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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
