Endogenous ATP release inhibits electrogenic Na+ absorption and stimulates Cl− secretion in MDCK cells
Endogenous ATP release inhibits electrogenic Na+ absorption and stimulates Cl− secretion in MDCK cells
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MDCK 细胞中内源 ATP 释放抑制电 Na+ 吸收并刺激 Cl− 分泌
DOI：
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发表时间：
2007
期刊：
影响因子：
--
通讯作者：
J. Schafer
中科院分区：
文献类型：
--
作者：
Yi Xie;J. Schafer
我们以前对标记α，β的狗肾细胞系(FL-MDCK)和上皮钠通道γ亚基的研究表明，尽管大多数短路电流(ISC)是阿米洛利敏感的(AS-ISC)，但也有一个阿米洛利不敏感的成分(NS-ISC)是由于Cl−的分泌(Morris and Schafer，J Gen Physiol120：71-85,2002)。在目前的研究中，我们观察到在实验过程中NS-ISC逐渐增加，而AS-ISC相应减少。无论有无环磷酸腺苷(CAMP)治疗，AS-ISC与NS-ISC均呈显著负相关。NS-ISC可归因于囊性纤维化跨膜电导调节剂和4，4‘-二异硫氰基-2，2’-二磺酸二苯乙烯(DIDS)敏感的Ca~(2+)激活的Cl-−通道。在Ussing小室两侧持续灌流新鲜的浴液而不是再循环的浴液，或加入己糖激酶(6单位/ml)，可防止这种时间依赖性的改变，并使AS-ISC增加40-60%，而NS-ISC则成比例减少。在鲁米洛利存在的情况下，加入100gμM三磷酸腺苷可使NS-ISC一过性增加四倍，随后在基础水平上持续增加50%-60%。通过生物发光测量，单层细胞的ATP释放被发现发生在根尖而不是基底外侧膜上，并且cAMP处理使根尖释放增加了两倍。这些数据表明，在基础和cAMP刺激的条件下，结构性顶端三磷酸腺苷的释放是FL-−细胞ClATP分泌的时间依赖性增加和ENaC介导的Na+吸收的比例下降的基础。因此，内源性三磷酸腺苷的释放可以在与该上皮细胞和类似的上皮细胞的实验中引入一个显著的混杂变量，它可能至少是所观察到的Cl-−分泌和Na+吸收之间的一些相互作用的基础。
Our previous studies with a line of Madin-Darby canine kidney (MDCK) cells (FL-MDCK) transfected with FLAG-labeled α, β, and γ subunits of epithelial Na+ channel (ENaC) showed that, although most of the short-circuit current (Isc) was amiloride sensitive (AS-Isc), there was also an amiloride-insensitive component (NS-Isc) due to Cl− secretion (Morris and Schafer, J Gen Physiol 120:71–85, 2002). In the present studies, we observed a progressive increase in NS-Isc and a corresponding decrease in AS-Isc during experiments. There was a significant negative correlation between AS-Isc and NS-Isc both in the presence and absence of treatment with cyclic adenosine monophosphate (cAMP). NS-Isc could be attributed to both cystic fibrosis transmembrane conductance regulator (CFTR) and a 4, 4'-diisothiocyano-2, 2'-disulfonic acid stilbene (DIDS)-sensitive Ca2+-activated Cl− channel (CaCC). Continuous perfusion of both sides of the Ussing chamber with fresh rather than recirculated bathing solutions, or addition of hexokinase (6 U/ml), prevented the time-dependent changes and increased AS-Isc by 40–60%, with a proportional decrease in NS-Isc. Addition of 100 μM adenosine triphosphate (ATP) in the presence of luminal amiloride produced a transient four-fold increase in NS-Isc that was followed by a sustained increase of 50–60% above the basal level. ATP release from the monolayers, measured by bioluminescence, was found to occur across the apical but not the basolateral membrane, and the apical release was tripled by cAMP treatment. These data show that constitutive apical ATP release, which occurs under both basal and cAMP-stimulated conditions, underlies the time-dependent rise in Cl− secretion and the proportional fall in ENaC-mediated Na+ absorption in FL-MDCK cells. Thus, endogenous ATP release can introduce a significant confounding variable in experiments with this and similar epithelial cells, and it may underlie at least some of the observed interaction between Cl− secretion and Na+ absorption.
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DOI：
10.1152/ajprenal.1995.268.5.f854
发表时间：
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期刊：
The American journal of physiology.
影响因子：
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作者：
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DOI：
10.1152/ajprenal.1995.268.4.f671
发表时间：
1995
期刊：
The American journal of physiology.
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DOI：
10.1152/ajpcell.1999.277.2.c205
发表时间：
1999
期刊：
The American journal of physiology
影响因子：
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作者：
Luo,X;Zheng,W;Yan,M;Lee,MG;Muallem,S
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Muallem,S
DOI：
10.1152/ajplung.1996.271.2.l192
发表时间：
1996-08-01
影响因子：
4.9
作者：
Schultz, BD;DeRoos, ADG;Bridges, RJ
通讯作者：
Bridges, RJ