LIU Chuan Yong 刘传勇 Institute of Physiology Medical School of SDU Tel 88381175 (lab) 88382098 (office) Email: li

1. LIU Chuan Yong 刘传勇 Institute of Physiology Medical School of SDU Tel 88381175 (lab) 88382098 (office) Email: liucy@sdu.edu.cn Website: www.physiology.sdu.edu.cn

2. Section 2 Bioelectrical Phenomena of the Cell

3. Basic Concepts • Volt • A charge difference between two points in space

4. Basic Concepts • Ions – charged particles • Anions – Negatively charged particles • Cations – Positively charged particles

5. Basic ConceptsForces that determine ionic movement • Electrostatic forces • Opposite charges attract • Identical charges repel • Concentration forces • Diffusion – movement of ions through semipermeable membrane • Osmosis – movement of water from region of high concentration to low

6. Selective Permeability of Membranes • Some ions permitted to cross more easily than others • Neuronal membranes contain ion channels • Protein tubes that span the membrane • Some stay open all the time (nongated) • Some open on the occasion of an action potential, causing a change in the permeability of the membrane (gated)

7. I. Membrane Resting Potential • A constant potential difference across the resting cell membrane • Cell’s ability to fire an action potential is due to the cell’s ability to maintain the cellular resting potential at approximately –70 mV (-.07 volt) • The basic signaling properties of neurons are determined by changes in the resting potential

8. Membrane Resting Potential • Every neuron has a separation of electrical charge across its cell membrane. • The membrane potential results from a separation of positive and negative charges across the cell membrane.

9. Membrane Resting Potential • excess of positive charges outside and negative charges inside the membrane • maintained because the lipid bilayer acts as a barrier to the diffusion of ions • gives rise to an electrical potential difference, which ranges from about 60 to 70 mV. • (Microelectrode)

10. Concept of Resting Potential (RP) • A potential difference across the cell membrane at the rest stage or when the cell is not stimulated. • Property: • It is constant or stable • It is negative inside relative to the outside • Resting potentials are different in different cells.

11. Ion Channels • Two Types of Ion Channels • Gated • Non-Gated

12. Resting Membrane Potential • Na+ and Cl- are more concentrated outside the cell • K+ and organic anions (organic acids and proteins) are more concentrated inside.

13. Intracellular vs extracellular ion concentrations Ion Intracellular Extracellular Na+ 5-15 mM 145 mM K+ 140 mM 5 mM Mg2+ 0.5 mM 1-2 mM Ca2+ 10-7 mM 1-2 mM H+ 10-7.2 M (pH 7.2) 10-7.4 M (pH 7.4) Cl- 5-15 mM 110 mM

14. Resting Membrane Potential • Potassium ions, concentrated inside the cell tend to move outward down their concentration gradient through nongated potassium channels • But the relative excess of negative charge inside the membrane tend to push potassium ions out of the cell

15. Potassium equilibrium -90 mV

16. Resting Membrane Potential Na+ is more concentrated outside than inside and therefore tends to flow into the cell down its concentration gradient Na+ is driven into the cell by the electrical potential difference across the membrane. • But what about sodium? • Electrostatic and Chemical forces act together on Na+ ions to drive them into the cell • The Na+ channel close during the resting state

17. Na+ electrochemical gradient

18. Equilibrium Potentials • Theoretical voltage produced across the membrane if only 1 ion could diffuse through the membrane. • If membrane only permeable to K+, K+ diffuses until [K+] is at equilibrium. • Force of electrical attraction and diffusion are = opposite.

19. Calculating equilibrium potential Nernst Equation • Allows theoretical membrane potential to be calculated for particular ion. • Membrane potential that would exactly balance the diffusion gradient and prevent the net movement of a particular ion. • Value depends on the ratio of [ion] on the 2 sides of the membrane.

20. Nernst equation Equilibrium potential (mV) , Eion=ln [C]o RT zF [C]i where, [C]o and [C]i = extra and intracellular [ion] R = Universal gas constant (8.3 joules.K-1.mol-1) T = Absolute temperature (°K) F = Faraday constant (96,500 coulombs.mol-1) z = Charge of ion (Na+ = +1, Ca2+ = +2, Cl- = -1) For K+, with [K+]o = 4 mmol.l-1 and [K+]i = 150 mmol.l-1 At 37°C, EK= -97mV ENa = +60mv

21. [K+]o = 4 mmol.l-1

22. Resting Membrane Potential • Resting membrane potential is less than Ek because some Na+ can also enter the cell. • The slow rate of Na+ influx is accompanied by slow rate of K+ outflux. • Depends upon 2 factors: • Ratio of the concentrations of each ion on the 2 sides of the plasma membrane. • Specific permeability of membrane to each different ion. • Resting membrane potential of most cells ranges from - 65 to – 85 mV.

