• Molecules and Cells
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• v.44 no.2
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• pp.88-100
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• 2021

Anoctamin 6/TMEM16F (ANO6) is a dual-function protein with Ca2+-activated ion channel and Ca2+-activated phospholipid scramblase activities, requiring a high intracellular Ca2+ concentration (e.g., half-maximal effective Ca2+ concentration [EC50] of [Ca2+]i > 10 μM), and strong and sustained depolarization above 0 mV. Structural comparison with Anoctamin 1/TMEM16A (ANO1), a canonical Ca2+-activated chloride channel exhibiting higher Ca2+ sensitivity (EC50 of 1 μM) than ANO6, suggested that a homologous Ca2+-transferring site in the N-terminal domain (Nt) might be responsible for the differential Ca2+ sensitivity and kinetics of activation between ANO6 and ANO1. To elucidate the role of the putative Ca2+-transferring reservoir in the Nt (Nt-CaRes), we constructed an ANO6-1-6 chimera in which Nt-CaRes was replaced with the corresponding domain of ANO1. ANO6-1-6 showed higher sensitivity to Ca2+ than ANO6. However, neither the speed of activation nor the voltage-dependence differed between ANO6 and ANO6-1-6. Molecular dynamics simulation revealed a reduced Ca2+ interaction with Nt-CaRes in ANO6 than ANO6-1-6. Moreover, mutations on potentially Ca2+-interacting acidic amino acids in ANO6 Nt-CaRes resulted in reduced Ca2+ sensitivity, implying direct interactions of Ca2+ with these residues. Based on these results, we cautiously suggest that the net charge of Nt-CaRes is responsible for the difference in Ca2+ sensitivity between ANO1 and ANO6.

• Journal of dental hygiene science
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• v.2 no.1
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• pp.47-51
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• 2002

The concentration of intracellular $Ca^{2+}$ was measured by spectrofluorometer after rat submandibular acinar cells were loaded with fura-2/AM(fura-2). After isoproterenol and octanol were administered while letting submandibular gland acinar cell placed in a perfusion chamber flow through a standard solution, the changes of $Ca^{2+}$ concentration were measured. When they were administered separately, there showed little changes of intracellular $Ca^{2+}$ concentration. When they were administered at the same time, however the concentration of intracellar $Ca^{2+}$ was shown to increase. When forskolin, an adenylate cyclase activater, was administerd together with octanol the response looked similar to the response of isoproterenol. In case of the extracellular $Ca^{2+}$ was removed by omitting $Ca^{2+}$ in standard solution and treating EGTA, isoproterenol and forskolin does not affect to the concentration of intracellular $Ca^{2+}$. Therefore, it was certified that the increase of intracellular $Ca^{2+}$ was caused from outside the cell. In order to know that the $Ca^{2+}$ influx is related with capacitative entry pathway, godolinium, blocker of that pathway was treated. With the result of that experiment there was no complete control of the increase in the concentration of intracellular $Ca^{2+}$. However, speed and amount of $Ca^{2+}$ increase was comparatively diminished.

• Journal of Embryo Transfer
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• v.20 no.3
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• pp.191-200
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• 2005

Many studies have shown that the development of mouse early 2-cell embryos in vitro is related with the intracellular $Ca^{2+}$ changes. In ICR strain mouse, the development of embryos arrests at early 2-cell stage, but the arrested early 2-cell embryos can be rescued by the addition of $Ca^{2+}$-related materials. Acetylcholine (ACh) increases intracellular $Ca^{2+}$ concentration ([$Ca^{2+}$]i) via the mAChR-PLC-IP3 pathway in mouse oocytes. We examined whether ACh rescues 2-cell block in mouse. In early 2-cell embryos, ACh increased [$Ca^{2+}$]i in a dose-dependent manner (p<0.001), and had an effect on rescue of 2-cell block and embryonic development. To identify the signal pathway involved in ACh-induced rescue of 2-cell block, we first applied an agonist of ACh receptor (AChR). Like ACh, carbachol increased intracellular $Ca^{2+}$ concentration ([$Ca^{2+}$]i) and atropine, an antagonist of ACh receptor, blocked the ACh-induced $Ca^{2+}$ increase. In $Ca^{2+}$-free medium, ACh also increased [$Ca^{2+}$]i, indicating that $Ca^{2+}$ increased by ACh is mainly released from the intracellular $Ca^{2+}$ store. The ACh-induced $Ca^{2+}$ increase was blocked by PLC inhibitor (U73122), ryanodine receptor (RyR) antagonist (dantrolene), and CaM KII inhibitor (KN-93), but not by IP3R antagonists (xestospongin C). These results show that ACh increases intracellular $Ca^{2+}$ concentration via mAChR/PLC/RyR, and this contributes to the rescue of 2-cell block.

