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培养的鸡心脏细胞中的钠钙耦合转运

Coupled sodium-calcium transport in cultured chick heart cells.

作者信息

Murphy E, Wheeler D M, LeFurgey A, Jacob R, Lobaugh L A, Lieberman M

出版信息

Am J Physiol. 1986 Mar;250(3 Pt 1):C442-52. doi: 10.1152/ajpcell.1986.250.3.C442.

Abstract

In cultured embryonic chick heart cells, alterations of extracellular Na (Nao) and Ca (Cao), intracellular Na (Nai) and Ca, extracellular pH, and membrane potential resulted in changes in Na and Ca contents that were consistent with sarcolemmal Na-Ca exchange. 24Na efflux measurements revealed a large ouabain-insensitive component, one-third of which was inhibited by removal of Cao. Incubating the cells in Na-free solution resulted in a rapid, 1.5- to 2-fold increase in total cell Ca that remained elevated for at least 15 min. Cells exposed for 15 min to Nao less than or equal to 20 mM became maximally loaded with Ca, whereas Ca loading fell off sharply at values of Nao greater than 20 mM. The movement of Na against its electrochemical gradient was shown to be associated with Ca accumulation. During Na-K pump inhibition (in 10(-4) M ouabain), Na initially rose 2- to 3-fold to a level below its equilibrium value; then, lowering Cao for 30 min from 1.25 to 0.75 mM caused a 26% elevation in Nai, whereas raising Cao from 1.25 to 2.7 mM resulted in a 25% fall in Nai against its electrochemical gradient. These data are consistent with Nai being maintained by a Na-Ca exchange during Na-K pump inhibition. In the presence of ouabain (10(-4) M), Ca uptake into intracellular organelles, e.g., mitochondria, was suggested by an increase in total cell Ca as well as the occurrence of mitochondrial matrix granules, which were shown qualitatively by X-ray analysis to contain Ca. Although matrix granules also occurred in mitochondria during Na-free incubation, they did not contain detectable amounts of Ca when examined under identical conditions of fixation and analysis.

摘要

在培养的鸡胚心脏细胞中,细胞外钠(Nao)和钙(Cao)、细胞内钠(Nai)和钙、细胞外pH值以及膜电位的改变导致钠和钙含量发生变化,这些变化与肌膜钠-钙交换一致。24Na外流测量显示存在一个大的哇巴因不敏感成分,其中三分之一可通过去除Cao来抑制。将细胞置于无钠溶液中孵育会导致细胞总钙迅速增加1.5至2倍,并至少保持升高15分钟。暴露于Nao小于或等于20 mM的细胞15分钟后会达到最大钙负载,而当Nao大于20 mM时,钙负载量会急剧下降。钠逆其电化学梯度的移动与钙积累相关。在钠-钾泵抑制期间(在10(-4) M哇巴因中),钠最初升高2至3倍至低于其平衡值的水平;然后,将Cao从1.25 mM降低至0.75 mM 30分钟会导致Nai升高26%,而将Cao从1.25 mM升高至2.7 mM会导致Nai逆其电化学梯度下降25%。这些数据与在钠-钾泵抑制期间通过钠-钙交换维持Nai一致。在存在哇巴因(10(-4) M)的情况下,细胞总钙增加以及线粒体基质颗粒的出现提示钙被摄取到细胞内细胞器(如线粒体)中,通过X射线分析定性显示这些颗粒含有钙。尽管在无钠孵育期间线粒体中也出现了基质颗粒,但在相同的固定和分析条件下检查时,它们不含可检测量的钙。

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