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注射大鼠骨骼肌RNA的非洲爪蟾卵母细胞中的钙通道电流。

Calcium channel currents in Xenopus oocytes injected with rat skeletal muscle RNA.

作者信息

Dascal N, Lotan I, Karni E, Gigi A

机构信息

Department of Physiology and Pharmacology, Sackler Faculty of Medicine, Tel Aviv University, Ramat Aviv, Israel.

出版信息

J Physiol. 1992 May;450:469-90. doi: 10.1113/jphysiol.1992.sp019137.

DOI:10.1113/jphysiol.1992.sp019137
PMID:1279162
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1176132/
Abstract
  1. Ba2+ currents (IBa) through voltage-dependent Ca2+ channels were studied in Xenopus laevis oocytes injected with heterologous RNA extracted from skeletal muscle (SkM) of young rats, using the two-electrode voltage clamp technique. 2. With 40 or 50 mM-extracellular Ba2+, native oocytes of most frogs displayed IBa between -5 and -20 nA at 0 mV. However, in 'variant' native oocytes of four frogs, IBa exceeded -30 nA and reached up to -100 nA. In oocytes injected with SkM RNA, IBa of up to -250 nA was observed. 3. In SkM RNA-injected oocytes and 'variant' native oocytes, the decay of IBa displayed two kinetic components. The faster component was selectively blocked by 40-100 microM-Ni2+ and thus was termed the Ni(2+)-sensitive IBa. The slower component was Ni2+ resistant, being inhibited only 10-20% by 100-200 microM-Ni2+. The half-activation and the half-inactivation voltages of the Ni(2+)-sensitive IBa were more negative (by 14.5 and 28.7 mV, respectively) than those of the Ni(2+)-resistant IBa. 4. Neither Ni(2+)-sensitive nor Ni(2+)-resistant IBa in native or SkM RNA-injected oocytes were affected by dihydropyridine antagonists nifedipine and (+) PN 200-110 (1-10 microM), by the dihydropyridine agonist (-)Bay K 8644 (0.01-2 microM), or by verapamil below 50 microM. IBa was blocked by diltiazem (half-block at about 500 microM). Thus, the pharmacology of IBa in SkM RNA-injected and in native oocytes was not characteristic of the L-type Ca2+ channel abundant in the skeletal muscle. 5. Destruction of the RNA coding for the channel-forming alpha 1-subunit of the SkM L-type Ca2+ channel using a hybrid arrest method failed to selectively suppress the appearance of either Ni(2+)-sensitive or Ni(2+)-resistant IBa in SkM RNA-injected oocytes. 6. Our results suggest that the appearance of large voltage-dependent Ba2+ currents in SkM RNA-injected oocytes is not due to the expression of the alpha 1-subunit of the SkM L-type Ca2+ channel. The possibility that the expression of a channel-forming subunit of another Ca2+ channel type underlies one of these currents cannot be rejected. However, since the Ba2+ currents in SkM RNA-injected oocytes resemble those observed in native oocytes, we suggest that their appearance may be the result of an enhanced activity of the native Ca2+ channels, possibly due to the expression of the 'auxiliary' subunits of the SkM Ca2+ channel that form complexes with a native alpha 1-subunit.
摘要
  1. 采用双电极电压钳技术,在注射了从幼年大鼠骨骼肌(SkM)中提取的异源RNA的非洲爪蟾卵母细胞中,研究了通过电压依赖性Ca2+通道的Ba2+电流(IBa)。2. 细胞外Ba2+浓度为40或50 mM时,大多数青蛙的天然卵母细胞在0 mV时的IBa在-5至-20 nA之间。然而,在四只青蛙的“变异”天然卵母细胞中,IBa超过-30 nA,最高可达-100 nA。在注射了SkM RNA的卵母细胞中,观察到的IBa高达-250 nA。3. 在注射了SkM RNA的卵母细胞和“变异”天然卵母细胞中,IBa的衰减表现出两个动力学成分。较快的成分被40 - 100 μM的Ni2+选择性阻断,因此被称为Ni(2+)敏感的IBa。较慢的成分对Ni2+有抗性,100 - 200 μM的Ni2+仅抑制其10 - 20%。Ni(2+)敏感的IBa的半激活电压和半失活电压比Ni(2+)抗性的IBa更负(分别为14.5和28.7 mV)。4. 天然或注射了SkM RNA的卵母细胞中的Ni(2+)敏感或Ni(2+)抗性IBa,均不受二氢吡啶拮抗剂硝苯地平和(+) PN 200 - 110(1 - 10 μM)、二氢吡啶激动剂(-)Bay K 8644(0.01 - 2 μM)或低于50 μM的维拉帕米的影响。IBa被地尔硫䓬阻断(约500 μM时半阻断)。因此,注射了SkM RNA的卵母细胞和天然卵母细胞中IBa的药理学特性并非骨骼肌中丰富的L型Ca2+通道的特征。5. 使用杂交抑制法破坏SkM L型Ca2+通道形成通道的α1亚基的RNA,未能选择性抑制注射了SkM RNA的卵母细胞中Ni(2+)敏感或Ni(2+)抗性IBa的出现。6. 我们的结果表明,注射了SkM RNA的卵母细胞中出现大的电压依赖性Ba2+电流并非由于SkM L型Ca2+通道α1亚基的表达。不能排除另一种Ca2+通道类型的通道形成亚基的表达是其中一种电流的基础的可能性。然而,由于注射了SkM RNA的卵母细胞中的Ba2+电流与天然卵母细胞中观察到的电流相似,我们认为它们的出现可能是天然Ca2+通道活性增强的结果,可能是由于SkM Ca2+通道的“辅助”亚基与天然α1亚基形成复合物的表达所致。
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b7b1/1176132/7d2eac8b46fc/jphysiol00432-0482-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b7b1/1176132/7d2eac8b46fc/jphysiol00432-0482-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b7b1/1176132/7d2eac8b46fc/jphysiol00432-0482-a.jpg

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