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非洲爪蟾卵母细胞兴奋性的诱导与消失:一项电压钳研究

Induction and disappearance of excitability in the oocyte of Xenopus laevis: a voltage-clamp study.

作者信息

Baud C, Kado R T

出版信息

J Physiol. 1984 Nov;356:275-89. doi: 10.1113/jphysiol.1984.sp015464.

Abstract

Electrically excitable, sodium-selective channels are induced in the membrane of the oocytes of Xenopus laevis when it is submitted to prolonged positive potentials (Kado, Marcher & Ozon, 1979; Baud, Kado & Marcher, 1982). Under a long positive voltage-clamp step, the membrane current, initially outward, becomes inward with a sigmoidal time course. The mean time to half-maximal inward current (t 1/2) is about 18 s at 16 degrees C when stepping the membrane potential to +55 mV. The rate of channel induction was very temperature dependent (Q10 about 5). In an Arrhenius plot, the t 1/2 for induction at temperatures between 5 and 22 degrees C showed a single slope. The rate of induction was dependent on the membrane potential, increasing exponentially with positive membrane potential (e-fold for a 20 mV change). When the membrane was maintained at resting potential after induction, the ability to produce inward currents with short depolarizing steps slowly disappeared with a t 1/2 of 4 min at 16 degrees C. The temperature dependence for disappearance was larger than that found for induction (Q10 about 7). The rate of disappearance was not dependent on holding the membrane potential in the range -30 to -100 mV. Induction proceeded in calcium-free medium. Cycloheximide, a potent protein synthesis inhibitor had no effect (100 micrograms/ml) on the induction rate. Isobutylmethylxanthine (IBMX) or theophyllin (phosphodiesterase inhibitors) applied externally (10(-4) M) did not affect the induction or disappearance rates. From the present results, mechanisms such as protein synthesis or a second messenger (such as calcium or cyclic AMP) do not appear to be involved. During the depolarization necessary to obtain induction, another conductance was also activated. It was more slowly established, appeared to be non-saturable and had a reversal potential between zero and -10 mV. It was found to be very much reduced at temperatures below about 16 degrees C.

摘要

当非洲爪蟾卵母细胞膜受到长时间的正电位作用时,可诱导出电兴奋性的、对钠有选择性的通道(Kado、Marcher和Ozon,1979年;Baud、Kado和Marcher,1982年)。在长时间的正电压钳制步骤下,膜电流最初是外向的,随后呈S形时间进程变为内向。当将膜电位钳制到+55 mV时,在16℃下达到内向电流最大值一半的平均时间(t1/2)约为18秒。通道诱导速率对温度非常敏感(Q10约为5)。在阿伦尼乌斯图中,5至22℃之间温度下诱导的t1/2呈现单一斜率。诱导速率取决于膜电位,随正膜电位呈指数增加(20 mV变化时增加一倍)。诱导后当膜维持在静息电位时,用短的去极化步骤产生内向电流的能力在16℃下以4分钟的t1/2缓慢消失。消失的温度依赖性大于诱导时的温度依赖性(Q10约为7)。消失速率不依赖于将膜电位保持在-30至-100 mV范围内。诱导在无钙培养基中进行。强力蛋白质合成抑制剂环己酰亚胺(100微克/毫升)对诱导速率无影响。外部施加的异丁基甲基黄嘌呤(IBMX)或茶碱(磷酸二酯酶抑制剂)(10-4 M)不影响诱导或消失速率。从目前的结果来看,蛋白质合成或第二信使(如钙或环磷酸腺苷)等机制似乎未参与其中。在获得诱导所需的去极化过程中,另一种电导也被激活。它建立得更慢,似乎是非饱和的,反转电位在零至-10 mV之间。发现在约16℃以下的温度下它大大降低。

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