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Absence of S100A1 in mice is associated with a hypertensive phenotype dependent on both
gender and endothelial function
Jean-Francois Desjardins; Krystyna Kuliszewska; Adrian Quan; Duncan J. Stewart; Subodh Verma; Thomas G.
Parker
Presenter disclosure information
Jean-Francois Desjardins
Absence of S100A1 in mice is associated with a hypertensive phenotype dependent on both gender and endothelial function
Disclosures : none
Introduction
S100A1S100A1 Small dimeric EF-hand Ca2+-binding protein (20kDa)
Structurally related to Calmodulin Binds target proteins and regulate a variety of cellular processes
Phosphorylation and enzyme activity
Highly expressed in cardiomyocytes (0.2% total soluble protein content)
Positive regulator of cardiac performance (Most et al., (2001)
Proc Natl Acad Sci USA 98: 13889-94. )
Regulation of Ca2+ transient SERCA2 and RyR2
Introduction
S100A1S100A1 Expressed in endothelial and smooth
muscle cells (ECs and SMCs) ECsECs
Expression is increased in a rat model of cerebral basilar artery vasospasm (Lefranc et al., (2005) Neuropathol Appl Neurobiol 31(6): 649.)
SMCsSMCs Associates with the SR and actin stress fibers
(Mandinova et al., (1998) J Cell Sci 111: 2043.)
Objectives
To determine the impact of S100A1 deficiency on systemic BP
To assess the role of the endothelium in vascular function in S100A1 KO animals
To evaluate S100A1 and eNOS interaction
Methods
•S100A1 KO S100A1 KO **•WT (C57BL/6)WT (C57BL/6)
* Du et al., (2002) Mol Cell Biol 22(8):2821.
Human aortic Human aortic endothelial cellsendothelial cells
Ex vivoEx vivo vessel myograph isolated resistance and conduit vessels in KO and WT
In vivoIn vivo micromanometer catheter hemodynamics following coronary artery ligation or sham surgery impact of gender and anaesthesia was analysed
In vitroIn vitro eNOS/S100A1 co-immunoprecipitation and immunofluorescence co-localisation assays
S100A1 KO mice demonstrate poor survival and higher MAP following MI
0 10 20 300
50
100
WT
S100A1 KO
Time (days)
Su
rviv
al (
%)
n=49-56
P<0.01**
0
20
40
60
80
100
120
Sham MI
MA
P (
mm
Hg
)
WT
S100A1KO
#, P<0.05 vs.respective-WT (n=6-18)
#
#M
AP
(m
mH
g)M
AP
(m
mH
g)
ShamSham MIMI
0
20
40
60
80
100
120
Sham MI
MA
P (
mm
Hg
)
WT
S100A1KO
#, P<0.05 vs.respective-WT (n=6-18)
#
#M
AP
(m
mH
g)M
AP
(m
mH
g)
ShamSham MIMI
Male S100A1 KO exhibit higher mortality and MAP than female S100A1 KO mice
0 10 20 300
50
100
Female KO
Male KO
Time (days)
Su
rviv
al (
%)
P<0.05
n=20-36
**
0
20
40
60
80
100
120
140
Males Females
MA
P (
mm
Hg
)
WT
S100A1KO
* P<0.05 vs. female KO
# P<0.05 vs.respective-WT (n=6-18)
*
#
#
MalesMales FemalesFemales
MA
P (
mm
Hg)
MA
P (
mm
Hg)
0
20
40
60
80
100
120
140
Males Females
MA
P (
mm
Hg
)
WT
S100A1KO
0
20
40
60
80
100
120
140
Males Females
MA
P (
mm
Hg
)
WT
S100A1KO
* P<0.05 vs. female KO
# P<0.05 vs.respective-WT (n=6-18)
*
#
#
MalesMales FemalesFemales
MA
P (
mm
Hg)
MA
P (
mm
Hg)
The hypertensive phenotype is attenuated in S100A1 KO mice but persists under 2%
isoflurane anaesthesia
0
10
20
30
40
50
60
70
80
90
100
Males Females
MA
P (m
mH
g)
WT
S100A1KO
* P<0.05 vs. female KO
#, P<0.05 vs.respective-WT (n=8-20)
*#
#
0
10
20
30
40
50
60
70
80
90
100
Males Females
MA
P (m
mH
g)
WT
S100A1KO
* P<0.05 vs. female KO
#, P<0.05 vs.respective-WT (n=8-20)
*#
#
Male KO mice exhibit cardiac hypertrophy
3.0
4.0
5.0WT
S100A1 KO
MalesMales
HW
HW
// BW
(m
gB
W (
mg //
g)g) *
* P<0.05
n=18-23
3.0
4.0
5.0WT
S100A1 KO
3.0
4.0
5.0WT
S100A1 KO
3.0
4.0
5.0WT
S100A1 KO
3.0
4.0
5.0WT
S100A1 KO
MalesMales
HW
HW
// BW
(m
gB
W (
mg //
g)g) *
* P<0.05
n=18-23
* P<0.05
n=18-23
Male S100A1 KO mice exhibit non-characteristic increase in systolic pressure following IP injection
of the 2-adrenergic agonist xylazine
0 5 10 15 20 25
-25
0
25
50
WT n=5S100A1 KO n=4
Time (min)
L
VS
P (m
m H
g)
* P<0.01
*
** *
*
0 5 10 15 20 25
-25
0
25
50
WT n=5S100A1 KO n=4
Time (min)
L
VS
P (m
m H
g)
* P<0.01
*
** *
*
0 5 10 15 20 25
-200
-150
-100
-50
0
Time (min)
HR
(b
pm
)
Endothelium-dependent relaxation in isolated vessels is reduced in male
S100A1 KO
Mesenteric arteries
-8 -7 -6 -5
0
25
50
75
100
WT (n=5)S100A1 KO (n=5)
log Acetylcholine (M)
% R
ela
xa
tio
n
**PP<0.05<0.05
* **
**
Aortas
-8 -7 -6 -5
0
25
50
75
100
WT (n=5)S100A1 KO (n=4)
log Acetylcholine (M)
% R
ela
xa
tion
**PP<0.05<0.05
**
**
Mesenteric arteries
-8 -7 -6 -5
0
25
50
75
100
WT (n=5)S100A1 KO (n=5)
log Acetylcholine (M)
% R
ela
xa
tio
n
**PP<0.05<0.05
* **
**
Aortas
-8 -7 -6 -5
0
25
50
75
100
WT (n=5)S100A1 KO (n=4)
log Acetylcholine (M)
% R
ela
xa
tion
**PP<0.05<0.05
**
**
S100A1 KO and WT vessels exhibit similar vasoconstriction in response to
phenylephrine
Mesenteric arteriesMesenteric arteries AortasAortas
S100A1 and eNOS partly co-localize in cultured human aortic endothelial
cells (HAECs)
eNOSeNOS S100A1S100A1 mergedmerged
x60 x60 magmag
S100A1 and eNOS co-immunoprecipitates in cultured HAECs
IP: eNOSIP: eNOS
WB: eNOS, S100A1WB: eNOS, S100A1
eNOSeNOS
140 kDa140 kDa
S100A1S100A1
36 kDa36 kDaEGTA EGTA CCaa
2+2+
12hr12hr
Summary and conclusions• S100A1 plays a role in the regulation of basal blood
pressure in a gender specific manner
• Endothelium-dependent relaxation of isolated resistance and conduit vessels is reduced in male S100A1 KO animals
• eNOS and S100A1 co-immunoprecipitate in HAECs and partly co-localize around the nucleus
• S100A1 may be important not only in cardiac function in disease but also in the vascular responses