Cellular Neuroscience207 Ian Parker细胞神经科学207伊恩派克.ppt

Cellular Neuroscience (207)Ian Parker Lecture # 2 - Ion channels: electrophysiology Single ion channels Single channel kinetics How big are single channel currents? Single channel current and conductance Because single channel current varies with membrane potential and ion gradient, a better measure is the conductance of the channel (g). This is a fixed characteristic (‘fingerprint’) of a given channel. g = i / (V-Veq) (v = membrane potential : Veq = reversal potential for current flow through channel) Unit of conductance is the Siemen (S : 1/Ohm) : single channel conductances are expressed in pS Range of channel conductances Recording the activity of single channels‘Patch-clamp’ technique : Neher Sakmann, 1976, (Nobel Prize1991) A commercial patch-clamp amplifier The ‘giga-seal’ Gigaseal recording configurations What can patch-clamp recordings tell us? Mean channel open time is a characteristic of any particular type of channel, but individual openings vary randomly * cytosol extracellular Molecular structure Physical structure Functional model Simplified model Cell membrane Transitions from open to shut are instantaneous Mean open time is a fixed characteristic of the channel Mean closed time shortens with increasing stimulus (e.g. depolarization or agonist concentration) Single channel current depends on channel conductance and electrochemical gradient for ion flow Channel opening does not require energy source (ATP), so channel continues to work in isolated membrane patch. Energy for ion flow (current) comes from electrochemical gradient across membrane Amps (log scale) 1 10-3 (mA) 10-6 (mA) 10-9 (nA) 10-12 (pA) 10-15 (fA) 100 W light bulb calculator action potential at node of Ranvier e.p.s.c. (current evoked by 1 vesicle of neurotransmitter) Single channel currents One ion per ms Limit of conventional Voltage clamp 100 50 -50 -100 Membrane potential (mV) -1 1 Single c