HAL Id: inserm-00654278
https://www.hal.inserm.fr/inserm-00654278
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Mineralocorticoid receptor overexpression facilitates
differentiation and promotes survival of embryonic stem
cell-derived neurons.
Mathilde Munier, Fredéric Law, Géri Meduri, Damien Le Menuet, Marc
Lombes
To cite this version:
Mathilde Munier, Fredéric Law, Géri Meduri, Damien Le Menuet, Marc Lombes. Mineralocorti-coid receptor overexpression facilitates differentiation and promotes survival of embryonic stem cell-derived neurons.. Endocrinology, Endocrine Society, 2012, 153 (3), pp.1330-40. �10.1210/en.2011-1436�. �inserm-00654278�
A
C
B
F
βtubulin III MR MergeWT WT P1-hMR P1-hMR ES Neurons 110 kDa MR GAPDH 36 kDa Rao 1 1.6 4.3 7.4 x 1.6 x 1.7 0 1 2 3 4 5 6 ES Neurons Re la v e mM R ex pr es si on mMR mRNA expression WT P1-hMR 0 10 20 30 40 50 60 70 ES Neurons Re la v e mG R ex pr es si on mGR mRNA expression WT P1-hMR
D
ns ns hMR protein expression 0 1 2 3 4 5 GR GAPDH WT P1-hMR 90 kDa 50 kDaNeuronal GR protein expression
E
ES Neurons Re la v e h M R ex pr es si on 0 0.2 0.4 0.6 0.8 1 WT P1-hMR WT P1-hMR Re la v e G R ex pr es si on nsC
B
0 10 20 ES Neurons Re la ve N es n ex pr es si onNes n expression
WT P1-hMRA
0 150 300 450 ES Neurons Re la ve M A P2 e xp re ss io nMAP2 expression
WT P1-hMR *** 50 kDa 36 kDa βtubulin III GAPDH WT WT P1-hMR P1-hMR ES Neurons Ra o 1 1.7 nsWT P1-hMR
Neurons
Caspase 3 35 kDa 17 kDaB
A
WTNeurons
P1-hMR 36 kDa PCNA GAPDH 36 kDa 0.0 0.7 1.4 WT P1-hMR Re la ve P CN A p ro te in le ve lPCNA protein level
WT P1-hMR Clea ved c aspase 3/ caspase 3 r atio ns WT P1-hMR WT P1-hMR Cleaved-Caspase 3 0 0,025 0,05 **
A
B
C
E
D
***
***
0 5 10 15 20 25 30 ES Neurons Re la ve B cl2 ex pr es si onBcl
2expression
WT P1-hMR***
0 1 2 3 ES Neurons Re la ve B clxL ex pr es si onBcl
xLexpression
WT P1-hMR***
0.0 0.2 0.4 0.6 0.8 1.0 1.2 ES Neurons Re la ve B ax e xp re ss io nBax expression
WT P1-hMR***
0.0 0.5 1.0 1.5 ES Neurons Re la ve B ak e xp re ss io nBak expression
WT P1-hMRRa o mRNA Bcl2/Bax Bcl2/Bak BclxL/Bax BclxL/Bak WT neurons 12.9 13.7 1.94 2.06 neuronsP1-hMR 23.4 19.2 2.54 2.08 WT P1-hMR Neurons Bcl2 GAPDH 28 kDa 36 kDa Neurons WT P1-hMR Bax GAPDH 20 kDa 36 kDa Bcl2/ Bax WT neurons 1.4 P1-hMR neurons 2.2
F
Protein ratioA
B
C
D
E
F
*** *** 0.0 0.6 1.2 scr MR si1 MR si2 MR Re la ve h M R ex pr es si onhMR expression
*** ** 0.0 0.6 1.2 scr MR si1 MR si2 MR Re la ve m M R ex pr es si onmMR expression
** *** 0.0 0.7 1.4 scr MR si1 MR si2 MR Re la ve M A P2 e xp re ss io nMAP2 expression
** ** 0.0 0.9 1.8 scr MR si1 MR si2 MR Re la ve B ax e xp re ss io nBax expression
** ** 0.0 0.6 1.2 scr MR si1 MR si2 MR *** 0.0 0.6 1.2 scr MR si1 MR si2 MR Re la ve B cl2 e xp re ss io nBcl2 expression
Bcl 2 /Ba x r atioBcl
2/Bax ratio
A
B
* ns ns 0 1 2 3C ALDO ALDO + CORT CORT +
** * ns ns 0.0 0.6 1.2 C ALDO ALDO +
RU486 CORT CORT + RU486
WT neurons
Bcl
2/Bax ratio
P1-hMR neurons
Bcl
2/Bax ratio
Bcl 2 /Ba x r atio Bcl 2 /Ba x r atioWT P1-hMR Bcl2 35 kDa 17 kDa 28 kDa 20 kDa
