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利用镧来估算豚鼠结肠带中细胞外阳离子交换位点的数量及其对跨膜单价离子运动的影响。

The use of lanthanum to estimate the numbers of extracellular cation-exchanging sites in the guinea-pig's taenia coli, and its effects on transmembrane monovalent ion movements.

作者信息

Brading A F, Widdicombe J H

出版信息

J Physiol. 1977 Apr;266(2):255-73. doi: 10.1113/jphysiol.1977.sp011767.

Abstract
  1. Tissues were allowed to equilibrate in a Tris-buffered Krebs solution and were then exposed to similar solutions containing up to 5 mM-La. La caused shrinkage and significant losses of tissue K, Na, Mg and Ca. The shrinkage was exactly accountable for by a reduction in the extracellular space (e.c.s.) as measured by [14C]sucrose. No significant change was seen in tissue Cl content. Exposure for 1 hr to 5 mM-La led to a total cation loss of 24-3 +/- 1-6 m-equiv/kg or, correcting for the small change in Cl content, a loss of positive charge of 23-8 +/- 2-2 m-equiv/kg fresh wt. 2. Using the radioisotope 140La it was shown that this loss of cation was balanced by an uptake of La3+. 3. Subtraction of the ions in the measured [14C]sucrose space from the total tissue ion contents led to estimates of the "cellular" ion contents. The effects of 1 hr exposure to 5 mM-La on these were a loss of 12-9 +/- 2-4 m-equiv/kg of cation and a gain of 10-0 +/- 2-6 m-equiv/kg of Cl. 4. Similar changes in ion content were produced by La on "Na-loaded" and "K-loaded" tissues, these being tissues which by exposure to K-free or Na-free (high K) solutions had replaced all their K with Na or vice versa. 5. The uptakes of 24Na and 36Cl by Na-loaded tissues were both describable as the sum of two exponentail processes: a fast component (t 1/2 congruent to 1/2 min), which was presumed to be extracellular and a slower, presumed transmembrane, component. La reduced the rapid component of uptake of 24Na by an amount greater than that predicted by the reduction in the e.c.s., the extra amount lost being some 10--15 m-equiv/kg. La also reduced the amount of rapidly exchanging 36Cl, bwt this reduction was entirely accounted for by the change in the e.c.s. La reduced the rate constant of the slow component of 24Na uptake. 6. La reduced the rapidly exchanging component of 42K uptake by normal tissues by an amount equivalent to about 0-5 m-mole/kg fresh wt. of K in excess of the change in the extracellular space. 7. La had little effect on the effluxes of 36Cl and 42K from normal tissues. However, it reduced the size of the fastest component of exchange of 42K efflux from K-loaded tissues by an amount equal to some 10-15 m-equiv/kg in excess of the reduction in the e.c.s. A similar reduction in the rapidly exchanging component of 24Na efflux from normal tissues was also seen. La slowed the efflux of 24Na from Na-loaded tissues at times when the tracer lost could safely be regarded as intracellular. 8. The taenia coli when exposed to K-free solutions gains Na and loses K. In the presence of La the gain in Na was completely blocked. K was still lost, however, being accompanied by Cl and increased shrinkage. La also prevented the uptake of Na from high Na media by ion-depleted tissues (produced by exposure to sucrose media), while having little effect on the uptake of K from high K media by such tissues. 9 Tedia by such tissues. 9. The cation displaced by La in excess of that lost due to the reduction of the e.c.s...
摘要
  1. 将组织置于Tris缓冲的Krebs溶液中平衡,然后暴露于含有高达5 mM镧(La)的类似溶液中。La导致组织收缩,并使组织中的钾(K)、钠(Na)、镁(Mg)和钙(Ca)显著流失。收缩完全是由于细胞外间隙(e.c.s.)减小所致,这是通过[14C]蔗糖测量的。组织中的氯(Cl)含量未见明显变化。暴露于5 mM-La 1小时导致总阳离子流失24 - 3±1 - 6 m-equiv/kg,或者校正Cl含量的微小变化后,新鲜组织重量的正电荷流失为23 - 8±2 - 2 m-equiv/kg。

  2. 使用放射性同位素140La表明,这种阳离子流失被La3+的摄取所平衡。

  3. 从总组织离子含量中减去测量的[14C]蔗糖空间中的离子,可得出“细胞”离子含量的估计值。暴露于5 mM-La 1小时对这些离子的影响是阳离子流失12 - 9±2 - 4 m-equiv/kg,Cl增加10 - 0±2 - 6 m-equiv/kg。

  4. La对“钠负载”和“钾负载”组织的离子含量产生类似变化,这些组织通过暴露于无钾或无钠(高钾)溶液,已将其所有的K替换为Na,反之亦然。

  5. 钠负载组织对24Na和36Cl的摄取都可描述为两个指数过程的总和:一个快速成分(t 1/2约为1/2分钟),推测为细胞外成分,以及一个较慢的、推测为跨膜的成分。La使24Na摄取的快速成分减少的量大于根据e.c.s.减小所预测的量,额外损失的量约为10 - 15 m-equiv/kg。La也减少了快速交换的36Cl的量,但这种减少完全是由e.c.s.的变化引起的。La降低了24Na摄取慢成分的速率常数。

  6. La使正常组织中42K摄取的快速交换成分减少的量相当于约0 - 5 m-mole/kg新鲜组织重量的K,超过细胞外空间的变化量。

  7. La对正常组织中36Cl和42K的外流影响很小。然而,它使钾负载组织中42K外流最快交换成分的大小减少,减少量超过e.c.s.减小量约10 - 15 m-equiv/kg。正常组织中24Na外流的快速交换成分也有类似减少。当示踪剂损失可安全地视为细胞内时,La减缓了钠负载组织中24Na的外流。

  8. 结肠带暴露于无钾溶液时会摄取Na并丢失K。在有La存在时,Na的摄取被完全阻断。然而,K仍会丢失,并伴有Cl和收缩增加。La还阻止了离子耗尽组织(通过暴露于蔗糖培养基产生)从高钠培养基中摄取Na,而对这种组织从高钾培养基中摄取K影响很小。

  9. La置换的阳离子超过因e.c.s.减小而丢失的阳离子……

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