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在非洲爪蟾卵母细胞中,由受体激活、三磷酸肌醇和钙所诱发的膜电流潜伏期。

Latencies of membrane currents evoked in Xenopus oocytes by receptor activation, inositol trisphosphate and calcium.

作者信息

Miledi R, Parker I

机构信息

Department of Psychobiology, University of California Irvine 92717.

出版信息

J Physiol. 1989 Aug;415:189-210. doi: 10.1113/jphysiol.1989.sp017718.

DOI:10.1113/jphysiol.1989.sp017718
PMID:2484206
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1189173/
Abstract
  1. Application of serum to Xenopus oocytes elicits an oscillatory chloride membrane current, which begins after a latency of several seconds or minutes, and is mediated through a phosphoinositide-calcium signalling pathway. We studied the characteristics and origin of this latency in voltage-clamped oocytes. 2. Bath application of low doses of serum evoked responses beginning after latencies of 1 min or more. The latency decreased with increasing dose and reached a minimal value of several seconds that did not decrease with further increases in serum concentration. Experiments to study this minimal latency were done by applying brief 'puffs' of serum and other agonists at high concentrations from a local extracellular pipette. 3. The mean latency of the response evoked by local serum application was about 7 s (at 22-24 degrees C), but individual responses showed a wide variation, from 2 s to over 20 s. Diffusion of serum from the pipette tip to the membrane did not contribute appreciably to this delay, since short (less than 100 ms) delays were obtained when KCl was applied in the same way. 4. Currents evoked by acetylcholine and serotonin, in oocytes induced to acquire muscarinic and serotonergic receptors following injection of brain messenger RNA, began following latencies similar to that of the serum response. 5. The response latency was shorter when serum was applied to the vegetal rather than the animal hemisphere of the oocyte, even though smaller currents were obtained. 6. The latency showed a slight dependence upon membrane potential, becoming shorter with depolarization. 7. Cooling to temperatures below about 22 degrees C produced a striking lengthening of the delay, corresponding to a Q10 of about 5. In contrast, above 22 degrees C the temperature dependence was slight, with a Q10 of about 1.25. 8. Intracellular injections of calcium and inositol 1,4,5-trisphosphate (IP3) evoked chloride currents with short (a few tens of milliseconds) latency. Short (100 ms) latency responses were also evoked when intracellularly loaded caged IP3 was photolysed by strong illumination, but weak illumination gave responses with latencies of over 1 min. 9. Measurements of intracellular free calcium, made with Fura-2 and Indo-1, showed an increase following serum application beginning coincident with the onset of the membrane current response.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 将血清应用于非洲爪蟾卵母细胞会引发振荡性氯离子膜电流,该电流在几秒或几分钟的延迟后开始,并通过磷脂酰肌醇 - 钙信号通路介导。我们在电压钳制的卵母细胞中研究了这种延迟的特征和起源。2. 低剂量血清的浴槽应用引发的反应在1分钟或更长时间的延迟后开始。延迟随剂量增加而减小,并达到几秒的最小值,且随着血清浓度进一步增加不再减小。通过从局部细胞外移液管以高浓度施加血清和其他激动剂的短暂“吹入”来进行研究这种最小延迟的实验。3. 局部应用血清引发反应的平均延迟约为7秒(在22 - 24摄氏度),但个体反应显示出很大差异,从2秒到超过20秒。血清从移液管尖端扩散到膜对这种延迟的贡献不大,因为以相同方式施加氯化钾时获得的延迟很短(小于100毫秒)。4. 在注射脑信使核糖核酸后诱导获得毒蕈碱和5 - 羟色胺能受体的卵母细胞中,乙酰胆碱和5 - 羟色胺引发的电流在类似于血清反应的延迟后开始。5. 将血清应用于卵母细胞的植物半球而非动物半球时,反应延迟更短,尽管获得的电流较小。6. 延迟对膜电位有轻微依赖性,随着去极化而变短。7. 冷却至约22摄氏度以下会使延迟显著延长,对应约5的Q10值。相比之下,在22摄氏度以上温度依赖性较小,Q10约为1.25。8. 细胞内注射钙和肌醇1,4,5 - 三磷酸(IP3)会引发延迟很短(几十毫秒)的氯离子电流。当通过强光照射使细胞内加载的笼锁IP3光解时,也会引发短(100毫秒)延迟反应,但弱光照射会产生延迟超过1分钟的反应。9. 用Fura - 2和Indo - 1进行的细胞内游离钙测量显示,血清应用后细胞内游离钙增加,与膜电流反应开始同时出现。(摘要截断于400字)