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从小鼠背根神经节培养的施万细胞中的电压依赖性钙通道和钾通道。

Voltage-dependent calcium and potassium channels in Schwann cells cultured from dorsal root ganglia of the mouse.

作者信息

Amédée T, Ellie E, Dupouy B, Vincent J D

机构信息

Institut National de la Santé et de la Recherche Médicale, Unité de Neurobiologie Intégrative, Bordeaux, France.

出版信息

J Physiol. 1991 Sep;441:35-56. doi: 10.1113/jphysiol.1991.sp018737.

Abstract
  1. Whole-cell patch clamp studies were carried out on Schwann cells in organotypic cultures of dorsal root ganglia (DRG) from OF1 mice embryos (18-19 days). 2. In standard external solution, from a holding potential of -70 mV, two types of voltage-dependent K+ currents were recorded: a fast transient current and a delayed sustained current. With a holding potential of -30 mV, only the delayed sustained current could be evoked. 3. Both K+ currents were inhibited by tetraethylammonium chloride (TEA) and 4-aminopyridine (4-AP) in a dose-dependent manner. For the transient current the half-maximal effective dose was 100 mM for TEA and 1.3 mM for 4-AP. For the delayed sustained current the half-maximal effective dose was 11 mM for TEA and 4 mM for 4-AP. Both currents were insensitive to external Ca2+. 4. The delayed sustained current, isolated by use of a holding potential of -30 mV displayed a 'cumulative inactivation' which was removed by hyperpolarizing the membrane to -70 mV between each test pulse. 5. In K(+)-free external and pipette solutions, with 10 mM-external Ca2+, from a holding potential of -70 mV voltage-dependent Ca2+ channel currents were recorded. The threshold for activation was -45.3 +/- 5.4 mV (mean +/- S.D., n = 5) and the current inactivated fully at the end of the test potential. The current was unaffected by 2 microM-tetrodotoxin (TTX) and totally blocked by 5 mM-Co2+. 6. Equimolar replacement of external Ca2+ by Ba2+ did not significantly modify the voltage dependence (threshold for activation -42.8 +/- 6.4 mV, n = 7) or the magnitude of the inward current. Ca2+ and Ba2+ were equally permeant. The fully inactivating current was insensitive to both nifedipine and Bay K 8644 (1 microM each). Increasing the external Ba2+ concentration from 10 to 89 mM enhanced the Ba2+ current and shifted the voltage dependence of the current (threshold for activation, -30.5 +/- 7.3 mV, n = 9) along the voltage axis as expected for altered external surface potential. 7. In 89 mM-external Ba2+ solution, some cells displayed an additional slowly decaying current which was totally blocked by nifedipine (1 microM). 8. Ca2+ channel currents were recorded only when DRG neurons were present in the culture, as excision of explants and subsequent axonal degeneration led to loss of detectable Ca2+ channel currents. This phenomenon was never observed for K+ currents. 9. We conclude that mouse Schwann cells in organotypic culture possess voltage-dependent K+ and Ca2+ channels.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 对来自OF1小鼠胚胎(18 - 19天)背根神经节(DRG)的器官型培养物中的雪旺细胞进行了全细胞膜片钳研究。2. 在标准外部溶液中,从 - 70 mV的钳制电位记录到两种电压依赖性钾电流:一种快速瞬态电流和一种延迟持续电流。在 - 30 mV的钳制电位下,只能诱发延迟持续电流。3. 两种钾电流均被氯化四乙铵(TEA)和4 - 氨基吡啶(4 - AP)以剂量依赖性方式抑制。对于瞬态电流,TEA的半数最大有效剂量为100 mM,4 - AP为1.3 mM。对于延迟持续电流,TEA的半数最大有效剂量为11 mM,4 - AP为4 mM。两种电流对外部Ca2 +均不敏感。4. 通过使用 - 30 mV的钳制电位分离出的延迟持续电流表现出“累积失活”,在每个测试脉冲之间将膜超极化至 - 70 mV可消除这种失活。5. 在无钾的外部和移液管溶液中,加入10 mM外部Ca2 +,从 - 70 mV的钳制电位记录到电压依赖性Ca2 +通道电流。激活阈值为 - 45.3±5.4 mV(平均值±标准差,n = 5),电流在测试电位结束时完全失活。该电流不受2 μM河豚毒素(TTX)影响,被5 mM Co2 +完全阻断。6. 用Ba2 +等摩尔替代外部Ca2 +并未显著改变电压依赖性(激活阈值 - 42.8±6.4 mV,n = 7)或内向电流的大小。Ca2 +和Ba2 +具有同等通透性。完全失活的电流对硝苯地平和Bay K 8644(各1 μM)均不敏感。将外部Ba2 +浓度从10 mM增加到89 mM可增强Ba2 +电流,并使电流的电压依赖性(激活阈值, - 30.5±7.3 mV,n = 9)沿电压轴移动,这与外部表面电位改变时预期的情况一致。7. 在89 mM外部Ba2 +溶液中,一些细胞表现出额外的缓慢衰减电流,该电流被1 μM硝苯地平完全阻断。8. 仅当培养物中存在DRG神经元时才记录到Ca2 +通道电流,因为外植体切除和随后的轴突退变导致可检测到的Ca2 +通道电流丧失。这种现象在钾电流中从未观察到。9. 我们得出结论,器官型培养的小鼠雪旺细胞具有电压依赖性钾通道和钙通道。(摘要截断于400字)
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/616d/1180184/c690b162b3ce/jphysiol00441-0046-a.jpg

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