Lymphocyte membrane potential and Ca2+‐sensitive potassium channels described by oxonol dye fluorescence measurements
Lymphocyte membrane potential and Ca2+‐sensitive potassium channels described by oxonol dye fluorescence measurements
复制标题
通过 oxonol 染料荧光测量描述淋巴细胞膜电位和 Ca2+ 敏感钾通道
DOI：
--
复制
发表时间：
1985
影响因子：
5.6
通讯作者：
T. Chused
中科院分区：
文献类型：
--
作者：
H. A. Wilson;T. Chused
本文描述了一种通过流式细胞术记录单细胞的氧醇染料荧光定量测量淋巴细胞跨膜电位差（Ψ）的方法。同时收集和分析多个光学参数以及使用带负电荷的oxonol探针可以比采用阳离子电压指示器的体电池悬浮技术更准确地测量Ψ。小鼠脾脏和人血淋巴细胞Ψ计算为−70 mV。T淋巴细胞和B淋巴细胞保持恒定Ψ，因为细胞外K+从2到10 mM变化，偏离K+平衡电位（Ek）显示是由Na+渗透性引起的。当[K+]o值大于10 mM时，淋巴细胞表现为K+电极。淋巴细胞亚群检测显示，Ca2+离子载体A23187诱导的超极化仅发生在T细胞中。这种反应被确定为通过药理学操作激活Ca2+敏感的K+通道。因此，观察到通过4 -氨基吡啶（4 - AP, 10 mM）去极化的T细胞通过A23187诱导的[Ca2+]i升高Ψ回到静止状态。A23187对奎宁（100 μM）去极化细胞无影响。在T细胞中，Ca2+激活通道不影响静息Ψ，因为它可以被奎宁（20 μM）选择性阻断或被钙调素拮抗剂（5 μM三氟拉嗪）调节而不影响静息Ψ。
A method is described for quantitative measurement of lymphocyte transmembrane electrical potential difference (Ψ) by flow cytometric recording of the oxonol dye fluorescence of single cells. Both the simultaneous collection and analysis of multiple optical parameters and the use of a negatively charged oxonol probe allowed more accurate measurement of Ψ than may be obtained by bulk cell suspension techniques employing cationic voltage indicators. Mouse spleen and human blood lymphocyte Ψ was calculated to be −70 mV. T and B lymphocytes maintain a constant Ψ as extracellular K+ is varied from 2 to 10 mM and the deviation from K+ equilibrium potentials (Ek) is shown to result from Na+ permeability. At [K+]o values greater than 10 mM, lymphocytes behave as K+ electrodes. Examination of lymphocyte subsets showed that hyperpolarization induced by the Ca2+ ionophore A23187 occurs only in T cells. This response was identified as activation of a Ca2+‐sensitive K+ channel by pharmacologic manipulations. Hence, T cells depolarized by 4‐aminopyridine (4‐AP, 10 mM) were observed to return to resting Ψ by A23187‐induced elevation of [Ca2+]i. Cells depolarized by quinine (100 μM) were unaffected by A23187. The Ca2+‐activated channel does not contribute to resting Ψ in T cells since it may be selectively blocked by quinine (20 μM) or modulated by calmodulin antagonists (5 μM trifluperazine) without affecting resting Ψ.
影响因子：
64.8
作者：
DECOURSEY, TE;CHANDY, KG;CAHALAN, MD
通讯作者：
CAHALAN, MD
纯化的小鼠 B 淋巴细胞在无血清成分确定的培养基中产生体外初级抗体反应。
DOI：
--
发表时间：
1981
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Mosier,DE
通讯作者：
Mosier,DE