• Proceedings of the Korean Ceranic Society Conference
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• 2002.10a
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• pp.30.1-30
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• 2002

• The Journal of Korean Institute of Communications and Information Sciences
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• v.18 no.8
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• pp.1102-1107
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• 1993

Microwave dielectric properties of CaTiO3 and CaTiO3-TiO2 ceramics for the composition range between 40 and 50 mol% CaO in CaO-TiO2 binary system were investigated. CaTiO3 ceramics with50 mol% CaO showed the dielectric constant (e,) of 178, the temperature coefficient of resonant frequency(c,) of+1000 ppm/'c and the qualify factor Q of 2760 (f0=2.7 GHz ). Dielectric constant and temperature coefficient of resonant frequency of ceramics with dual phases of CaTiO3 and TiO2 decreased gradually from those of CaTiO3 as the CaO content decreased. Q value and density were found to have minimum at the composition of 47 mol% CaO. The degradation of Q value and density in dual phase ceramics seems to be caused by the large pores at grain boundaries and/or within grains remained after rapid growth of CaTiO3 grains as TiO2 Phase decreased.

• YAKHAK HOEJI
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• v.50 no.2
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• pp.111-117
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• 2006

Cardiac chambers serve as mechanosensory systems during the haemodynamic or mechanical disturbances. To examine a possible role of fluid pressure (FP) in the regulatien of atrial $Ca^{2+}$ signaling we investigated the effect of FP on L-type $Ca^{2+}$ current $(I_{Ca})$ in rat ventricular myocytes using whole-cell patch-clamp technique. FP $(\sim40cm\;H_2O)$ was applied to whole area of single myocytes with electronically controlled micro-jet system. FP suppressed the magnitude of peak $I_{Ca}$ by $\cong25\%$ at 0 mV without changing voltage dependence of the current-voltage relationship. FP significantly accelerated slow component in inactivation of $I_{Ca}$, but not its fast component. Analysis of steady-state inactivation curve revealed a reduction of the number of $Ca^{2+}$ channels available for activity in the presence of FP. Dialysis of myocytes with high concentration of immobile $Ca^{2+}$ buffer partially attenuated the FP-induced suppression of $I_{Ca}$. In addition, the intracellular $Ca^{2+}$ buttering abolished the FP-induced acceleration of slow component in $I_{Ca}$ inactivation. These results indicate that FP sup-presses $Ca^{2+}$ currents, in part, by increasing cytosolic $Ca^{2+}$ concentration.

• Journal of Bio-Environment Control
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• v.11 no.3
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• pp.127-132
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• 2002

Enhanced supply of $Ca^{2＋}$ as well as NO$_3$$^{[-10]}$ is known to restrict the uptake of the Na$^{＋}$ and Cl$^{[-10]}$ ion and ameliorate growth under saline conditions. This test was conducted to investigate the ameliorating effects of Ca(NO$_3$)$_2$ or CaCl$_2$ on the growth and yield of NaCl-stressed tomato plants grown in plastic pot filled with soil. All treatments except for the control were supplied with 80 mM NaCl fur two weeks after transporting. The saline solutions with nutrient were supplemented with either 0, 10 or 20 mM Ca(NO$_3$)$_2$ and either 0, 10 or 20 mM CaCl$_2$ during harvesting time from two weeks after transporting. Ca(NO$_3$)$_2$ or CaCl$_2$ application enhanced the growth such as plant height, fresh weight, dry weight, fruit number, and fruit weight, and yield of NaCl-stressed tomato, and also their effects increased greater as concentration of supplemented Ca(NO$_3$)$_2$ or CaCl$_2$increased. Yield increased in 20 mM Ca(NO$_3$)$_2$ compared with the others except fur the control. Photosynthetic rate in Ca treatments was lower than that of the control, but higher than that of NaCl treatment. Leaf chlorophyll content was higher in Ca treatments compared with the others, especially in younger leaf, while that was not affected by concentration of supplemented Ca. Ca(NO$_3$)$_2$ or CaCl$_2$ supply increased the $K^{＋}$ and $C^{2＋}$ concentration of tomato plants, whereas the Na$^{＋}$ transport to the leaves was inhibited. There was a strong increase in the $K^{＋}$/Na$^{＋}$ ratio in plants treated Ca(NO$_3$)$_2$, or CaCl$_2$. Cl$^{[-10]}$ content of plants was decreased by supplemental Ca(NO$_3$)$_2$ but Cl$^{[-10]}$ was increased in plants with CaCl$_2$compared with Ca(NO$_3$)$_2$. N concentration in plants of tomato increased with enhanced Ca(NO$_3$)$_2$ or CaCl$_2$supply, In conclusion, our study confirms the potential of Ca(NO$_3$)$_2$ or CaCl$_2$to alleviate NaCl-induced growth reductions in tomato.

