Author/Authors :
Meamar ، Rokhsareh نويسنده Department of Cell and Molecular Biology, Cell Science Research Center, Royan Institute for Animal Biotechnology, ACECR, Isfahan, Iran , , Karamali، Fereshte نويسنده Department of Cell and Molecular Biology, Cell Science Research Center, Royan Institute for Animal Biotechnology, ACECR, Isfahan, Iran , , Ali Mousavi، Seyed نويسنده Isfahan Neurosciences research center, Isfahan University of Medical Sciences, Isfahan, Iran , , Baharvand، Hossein نويسنده , , NASR ESFAHANI، MOHAMMAD HOSSEIN نويسنده ,
Abstract :
Objective: Ecstasy, or 3, 4 (±) methylenedioxymethamphetamine (MDMA), is a potent neurotoxic drug. One of the mechanisms for its toxicity is the secondary release of glutamate.
Mouse embryonic stem
cells (mESCs) express only one glutamate receptor,
the metabotropic
glutamate receptor 5
(mGlu5), which is involved in the maintenance and
self-renewal
of mESCs. This study aims to investigate whether MDMA could influence self-renewal via the mGlu5 receptor in mESCs.
Materials and Methods: In this expremental study, we used immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR) to determine the presence
of
the
mGlu5
receptor in mESCs.
The
expression
of
mGlu5
was
evaluated
after
MDMA
was added to mESCs throughout neural precursor cell formation as group 1
and
during neural precursor cell differentiation
as group 2. The
stemness characteristic
in
treated
mESCs
by
immunofluorescence
and
flow
cytometry
was
studied.
Finally,
caspase
activity was evaluated by fluorescence staining in the treated group. Oneway
ANOVA
or repeated measure of ANOVA
according to the experimental design
was
used for statistical analyses.
Results: In this study mGlu5 expression was shown in mESCs. In terms of neuronal differentiation, MDMA affected mGlu5 expression during neural precursor cell formation
(group
1)
and
not
during
neural
precursor
differentiation
(group
2).
MDMA
(450
µM)
induced a significant increment in self-renewal properties in mESCs but did not
reverse
2-methyl-6(phenylethynyl) pyridine (MPEP,
1 µM), a non-competitive selective
mGlu5
antagonist.
Fluorescence
staining
with
anti-caspase
3
showed
a
significant
increase in the number of apoptotic cells in the MDMA
group.
Conclusion: We observed a dual role for MDMA on mESCs: reduced proliferation and maintenance of self-renewal. The lack of decreasing stemness characteristic in presence
of
MPEP
suggests
that
MDMA
mediates
its
role
through
a
different
mechanism
that
requires further investigation. In conclusion, despite being toxic, MDMA
maintains
stemness
characteristics.
