• The Korean Journal of Physiology and Pharmacology
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• 제11권5호
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• pp.183-188
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• 2007

Using BHK cells with stable expression of cardiac $Na^+/Ca^{2+}$ exchanger(BHK-NCX1), reverse mode(i.e. $Ca^{2+}$ influx mode) of NCX1 current was recorded by whole-cell patch clamp. Repeated stimulation of reverse NCX1 produced a cytosolic $Ca^{2+}$-dependent long-term inactivation of the exchanger activity. The degrees of inactivation correlated with NCX1 densities of the cells and were attenuated by reduced $Ca^{2+}$ influx via the reverse exchanger. The inactivation of NCX1 was attenuated by(i) inhibition of $Ca^{2+}$ influx with reduced extracellular $Ca^{2+}$, (ii) treatment with NCX1 blocker($Na^{2+}$), and (iii) increase of cytoplasmic $Ca^{2+}$ buffer(EGTA). In BHK-NCX1 cells transiently expressing TRPV1 channels, $Ca^{2+}$ influx elicited by capsaicin produced a marked inactivation of NCX1. We suggest that cytoplasmic $Ca^{2+}$ has a dual effect on NCX1 activities, and that allosteric $Ca^{2+}$ activation of NCX1 can be opposed by the $Ca^{2+}$-dependent long-term inactivation in intact cells.

• BMB Reports
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• 제28권6호
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• pp.499-503
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• 1995

In our previous studies (Chae et al., 1990; Chae et a1., 1993), we found that a phytochrome signal was clearly connected with the change in cytosolic free $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) in oat cells. It was determined that the $[Ca^{2+}]_i$ change occured both by mobilization out of the intracellular $Ca^{2+}$ store and by influx from the medium. The specific aim of this work is to elucidate the processes connecting $Ca^{2+}$ mobilization and influx. The cells treated with thapsigargin (increasing $[Ca^{2+}]_i$ by inhibition of the $Ca^{2+}$-ATPase in the calcium pool) in the presence of external $Ca^{2+}$ showed the same increasing pattern (sustained increasing shape) of $[Ca^{2+}]_i$ as that measured in animal cells. Red light irradiation after thapsigargin treatment did not increase $[Ca^{2+}]_i$ These results suggest that thapsigargin also acts specifically in the processes of mobilization and influx of $Ca^{2+}$ in oat cells. When the cells were treated with TEA ($K^+$ channel blocker), changes in $[Ca^{2+}]_i$ were drastically reduced in comparison with that measured in the absence of TEA. The results suggest that the change in $[Ca^{2+}]_i$ due to red light irradiation is somehow related with $K^+$ channel opening to change membrane potential. The membrane potential change due to $K^+$ influx might be the critical factor in opening a voltage-dependent calcium channel for $Ca^{2+}$ influx.

• 동의생리병리학회지
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• 제18권5호
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• pp.1382-1386
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• 2004

This study was performed for the investigation of vasodilatory efficacy and its underlying mechanisms of Samhwangsasim-tang(SST), herbal remedy. SST relaxed vascular strips precontracted with phenylephrine or KCI(51 mM), but the magnitude of relaxation was greater in phenylephrine(PE) induced contraction. The relaxation effects of SST was endothelium-independent. L-NAME, iNOS inhibitor, and methyl en blue(MB), cGMP inhibitor, did not attenuate the relaxation responses of SST. In the absence of extracellular Ca2+, pre-incubation of the aortic rings with SST significantly reduced the contraction by PE, suggesting that the relaxant action of the SST includes inhibition of Ca/sup 2+/ influx and release of Ca/sup 2+/ from intracellular stores (SR). In addition, the cell death was induced by SST in human aortic smooth muscle cells but not that of human umbilical vein endothelial cells. We conclude that in rat thoracic aorta, SST may induce in part vasodilation through inhibition of Ca/sup 2+/ influx and release of Ca/sup 2+/ from intracellular stores.

• The Korean Journal of Physiology and Pharmacology
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• 제2권1호
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• pp.109-117
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• 1998

Role of $Ca^{2+}$/calmodulin complex in intracellular $Ca^{2+}$ mobilization in neutrophils has not been clearly elucidated. In this study, effects of chlorpromazine, trifluoperazine and imipramine on the intracellular $Ca^{2+}$ mobilization, including $Ca^{2+}$ influx, in C5a-activated neutrophils were investigated. Complement C5a- stimulated superoxide production and myeloperoxidase release in neutrophils were inhibited by chlorpromazine, trifluoperazine and imipramine, except no effect of imipramine on myeloperoxidase release. A C5a-elicited elevation of [$Ca^{2+}$]i in neutrophils was inhibited by chlopromazine, trifluoperazine, imipramine, staurosporine, genistein, EGTA, and verapamil but not affected by pertussis toxin. The intracellular $Ca^{2+}$ release in C5a-activated neutrophils was not affected by chlorpromazine and imipramine. Chlorpromazine and imipramine inhibited $Mn^{2+}$ influx by C5a-activated neutrophils. Thapsigargin-evoked $Ca^{2+}$ entry was inhibited by chlorpromazine, trifluoperazine, imipramine, genistein, EGTA and verapamil, while the effect of staurosporine was not detected. The results suggest that $Ca^{2+}$/calmodulin complex is involved in the activation process of neutrophils. The depressive action of calmodulin inhibitors on the elevation of cytosolic $Ca^{2+}$ level in C5a-activated neutrophils appears to be accomplished by inhibition of $Ca^{2+}$ influx from the extracellular medium.

