Molecular Mechanisms of Pediatric Kidney Disease Nader Gordjani MD PhD Professor of Pediatrics Universities of Freiburg and Frankfurt.
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Molecular Mechanisms of Pediatric Kidney Disease
Nader Gordjani MD PhDProfessor of Pediatrics
Universities of Freiburg and Frankfurt
Molecular origins of renal diseases
H2O
Hypophasphatemic rickets
Bartter-Syndrome
Hypokalemic alkalosis:
Gitelman-S.
Diabetes insipidus renalis
NEPHROTIC SYNDROME ?
CYSTÎC KIDNEYS ?
Lesions sites of the glomerulus
Filtration barrier
TH-Lypmphocytes
Capillaries
Mesangial cells
s. Biopsie
Deposits of antibodiesand complement
Podocyte
Autoantibodies
Nephrotic Syndrome
Nephritic Syndrome
kidney
80% Steroid-sensitive
Stable function
20% Steroid-resistant
deterioration
Nephrotic Syndrome in Children
What is the role of the podocyte in nephrotic syndrome ?
Chemokines
cytokines
complement
autoantibodies
Inflammatoryagents
What is the role of the podocyte in nephrotic syndrome and as target of immunmodulation?
Chemokines
cytokines
complement
autoantibodies
Inflammatoryagents
Imm
unos
uppr
essi
on
Ca2+- Signalling in the cell
Stimulation of:
Signalling pathwaysGene expressionChannel opening/closing …
0.0
1.0
2.0
3.0
fluo
resc
en
ce r
atio
34
0/3
80
ATP 10-4 M
C a2+ 10-6 M
3 m in
Calcium signalling in the Podocyte
• Fluorescence microscopy with Fura-2 AM Single-Photon TubeVideo-ImagingLaser-Scanning
RT-PCR• Isolation of rat glomeruli by sieving method• Statistical analysis: student´s t-test
chemokine receptor EC50 [ng/ml]
Rantes 1, 3, 5 50-700
TECK 4 300
Eotaxin 3 50
MIP-1 1, 5 20
MIP-1 5 100
MCP-1 2 10
MCP-2 2 200
MCP-3 1, 2 200
TARC 4 10
CXC
IL-8 1, 2 200
0.0
0.5
1.0
1.5
2.0flu
ore
scen
ce r
atio
34
0/3
80
ATP 100 µm ol/l
R AN TES 0.4 µg/m l
M C P-1 0.2 µg/m l
M IP-1 60 ng/m l
5 m in
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
f
luor
esce
nce
rat
io 3
40/3
80 n
m
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0.8
1.0
1.2
1.4
f
luor
esc
ence
rat
io 3
40/
380
nm
(n=5) (n=8)
CyA
ATP A II
CyA
Freshly isolated rat glomerulum
0
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1
1.5
2
2.5
3
3.5
4flu
ores
cenc
e ra
tio 3
45/3
80 n
m
AC H 1 µm ol/l
M C P-1 20 ng/m l
10 m in
C YA 10 nm ol/l
R AN TES 70 ng/m l
ConclusionsChemokines induce an increase of [Ca2+]i in mouse podocytes by release from cytosolic Ca2+-stores and thus stimulate Ca2+-mediated signal transduction.
Therefore podocytes are target cells of these proinflammatory factors.
ATP also causes a characteristic rise of [Ca2+]i, which is mediated by Ca2+-influx from the extracellular space in addition to store release.
Cyclosporine A did not influence the chemokine- or ATP-associated Ca2+-effects.
These findings could partly be confirmed in podocytes from freshly isolated intact rat glomeruli.
Physiology
Andreas BenesicRuth FreudingerMichael GekleGerald Schwerdt
Pediatrics
Nader GordjaniAntje KirchhoffBrigitte Wollny
University of Würzburg
University of Freiburg
Physiologie
Rainer GregerHermann PavenstädtJens LeipzigerRoland NitschkeViktoria Munzinger
Hildebrandt F et al. JASN 2009;20:23-35
Ciliar defects: a unifying theory of cystic kidney disease?
Chapin et al. JCB 2010
Ciliar defects: a unifying theory of cystic kidney disease?
Hildebrandt F et al. JASN 2009;20:23-35
Correct mitotic spindle
orientation
False mitotic spindle
orientation
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