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大鼠CA1锥体神经元顶端树突中单个电压门控性Na⁺和Ca²⁺通道的特性研究

Characterization of single voltage-gated Na+ and Ca2+ channels in apical dendrites of rat CA1 pyramidal neurons.

作者信息

Magee J C, Johnston D

机构信息

Division of Neuroscience, Baylor College of Medicine, Houston, TX 77030, USA.

出版信息

J Physiol. 1995 Aug 15;487(1):67-90. doi: 10.1113/jphysiol.1995.sp020862.

Abstract
  1. We have used dendrite-attached patch-clamp techniques to record single Na+ and Ca2+ channel activity from the apical dendrites (up to 350 microns away from soma) of CA1 pyramidal neurons in rat hippocampal slices (ages: 2-8 weeks). 2. Na+ channels were found in every patch examined (range: 2 to > 20 channels per patch). Channel openings, which had a slope conductance of 15 +/- 0.3 pS (mean +/- S.E.M.), began with test commands to around -50 mV and consisted of both early transient channel activity and also later occurring prolonged openings of 5-15 ms. All Na+ channel activity was suppressed by inclusion of TTX (1 microM) in the recording pipette. 3. Ca2+ channel activity was recorded in about 80% of the patches examined (range: 1 to > 10 channels per patch). Several types of channel behaviour were observed in these patches. Single channel recordings in 110 mM BaCl2, revealed an approximately 10 pS channel of small unitary current amplitude (-0.5 pA at -20 mV). These channels began activating at relatively hyperpolarized potentials (-50 mV) and ensemble averages of this low voltage-activated (LVA) channel activity showed rapid inactivation. 4. A somewhat heterogeneous population of high voltage-activated, moderate conductance (HVAm; approximately 17 pS), Ca2+ channel activity was also encountered. These channels exhibited a relatively large unitary amplitude (-0.8 pA at 0 mV) and ensemble averages demonstrated moderate inactivation. The HVAm population of channels could be tentatively subdivided into two separate groups based upon mean channel open times. 5. Less frequently, HVA, large conductance (27 pS) Ca2+ channel activity (HVA1) was also observed. This large unitary amplitude (-1.5 pA at 0 mV) channel activity began with steps to approximately 0 mV and ensemble averages did not show any time-dependent inactivation. The dihydropyridine Ca2+ channel agonist Bay K 8644 (0.5 or 1 microM) was found to characteristically prolong these channel openings. 6. omega-Conotoxin MVIIC (10 microM), did not significantly reduce the amount of channel activity recorded from the LVA, HVAm or HVA1 channel types in dendritic patches. In patches from somata, omega-conotoxin MVIIC was effective in eliminating a significant amount of HVAm Ca2+ channel activity. Inclusion of 50 or 100 microM NiCl2 to the recording solution significantly reduced the amount of channel activity recorded from LVA and HVAm channel types in dendritic patches. A subpopulation of HVAm channels was, however, found to be Ni2+ insensitive. Dendritic HVA, channel activity was unaffected by these low concentrations of Ni2+.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 我们采用树突附着膜片钳技术,记录了大鼠海马脑片(年龄:2 - 8周)中CA1锥体神经元顶端树突(距胞体最远达350微米)的单个Na⁺和Ca²⁺通道活性。2. 在每个检测的膜片中均发现了Na⁺通道(每个膜片的通道数范围:2至超过20个)。通道开放时的斜率电导为15 ± 0.3 pS(平均值 ± 标准误),在测试指令达到约 -50 mV时开始,包括早期短暂的通道活性以及随后出现的持续5 - 15毫秒的长时间开放。所有Na⁺通道活性在记录电极内加入TTX(1微摩尔)后被抑制。3. 在约80%检测的膜片中记录到了Ca²⁺通道活性(每个膜片的通道数范围:1至超过10个)。在这些膜片中观察到了几种类型的通道行为。在110 mM BaCl₂中进行的单通道记录显示,有一个约10 pS的通道,其单通道电流幅度较小(在 -20 mV时为 -0.5 pA)。这些通道在相对超极化电位(-50 mV)时开始激活,这种低电压激活(LVA)通道活性的总体平均值显示快速失活。4. 还遇到了一群性质 somewhat heterogeneous 的高电压激活、中等电导(HVAm;约17 pS)的Ca²⁺通道活性。这些通道表现出相对较大的单通道幅度(在0 mV时为 -0.8 pA),总体平均值显示中等失活。基于平均通道开放时间,HVAm通道群体可初步分为两个不同的组。5. 较少见的是,还观察到了高电压激活、大电导(27 pS)的Ca²⁺通道活性(HVA1)。这种大的单通道幅度(在0 mV时为 -1.5 pA)通道活性在电位阶跃到约0 mV时开始,总体平均值未显示任何时间依赖性失活。发现二氢吡啶类Ca²⁺通道激动剂Bay K 8644(0.5或1微摩尔)可显著延长这些通道的开放时间。6. ω-芋螺毒素MVIIC(10微摩尔)并未显著减少从树突膜片中记录到的LVA、HVAm或HVA1通道类型的通道活性量。在胞体膜片中,ω-芋螺毒素MVIIC可有效消除大量的HVAm Ca²⁺通道活性。在记录溶液中加入50或100微摩尔NiCl₂可显著减少从树突膜片中记录到的LVA和HVAm通道类型的通道活性量。然而,发现HVAm通道的一个亚群对Ni²⁺不敏感。树突HVA通道活性不受这些低浓度Ni²⁺的影响。(摘要截取自400字)
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/296f/1156600/05ca3d11a0d8/jphysiol00311-0074-a.jpg

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