Role of Solute Transport in

Cellular Organelles

Ke-Li Tsai, D.Phil.

Department of Physiology
Faculty of Medicine
Kaohsiung Medical University
Intracellular Ion Dynamics and Ca2+
Na2+
Homeostasis
Ca2+ Signalling and Transport in
Cellular Organelles
J. Cell Sci. 115: 3715–3716, 2002
Intracellular Ca2+ Homeostasis
[Ca 2+]o = 2×10-3M

IP3
Ca2+

[Ca 2+]I ≈ 10-8M

Germann & Stanfield (2004)
Calcium Response After Chemical
Stimulation
Hormone Receptor
Ca2+ ATPase Na+-Ca2+ exchanger Na+-K+ ATPase

ATP ATP
IP3

Ca2+ Ca2+

Ca2+
ATP Ca2+
sER Ca2+ ATPase
Capacitative Calcium Entry

 Stimulation of G-protein coupled receptor and

phospholipase C causes the release of Ca from
its internal store (mostly ER) via IP3 receptor Ca
release channel.

 After the store is depleted, there is a replenish of

calcium via plasma membrane Ca influx, which
is called capacitative Ca entry.
Capacitative Calcium Entry

Hormone Receptor
Na+-Ca2+ exchanger
Ca2+ ATPase

ATP
IP3

Ca2+
Capacitative Calcium Entry
Ca2+
ATP Ca2+
sER Ca2+ ATPase
Capacitative Calcium Entry and Store
Operated Calcium Channel

Contro (carbachol: cholinergic agonist)

+Gd3+
Endothelin-1 Effect on Ca2+ Signalling in
Astrocyte
 The level of endothelin-1 (ET-1) in brain tissue is
significantly increased after acute brain damage,
suggesting a role of endothelin-1 in astrocyte responses
after acute brain damage. J. Neurosci. Res. 47:590-602,
1997.

 Endothelin receptors are expressed by astrocytes and

regulate astrocyte hypertrophy in the normal and injured
CNS. Glia 41:180-190, 2003.

 ET-1 stimulated capacitative Ca2+ entry through ETA

receptors of a rat brain-derived type-1 astrocyte cell line.
Cell Signal. 15:197-207, 2003.
Capacitative Calcium Entry in Astrocyte
Capacitative Calcium Entry in Astrocyte

Tg: tharpsigargin
Capacitative Calcium Entry in Astrocyte
Capacitative Calcium Entry in Astrocyte

AA: arachidonic acid

Na+ and Ca2+ Signalling and
Transport in Cellular Organelles
 Cardiomyocyte
Adjacent
cardiomyocytes are
connected by
intercalated disks,
which have
desmosomes for
strength, and gap
junctions
for the coordination of
contraction during
pumping.

Vander et al. (2004)

Excitation-Contraction Coupling in Cardiac
Muscle

Germann & Stanfield (2005)

Na+ and Ca2+ Homeostasis in Cardiomyocytes

Cell Calcium 35: 603–612, 2004

 Ischemia–Reperfusion Injury
 Massive death of cardiomyocytes during
reperfusion after ischemia.

 Proposed mechanisms for reperfusion injury

 Free radical production
 Mitochondrial disruption
 “pH papradox”

NORMAL ISCHEMIA REPERFUSION

normal local cell death extensive cell death

Apoptosis Pathways in Cardiomyocytes

FASEB J. 16: 135–146, 2002

Cytochrome c Release Induces Apoptosis

Nature Cell Biol. 1: E209-E216, 1999

Electric Circuit of Mitochondrial Membrane
Potential Across Inner Membrane

m = -150 mV

Physiol. Rev. 80: 315-360, 2000

 Measurement of Mitochondrial Membrane
Potential with Fluoroprobe

TMRM+

Molecular Probes Lodish et al. (1999)

 Mitochondria-dependent Apoptosis

Cell death signal → MPTP

permeability ↑ →inner membrane
damage → ∆Ψm ↓ → cyt c release
→ apoptosis

Trends in Cell Biol. 10: 369-377, 2000

Mitochondrial membrane depolarisation during apoptosis recorded by TMRM+,
a potential probe
Am. J. Physiol. 282: F981-F990, 2002
Correlation between m Depolarisation and
Cytochrome c Release

Staurosporine (STS):
apoptosis-inducing drug

J. Biol. Chem. 274: 5654–5658, 1999

Mitochondria Ca2+ Overload Lead to Apoptosis

Nature Rev. Mol. Cell Biol. 4: 552-565, 2003

Hypothesis

Abnormal increase in mitochondrial Na+ causes

mitochondrial depolarisation and apoptosis in
cardiomyocytes
Intracellular Na+ Increase in H2O2-induced
Cardiomyocyte Apoptosis

[Na]i: intracellular Na
concentration

FASEB J. 18: 1442-1444, 2004

 Mitochondrial Na+ and Ca2+ Increase in
H2O2-induced Cardiomyocyte Apoptosis

[Ca]m
Rhod-2:
mitochondrial
Ca fluoroprobe

[Na]m
CoroNa Red:
mitochondrial
Na fluoroprobe

Mitochondrial Ca ([Ca]m) and Na ([Na]m) concentration measurement

Mitochondrial Na+ Increase in H2O2-induced
Cardiomyocyte Apoptosis
Mitochondrial Na+ concentration
([Na]m) measurement during
H2O2 treatment
Mitochondrial Na+ Increase in H2O2-induced
Cardiomyocyte Apoptosis

[Na]cyt: cytosolic Na
concentration
[Na]m: mitochondrial Na
concentration
Mitochondrial Na+ Involves in H2O2-induced
Cardiomyocyte Apoptosis

Mitochondrial Na+ concentration ([Na]m) is fixed at 65.9 mM by monesin

Mitochondrial Na+ Involves in H2O2-induced
Cardiomyocyte Apoptosis

La: mitochondrial Na/Ca

uniporter inhibitor

Mitochondrial Na+ concentration ([Na]m) is fixed at 65.9 mM by monesin

Mitochondrial Na+ Involves in H2O2-induced
Cardiomyocyte Apoptosis
Mitochondrial Na+ Involves in H2O2-induced
Cardiomyocyte Apoptosis
Sources of Materials
 Germann & Stanfield, Principles of Human Physiology, 2nd edn, Benjamin
Cummings, 2005
 Lodish et al., Molecular Cell Biology, 4th ed, W.H. Freeman, 1999
 Molecular Probes
(http://www.invitrogen.com/site/us/en/home/brands/Molecular-Probes.html)
 Vander, Sherman & Luciano, Human Physiology, 9th edn, Mc-Graw Hill,
2004