23. The Sodium-Potassium Pump extrudes Na+ from the cell while taking in K • Dissipation of ionic gradients is ultimately prevented by Na+-K+ pumps

24. Resting Potential

25. The formation of resting potential depends on: • Concentration difference of K+ across the membrane • Permeability of Na+ and K+ during the resting state • Na+-K+ pump

26. Factors that affect resting potential • Difference of K+ ion concentration across the membrane • Permeability of the membrane to Na+ and K+. • Action of Na+ pump

27. Basic Electrophysiological Terms I: • Polarization: a state in which membrane is polarized at rest, negative inside and positive outside. • Depolarization: the membrane potential becomes less negative than the resting potential (close to zero). • Hyperpolarization: the membrane potential is more negative than the resting level.

28. Basic Electrophysiological Terms I: • Reverspolarization: a reversal of membrane potential polarity. • The inside of a cell becomes positive relative to the outside. • Repolarization: restoration of normal polarization state of membrane. • a process in which the membrane potential returns toward from depolarized level to the normal resting membrane value.

29. II Action Potential • Successive Stages: • Resting Stage • Depolarization stage • Repolarization stage • After-potential stage (2) (3) (1) (4)

30. Concept • Action potential is a rapid, reversible, and conductive change of the membrane potential after the cell is stimulated. • Nerve signals are transmitted by action potentials.

31. Action Potential Sequence • Voltage-gated Na+ Channels open and Na+ rushes into the cell

32. Action Potential Sequence • At about +30 mV, Sodium channels close, but now, voltage-gated potassium channels open, causing an outflow of potassium, down its electrochemical gradient

33. Action Potential Sequence equilibrium potential of the cell is restored

34. Action Potential Sequence • The Sodium – Potassium Pump is left to clean up the mess…

35. Ion Permeability during the AP Figure 8-12: Refractory periods

36. Basic Electrophysiological Terms II (1) • Excitability: The ability of the cell to generate the action potential • Excitable cells: Cells that generate action potential during excitation. • in excitable cells (muscle, nerve, secretery cells), the action potential is the marker of excitation. • Some scholars even suggest that in excitable cells, action potential is identical to the excitation.

37. Basic Electrophysiological Terms II (2) • Stimulus: a sudden change of the (internal or external) environmental condition of the cell. • includes physical and chemical stimulus. • The electrical stimulus is often used for the physiological research. • Threshold (intensity): the lowest or minimal intensity of stimulus to elicit an action potential • (Three factors of the stimulation: intensity, duration, rate of intensity change)

38. Basic Electrophysiological Terms II (3) • Types of stimulus: • Threshold stimulus: The stimulus with the intensity equal to threshold • Subthreshold stimulus: The stimulus with the intensity weaker than the threshold • Suprathreshold stimulus: The stimulus with the intensity greater than the threshold.

39. Action Potential Summary • Reduction in membrane potential (depolarization) to "threshold" level leads to opening of Na+ channels, allowing Na+ to enter the cell • Interior becomes positive • The Na+ channels then close automatically followed by a period of inactivation. • K+ channels open, K+ leaves the cell and the interior again becomes negative. • Process lasts about 1/1000th of a second.

40. Properties of the Action Potential • “All or none” phenomenon • A threshold or suprathreshold stimulus applied to a single nerve fiber always initiate the same action potential with constant amplitude, time course and propagation velocity. • Propagation • Transmitted in both direction in a nerve fiber

41. III Initiation of Action Potential

42. Squid giant axon

43. Gated channel states

44. Na+ Channel a1-Subunit Structure NH2 Outside + + + + + + + + + + + + b1 Inside I I CO2H F F M M NH2 CO2H + + + + + + + + RVIRLARIGRILRLIKGAKGIR I II III IV IVS4 Voltage Sensor - Inactivation “Gate”

45. Voltage gated But “ready” Not “ready”

46. Activation & Fast Inactivation

47. Sodium Activation and Inactivation Variable vs Voltage Inactivation gate Activation Gate If resting potential depolarized by 15 – 20 mV, then activation gate opened with 5000x increase in Na+ permeability followed by inactivation gate close 1 ms later

48. Positive feedback loop graded Na+ potential If YES, then... Reach “threshold”? Stimulation V-gate Na+ channels open Na+ enters (depolarization)

49. Action potential initiation S.I.Z.

50. Action potential termination