• Journal of Microbiology
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• v.34 no.3
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• pp.253-269
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• 1996

Infection of fish cells with IHNV resulted in gradual increase in cytosolic free Ca$\^$2+/ concentration ([Ca$\^$2+/)] in CHSE, gradual decrease in [Ca$\^$2+/] in FHM, and no significant change in RTG cells. The degree of [Ca$\^$2+/] increase or decrease was dependent on the amount of infectious virus, and these [Ca$\^$2+/] variations were maximal at 16 hours after virus infection (p. i.) in both cell lines. When the fish cells were infected with inactivated IHNV, evident variation in [Ca$\^$2+/] was not observed. Thus, infectivity of IHNV appears to correlate with changes in [Ca$\^$2+/] in virus-infected cells. These IHNV-induced [Ca$\^$2+/] changes were partially blocked by cycloheximide, but not affected by cordycepin. It seems to be that virus-induced Ca$\^$2+/ variations were more related with protein synthesis than RNA synthesis. Various Ca$\^$2+/ related drugs were used in search for the mechanisms of the [Ca$\^$2+/], changes following IHNV infection of CHSE cells. Decreasing extracellular Ca$\^$2+/ concentration or blocking Ca$\^$2+/ influx from extracellular media inhibited the IHNV-induced increase in [Ca$\^$2+/], in CHSE cells. Similar results were obtained with intracellular Ca$\^$2+/ sources are important in IHNV-induced [Ca$\^$2+/] increase in CHSE cells.

• The Korean Journal of Physiology and Pharmacology
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• v.9 no.2
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• pp.95-101
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• 2005

In the heart, $Na^{+}-Ca^{2+}$ exchange (NCX) is the major $Ca^{2+}$ extrusion mechanism. NCX has been considered as a relaxation mechanism, as it reduces global $[Ca^{2+}]_i$ raised during activation. However, if NCX locates in the close proximity to the ryanodine receptor, then NCX would curtail $Ca^{2+}$ before its diffusion to global $Ca^{2+}_i$ This will result in a global $[Ca^{2+}]_i$ decrease especially during its ascending phase rather than descending phase. Therefore, NCX would decrease the myocardial contractility rather than inducing relaxation in the heart. This possibility was examined in this study by comparing NCX-induced extrusion of $Ca^{2+}$ after its release from SR in the presence and absence of global $Ca^{2+}_i$ transient in the isolated single rat ventricular myocytes by using patch-clamp technique in a whole-cell configuration. Global $Ca^{2+}_i$ transient was controlled by an internal dialysis with different concentrations of BAPTA added in the pipette. During stimulation with a ramp pulse from +100 mV to -100 mV for 200 ms, global $Ca^{2+}_i$ transient was suppressed only mildly, and completely at 1 mmol/L, and 10 mmol/L BAPTA, respectively. In these situations, ryanodine-sensitive inward NCX current was compared using $100{\mu}mol/L$ ryanodine, $Na^+$ depletion, 5 mmol/L $NaCl_2$ and $1{\mu}mol/L$ nifedipine. Surprisingly, the result showed that the ryanodine-sensitive inward NCX current was well preserved after 10 mmol/L BAPTA to 91 % of that obtained after 1 mmol/L BAPTA. From this result, it is concluded that most of the NCX-induced $Ca^{2+}$ extrusion occurs before the $Ca^{2+}$ diffuses to global $Ca^{2+})i$ in the rat ventricular myocyte.

• Analytical Science and Technology
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• v.24 no.6
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• pp.527-532
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• 2011

IQ motif-containing GTPase-activating protein 1 (IQGAP1), which is a well-known $Ca^{2+}$-independent calmodulin (CaM) binding protein, is involved in various cellular functions such as cell proliferation and cell migration. IQGAP1 has four repeated IQ motifs, which are crucial for CaM binding. It has been shown that all four IQ motifs of IQGAP1 can bind to $Ca^{2+}$/CaM, while the third and fourth IQ motifs of IQGAP1 can bind to apoCaM. However, it has not been clear whether the CaM binding of IQ motifs of IQGAP1 was mediated directly or indirectly. In this study, we examined whether the binding between CaM and each IQ motif of IQGAP1 was direct in vitro. As a result, we found that IQ1 motif has a weak $Ca^{2+}$-dependent CaM binding. In contrast, IQ3 has a $Ca^{2+}$-dependent CaM binding. All other motifs have no significant CaM binding. We also found that IQ(2.7-3) and IQ(3.5-4.4) have CaM binding capacity. This finding indicates that IQ motifs of IQGAP1 plays a dynamic role via different motif interactions with $Ca^{2+}$/CaM or proCaM.