• The Korean Journal of Physiology
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• 제18권2호
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• pp.125-137
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• 1984

Vanadate의 수축에 이용되는 $Ca^{2+}$의 동원 경로와 Na-Pump억제가 vanadate의 수축에 어떤 영향을 미치는 지를 밝히기 위해 본 실험을 시행하여 다음과 같은 곁과를 얻었다. 1) 흰쥐의 자궁근에서는 vanadate는 수축을 일으켜 $5{\times}10^{-4}M$에서 최대수축을 나타내었으며 사람의 자궁근이 흰쥐의 자중근에 비해 vanadate에 더 민감한 반응을 보였다. 2) Vanadate에 의한 수축은 $Ca^{2+}$제거에 의해 완전히 억제되지 않았고 사람의 자궁근이 외부 $Ca^{2+}$의 농도변화에 더 민감한 반응을 보였다. 3) Vanadate에 의한 수축은 verapamil농도를 증가시킴에 따 억제되었으며 100k에 극한 수축을 완전 억제시키는$3{\times}10^{-5}M$ verapamil 존재하에서도 최대수의 40%정도가 남아있었고, 이 크기는 $Ca^{2+}$없는 용액에서의 수축의 크기와 유사하였다. 4) Na-pump억제시 vanadate의 수축은 증가하였고 이 현상은 $3{\times}10^{-5}M$ verapamil 존재하에서도 나타났다. 5) $Ca^{2+}$없는 ouabain용액에서 전처치후에 vanadate에 의한 수축은 증가하지 않았으나 외부내 $Ca^{2+}$을 부가할 나타나는 반음은 대조군에 비해 현저히 증가하였다. 6) Verapamil 존재시 vanadate에 의한 $Ca^{45}$유입은 완전히 억제되었으나 ouabain으로 처리한 후는 verapamil 존재하에서도 vanadate가 현저히 $Ca^{45}$유입을 일으켰다. 7) Ouabain이나 K 없는 용액으로 치리시간이 증가함에 따라 vanadate에 의한 수축의 증가정도는 더욱 더 현저하였다. 8) Ouabain 전처치시 증가된 vanadate에 의한 수축은 $10^{-4}M$ papaverine에 의해 현저히 억제되었다. 9) Acetylcholine에 의한 수축은 verapamil 존재하에서도 Na-pump억제 시간이 증가함에 따라 증가하였다. 이상의 결과로 볼 때 vanadate에 대해 사람의 자궁근이 흰쥐의 자궁근에 비해 더 민감한 반응을 보이고 vanadate에 의한 수축에는 외부와 내부 $Ca^{2+}$이 모두 이용되며 Na-pump 억제시 여러가지 근수축물질이 verapamil에 의해 억제되지 않는 $Ca^{2+}$유입을 일으키며 이 유입경로의 성질은 확실히 알 수 없으나 Papaverine에 의해 억제되며 막전위의 변화와 관련이 있는 것으로 생각된다.

• Natural Product Sciences
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• 제12권4호
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• pp.217-220
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• 2006

The effects of $(1R,9S)-\beta-hydrastine$ hydrochloride (BHSH) on $Ca^{2+}$ transport in rat pheochromocytoma PC12 cells were investigated. In the presence of external $Ca^{2+}$, BHSH at $100{\mu}M$ inhibited $K^+$ (56mM)-induced dopamine release, and $K^+-induced$ $Ca^{2+}$ influx and a sustained rise of $[Ca^{2+}]_i$. In addition, BHSH at 100 f.!M reduced the sustained rise of $[Ca^{2+}]_i$ elicited by 20 mM caffeine, but not by $1{\mu}M$ thapsigargin, in presence of external $Ca^{2+}$. These results suggest that BHSH inhibited $K^+-induced$ dopamine release and $[Ca^{2+}]_i$ influx, and store-operated $Ca^{2+}$ channels activated by caffeine, but not by thapsigargin, in PC12 cells.

