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Transcript
Excitable Cells: Electrophysiological and
Fluorimetrical Approaches
潘建源
神經細胞電生理實驗室
臺灣大學動物學研究所暨生命科學系
Calcium Signaling: Dynamics and Homeostasis
Stimulus-secretion
Coupling
Nature Reviews Molecular Cell Biology 4; 517-529 (2003)
4, 517 (2003)
Embryonic Cortical & Dopaminergic Neurons
E14.5 midbrain
50 m
TH
GFAP
TH: tyrosine hydroxylase, dopaminergic neuron marker
GFAP: glial fibrial associated protein, glia marker
Expressing GFP in cultured neurons
Bovine chromaffin cells
Adrenal
medulla
Adrenal
gland
Adrenal
cortex
STRESS
Nerve
signals
Hypothalamus
Releasing hormone
Kidney
Spinal cord
(cross
section)
Splanchnic nerve
Sympathetic
postganglionic
neurons
Nerve
cell
Anterior pituitary
Blood vessel
Nerve
cell
Adrenal
medulla
Epinephrine and
norepinephrine
SHORT-TERM STRESS RESPONSE
1. Glycogen broken down to glucose;
increased blood glucose
2. Increased blood pressure
3. Increased breathing rate
4. Increased metabolic rate
5. Change in blood-flow patterns, leading
to increased alertness and decreased
digestive and kidney activity
ACTH
Adrenal cortex
ACTH
Glucocorticoids
Mineralocorticoids
LONG-TERM STRESS RESPONSE
1. Retention of
sodium ions
and water by
kidneys
2. Increased
blood volume
and blood
pressure
1. Proteins and fats
broken down and
converted to
glucose, leading to
increased blood
glucose
2. Immune system
may be
suppressed
Biology, Campbell et al.
Electrophysiology and Electrochemistry
+20 mV
Whole-Cell Patch
Voltammetry
-80 mV
400 pA
Carbon
Fiber
e-
Oxidation
Oxidation Current
30 pA
ICa
INa
CA
[Ca2+]i
Capacitance
Trace
+
1s
High K+
150 ms
Action Potential
40 mV
0.2 s
Simultaneously monitor ionic current and [Ca2+]i fluorescence
response
[Ca2+]i
Exchanger Current
Double Patch to study synaptic connection
Synaptic connection
Post
EPSC
150 pA
50 m s
Pre
AP
30 m V
50 m s
Stimulus-secretion Coupling
1. What molecules are involved in: calcium binding
proteins, vesicle recycling-related proteins, Na+/Ca2+
exchanger, Ca2+ channels
2. How coupling is modulated: proteins, polyphenols,
lysophospholipids, toxins
3. Tracking vesicle movement