1. 1. Potassium channel Function: 1.shape action potentials and set the resting membrane potential 2. maintaining vascular tone 3. secretion of hormones 4. ion transport

2. Types Voltage-gated potassium channel Inwardly rectifying potassium channel resting potassium channels" or "leak channels Calcium-activated potassium channel

3. 6TM: The voltage- gated K+ channels Kv, KCNQ, Kca2+

4. A-type currrent M-current

5. 2-TM. The inward rectifier K+ KIR1.x, KIR2.x, KIR3.x, KIR4.x, KIR5.x, KIR6.x and KIR7.x

7. 4-TM. Sensitive to halothane,insensitive to TEA and 4-AP Regulate cell volume

8. 2. Cell recording solutions and Protocol for Ca2+ 、 Na+ 、 K+

9. (1) Calcium channels and Protocol

10. 激活曲线 Protocol

12. 失活曲线 稳态失活曲线

14. 失活后再激活曲线

16. （ 2 ） Sodium channel

17. Protocol

23. （ 3 ） Potassium channel

24. ① Inwardly rectifying potassium channel(K IR ) 激活方案

28. ② The delayed rectifier current (IK) largely responsible for repolarization and consists of two components namely IKs and IKr.

30. Voltage-Time dependent activation IKtail

31. Time dependent activation IKtail

34. 豚鼠心室肌细胞

35. ③ KATP

36. K ATP

37. βcell Smooth muscle cells

40. inside-out patch. The pipette potential is -100 mV Whole cell recording, With ATP and GDP in the pipette solution, Vh-60

42. inside-out patch

43. Inside out-recording(-100mv) Bath solution (for the intracellular side of the membrane) included (mM): KCl 120, KOH 20, MgCl2 1, EGTA 5, Hepes 10, glucose 5, Na2ATP 0.3, and MgADP 0.5 (pH=7.2); while pipette solution (for the extracellular side of the membrane) contained (mM): KCl 140, MgCl2 2, EGTA 2, glucose 10, and Hepes 10 (pH=7.4). Why Vh-60? Answer:minimize Kca

44. Whole cell-recording The bath solution contained (mM): NaCl 140, KCl 5.4, MgCl2 1.2, HEPES 10, EGTA 2, glucose 10 (pH adjusted to 7.4 with NaOH). The pipette solution comprised (mM): KCl 140, MgCl2 1, EGTA 10, HEPES 10, glucose 5, Na2ATP 0.3, and MgADP 0.5 (pH=7.2)

45. The membrane potential recording The bath solution contained (mM): NaCl 140, KCl 5.4, MgCl2 1.2, HEPES 10, EGTA 2, glucose 10 (pH adjusted to 7.4 with NaOH). The pipette solution comprised (mM): KCl 140, MgCl2 1, EGTA 10, HEPES 10, glucose 5, nystatin 250 μg/ml

46. ④ IK,M------(KCNQ2-KCNQ3) the M-current characterized as a low- threshold, voltage-gated K+ current that is active around the resting membrane-potential and constitutes an important regulator of neuronal excitability a voltage-and time-dependent, low-threshold, slowly activating current.

47. Traditional M-current voltage-clamp protocol: holding potential was -30 mV; step potentials were -40 to -80 mV Activation from a holding potential of -60 mV in 5 mV increments. I KM tail

48. 0.3 nA, 200-ms current-pulse injection M- current

50. Predominant K+ current in many mature neurons that is initially activated at the subthreshold range of membrane potential and deactivated during depolarizing pulses of duration. ⑤ Transient inactivating A-type current (IA)

51. 50ms

52. 14 (A1 ), 10 (A2 ), or 30 (A3 ) pA for 800 ms

53. ⑥ Kca

54. BK SK IK voltage gated and calcium modulated

55. NATURE 2004,427(26)

56. voltage-clamped at-30mV and pulsed for 400 ms from-100mV to +140mV For the perforated-patch experiments, a stock solution of 100 mg/ ml amphotericin B and final concentration of 200 mg/ml amphotericin The pipette solution contained (in mM): 140 KCl, 10 NaCl, 2 MgCl2, 0.7 CaCl2,1 EGTA and 10 HEPES (made to pH 7.3 with KOH). Cells were perfused with anextracellular buffer containing (in mM): 140 NaCl, 2.5 KCl, 0.5 MgCl2, 1.2 CaCl2,101 HEPES and 5 glucose (made to pH 7.4 with NaOH).

57. Journal of physiology a holding potential of –70 mV to test potentials between –50 and +80 mV in 10-mV increments.

58. To obtain steady-state KCa channel measurements in perforated patch-clamp experiments, Ca2+ sparks and thus transient KCa currents were abolished using thapsigargin (100 nM), an inhibitor of sarcoplasmic reticulum Ca2+- ATPase the bathing solution contained (in mM) 134 NaCl, 6 KCl, 2 CaCl2, 1 MgCl2, 10 HEPES, and 10 glucose (pH 7.4, NaOH). The perforated-patch pipette solution contained (in mM) 110 potassium aspartate, 30 KCl, 10 NaCl, 1 MgCl2, 10 HEPES, and 0.05 EGTA (pH 7.2 with KOH). Amphotericin B (1 μg/ml)

60. For inside-out recordings, the pipette solution contained (in mM) 140 NaCl, 6 KCl, 10 HEPES, 1 CaCl2, and 2 MgCl2. The inside-out bath solution contained (in mM) 140 KCl, 10 HEPES, 2 MgCl2, 5 or 1 EGTA, 1.6 HEDTA, and appropriate amounts of CaCl2 to obtain free Ca2+ concentrations of 0.1, 0.3, 1, 3, 10, 30, or 100 μM.