• Archives of Pharmacal Research
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• 제28권6호
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• pp.709-715
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• 2005

The purpose of this study was to investigate the effect of atropine on peripheral vasodilation and the mechanisms involved. The isometric tension of rat mesenteric artery rings was recorded in vitro on a myograph. The results showed that atropine, at concentrations greater than 1$\mu$M, relaxed the noradrenalin (NA)-precontracted rat mesenteric artery in a concentration-dependent manner. Atropine-induced vasodilatation was mediated, in part, by an endothelium-dependent mechanism, to which endothelium-derived hyperpolarizing factor may contribute. Atropine was able to shift the NA-induced concentration-response curve to the right, in a non-parallel manner, suggesting the mechanism of atropine was not mediated via the ${\alpha}_1$-adrenoreceptor. The $\beta$-adrenoreceptor and ATP sensitive potassium channel, a voltage dependent calcium channel, were not involved in the vasodilatation. However, atropine inhibited the contraction derived from NA and $CaCl_2$ in $Ca^{2+}$-free medium, in a concentration dependent manner, indicating the vasodilatation was related to the inhibition of extracellular $Ca^{2+}$ influx through the receptor-operated calcium channels and intracellular $Ca^{2+}$ release from the $Ca^{2+}$ store. Atropine had no effect on the caffeine-induced contraction in the artery segments, indicating the inhibition of intracellular $Ca^{2+}$ release as a result of atropine most likely occurs via the IP3 pathway rather than the ryanodine receptors. Our results suggest that atropine-induced vasodilatation is mainly from artery smooth muscle cells due to inhibition of the receptor-mediated $Ca^{2+}$-influx and $Ca^{2+}$-release, and partly from the endothelium mediated by EDHF.

• Molecules and Cells
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• 제19권2호
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• pp.268-278
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• 2005

We investigated the effect of cytosolic and extracellular $Ca^{2+}$ on $Ca^{2+}$ signals in pancreatic acinar cells by measuring $Ca^{2+}$ concentration in the cytosol($[Ca^{2+}]_c$) and in the lumen of the ER($[Ca^{2+}]_{Lu}$). To control buffers and dye in the cytosol, a patch-clamp microelectrode was employed. Acetylcholine released $Ca^{2+}$ mainly from the basolateral ER-rich part of the cell. The rate of $Ca^{2+}$ release from the ER was highly sensitive to the buffering of $[Ca^{2+}]_c$ whereas ER $Ca^{2+}$ refilling was enhanced by supplying free $Ca^{2+}$ to the cytosol with $[Ca^{2+}]_c$ clamped at resting levels with a patch pipette containing 10 mM BAPTA and 2 mM $Ca^{2+}$. Elevation of extracellular $Ca^{2+}$ to 10 mM from 1 mM raised resting $[Ca^{2+}]_c$ slightly and often generated $[Ca^{2+}]_c$ oscillations in single or clustered cells. Although pancreatic acinar cells are reported to have extracellular $Ca^{2+}$-sensing receptors linked to phospholipase C that mobilize $Ca^{2+}$ from the ER, exposure of cells to 10 mM $Ca^{2+}$ did not decrease $[Ca^{2+}]_{Lu}$ but rather raised it. From these findings we conclude that 1) ER $Ca^{2+}$ release is strictly regulated by feedback inhibition of $[Ca^{2+}]_c$, 2) ER $Ca^{2+}$ refilling is determined by the rate of $Ca^{2+}$ influx and occurs mainly in the tiny subplasmalemmal spaces, 3) extracellular $Ca^{2+}$-induced $[Ca^{2+}]_c$ oscillations appear to be triggered not by activation of extracellular $Ca^{2+}$-sensing receptors but by the ER sensitised by elevated $[Ca^{2+}]_c$ and $[Ca^{2+}]_{Lu}$.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.97.2-97.2
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• 2003

(1R,9S)-(${\beta}$)-Hydrastine (HS) at 10-50 ${\mu}$M has been proven to have an inhibitory effect on dopamine biosynthesis in PC12 cells by the inhibition of tyrosine hydroxylase (TH) activity and TH gene expression. In the present study, therefore, the effects of HS on the basal and K$\^$+/-induced dopamine release, and Ca$\^$2+/ influx induced by high K$\^$+/ and caffeine in PC12 cells were investigated. The dopamine release by high K$\^$+/ (56 mM) was inhibited by co-incubation of 20 ${\mu}$M HS. Application of HS also significantly reduced the magnitude of the maintained Ca$\^$2+/ influx induced by K$\^$+/ depolarization. (omitted)

• The Korean Journal of Ecology
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• 제20권2호
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• pp.83-94
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• 1997

Stomatal closing by ozone and water stress could reduce further ozone injury by inhibition of ozone influx to the tissue. Direct effect of ozone on stomata can be explained from two aspects which are a stimulation of stomatal closing and an inhibition of stomatal opening. An increase of $Ca^{2+}$ influx into cytoplasm by ozone could stimulate potassium efflux ion channel and inhibits inward potassium ion channels. By this mechanism ozone could induce stomatal closing. On the other hand, ozone could inhibit stomatal opening by affecting the activity of $H^{+}$ dependent ATPase of the membrane in guard cells. This would inhibit proton efflux which precede stomatal opening. It is also possible that ozone could reduce the activity of photosynthesis in guard cells which lead to affect the production of osmotically active sugars and energy. Indirect effect of ozone to stomata is through the effect of $CO_2$ elevation as a result of damage of the photozynthetic machinery. This indirect effect is slower than the direct effect.