per sheet and the amplitude was
297.43±25.37 mV. Post transplantation, the frequency de-
creased to 7.48±0.91 times per sheet and the amplitude to
86.27±7.72 mV (both P<0.0001; Fig. 4). No statistically
significant changes were seen in the control group.
Discussion
A reduced density of A1Rs has been reported in patients with
chronic epilepsy [11] and in animal models [12]. By contrast,
expression of A2aRs has been found to increased sharply in
patients with epilepsy [13]. In this study, we examined dy-
namic changes in ARs expression in a rat kindling model,
from the time of acute insult until the development of chronic
epileptogenesis. Our finding that A1R expression was signif-
icantly up-regulated 24 h after kindling was consistent with
previous studies reporting upregulation of AIRs between 1
and 24 h after kindling with pentetrazole [14, 15]. Some
researchers believe that this is an adaptive reaction to acute
seizures [16]. Other studies have concluded that the decreased
A1R density reflected the degeneration of neural cells [12].
Based on these observations, it has been proposed that an
acute increase in extracellular adenosine level and expression
of A1Rs may enhance the inhibitory action of adenosine
system until the point where the seizure is terminated.
In the present study, we found that A1R expression was
progressively reduced 1 and 6 months after the experimental
onset of seizure. Other workers have described this progres-
sively reduced sensitivity of A1Rs to extracellular adenosine
in terms of a “desensitization” response [17]. This reduction in
A1R-mediated inhibition may be an important mechanism
Mol Neurobiol (2013) 48:921–930 927

responsible for refractory epilepsy. There is evidence to indicate nerve fibers and alter neuronal plasticity. The reduced inhibito-
that adenosine, ATP, and activation of A1Rs inhibit regrowth of ry action over time may, therefore, promote neuronal regrowth
928 Mol Neurobiol (2013) 48:921–930

Fig. 4 Effect of MSCs

transplantation on the expression
of ARs and EEG recording. a–b
A1R expression in the
hippocampus. c–d A1R
expression in the temporal lobe.
e–f A2aR expression in the
thalamencephalon (left sham,
right transplantation group).
Middle panel/Bar chart *P<0.05
and Δ P<0.01compared with
control group. Lower panel EEG
recordings indicated that the
amplitude and frequency of
spike waves was reduced after
transplantation (150 mV/cm).
Cor temporal cortical electrode,
Hip hippocampal electrode
Mol Neurobiol (2013) 48:921–930
resulting in overactivity of the neural loop until neural plastic-
ity is changed [18]. Our results demonstrate a bidirectional
change in A1R expression as seizures progressed from the
acute to the chronic phase, suggesting that the adenosine
system failed to maintain an adaptive response to the seizures,
ultimately resulting in pathological damage. We also showed
that A2aR expression was down-regulated during the acute
phase of epilepsy, possibly reflecting the known antagonistic
action of A2aRs against A1Rs [19]. Thus, downregulation of
A2aRs and upregulation of A1Rs appear to work in tandem to
rapidly terminate seizures.
In the healthy brain, the ratio of A1Rs and A2aRs is in
balance to maintain the inhibitory action of adenosine sys-
tem. However, in chronic epilepsy, A1R expression in brain
tissue appears to be reduced and A2aRs appears to be in-
creased, which results in a weaker inhibitory effect of A1Rs.
Targeting ARs may, therefore, provide a new approach for
the treatment of epilepsy.
Cell transplantation techniques have been developed ex-
perimentally to avoid the peripheral side effects of systemic
adenosine, its antagonists, and agonists. It has been shown that
very low doses of adenosine (20 to 50 ng/day) substantially
inhibit epileptic-form discharge in experimental models [20].
The level of adenosine secreted by the transplanted MSCs is
low, and does not promote internal secretion of adenosine as
MSCs have no effect on adenosine transporters on the pre-
synaptic membrane [21]. Furthermore, recovery of the inhib-
itory effects of adenosine and its receptors post transplantation
is accompanied by increase adenosine levels and improve
adenosine receptor function.
MSCs have become a popular source of adult stem cells as
they are readily available, suitable for autologous transplan-
tation and can be induced to differentiate into neural cells.
Stable genetic modification of MSCs can also readily be
achieved [21].
Although the present did not examine adenosine secretion
by MSCs, previous workers have shown that MSCs produce
adenosine at a rate of 8.5 ng/h·105 [22]. In our study, the
transplanted MSCs remained alive 3 months post transplanta-
tion, indicating prolonged survival and proliferative capacity.
The marked increase in A1R expression and the marked
decrease in A2aR expression post transplantation suggested
that MSCs were able to reverse the imbalance of ARs that
characterizes epilepsy. Other researchers [23] reported that
transplantation of embryonic stem cells reduced seizure fre-
quency and epileptic-form discharges in epileptic animals,
suggesting that the transplanted cells were able to differentiate
into functional neurons that interact with other neural cells.
Long-term administration of adenosine and its analogues
has been shown to aggravate the dysfunctional adenosine
system and increase seizures [24]. However, this does not
appear to be an issue with adenosine derived from
transplanted stem cells. A previous study reported reduced
929
seizure occurrence for at least 8 weeks following stem cell
transplantation [25]. Some of the transplanted cells died over
this period of time and as a consequence, adenosine secretion
was gradually decreased and seizure episodes increased [25].
In our study, both the frequency and the amplitude of
epileptic-form discharges were reduced over a period of
3 months indicating that the anti-epileptic function can be
maintained for relatively long time post transplantation.
Other workers have shown that the density of ARs only
changed when they were stimulated by at least, millimole
levels of adenosine [26]. Their explanation was that adeno-
sine A3 receptors play a regulatory role in controlling the
adenosine system, and consequently the nanomole levels of
adenosine secreted by transplanted cells was not sufficient to
change AR density. Other workers have shown that low
levels of adenosine act as a tonic ARs inhibitory mechanism
[27] and thereby protect ARs from losing sensitivity and
density which is a potential advantage of cell transplantation
over other alternatives.
Conclusions
The present study was undertaken to attempt to restore the
normal function of adenosine system using MSCs transplan-
tation. We successfully reduced the amplitude and frequency
of EEG spike waves and improved the dysregulated ARs
expression that accompanied experimentally induced epilep-
sy in rats. Our results indicate that with further development,
MSCs transplantation might provide a promising new form
of therapy for refractory epilepsy.
Acknowledgments This work was supported by the National Natural
Science Foundation of China (81201006), Natural Science Foundation
of Hubei Province (2011CDB201) and Science and Technology Re-
search Plan Project of Wuhan City (201161038339-02).
Conflict of Interest The authors declare that there were no conflicts
of interest for any of the participating authors.
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