• Title/Summary/Keyword: Calcium current

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Biochemical Characterization of a Putative Calcium Influx Factor as a Diffusible Messenger in Jurkat Cells, Xenopis Oocytes, and Yeast

  • Kim, Hak-Yong
    • Animal cells and systems
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    • v.7 no.1
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    • pp.75-79
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    • 2003
  • Highly purified high performance thin layer chromatography (HPTLC) fractions containing a putative calcium influx factor (CIF) were prepared from the Jurkat cells and Xenopus oocytes in which $Ca^{2+}$ stores were depleted by thapsigargin treatment and from the yeast in which intracellular $Ca^{2+}$ stores were also depleted by genetic means. Microinjection of the fractions has been shown to elicit $Ca^{2+}$ dependent currents in Xenopus oocytes. The nature of the membrane currents evoked by the putative CIF appeared to be carried by chloride ions since the current was blocked by the selective chloride channel blocker 1 mM niflumic acid and its reversal potential was about -24 mV. Injection of the calcium chelator 1,2-bis(2-aminophenoxy)ethane-N, N, N',N'-tetraacetic acid (BAPTA) eradicated the current activities, suggesting the current responses are entirely $Ca^2$-dependent. Moreover, the currents were sensitive to the removal of extracellular calcium, indicating the dependence on calcium entry through the plasma membrane calcium entry channels. CIF activities were insensitive to protease, heat, and acid treatments and to Dische-reaction whereas the activities were sensitive to nucleotide pyrophosphatase and hydrazynolysis. The fraction might have a sugar because it was sensitive to Molisch test and Seliwaniff's resorcinol reaction. From the above results, CIF as a small and stable molecule seems to have pyrimidine, pyrophosphate, and a sugar moiety.oiety.

The role of calmodulin in regulating calcium-permeable PKD2L1 channel activity

  • Park, Eunice Yon June;Baik, Julia Young;Kwak, Misun;So, Insuk
    • The Korean Journal of Physiology and Pharmacology
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    • v.23 no.3
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    • pp.219-227
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    • 2019
  • Polycystic kidney disease 2-like-1 (PKD2L1), polycystin-L or transient receptor potential polycystin 3 (TRPP3) is a TRP superfamily member. It is a calcium-permeable non-selective cation channel that regulates intracellular calcium concentration and thereby calcium signaling. Although the calmodulin (CaM) inhibitor, calmidazolium, is an activator of the PKD2L1 channel, the activating mechanism remains unclear. The purpose of this study is to clarify whether CaM takes part in the regulation of the PKD2L1 channel, and if so, how. With patch clamp techniques, we observed the current amplitudes of PKD2L1 significantly reduced when co-expressed with CaM and $CaM{\triangle}N$. This result suggests that the N-lobe of CaM carries a more crucial role in regulating PKD2L1 and guides us into our next question on the different functions of two lobes of CaM. We also identified the predicted CaM binding site, and generated deletion and truncation mutants. The mutants showed significant reduction in currents losing PKD2L1 current-voltage curve, suggesting that the C-terminal region from 590 to 600 is crucial for maintaining the functionality of the PKD2L1 channel. With PKD2L1608Stop mutant showing increased current amplitudes, we further examined the functional importance of EF-hand domain. Along with co-expression of CaM, ${\triangle}EF$-hand mutant also showed significant changes in current amplitudes and potentiation time. Our findings suggest that there is a constitutive inhibition of EF-hand and binding of CaM C-lobe on the channel in low calcium concentration. At higher calcium concentration, calcium ions occupy the N-lobe as well as the EF-hand domain, allowing the two to compete to bind to the channel.

Regulatory Action of $\beta-adrenergic$ Agonist and 8-bromocyclic AMP on Calcium Currents in the Unfertilized Mouse Eggs

  • Haan, Jae-Hee;Cheong, Seung-Jin;Kim, Yang-Mi;Park, Choon-Ok;Hong, Seong-Geun
    • The Korean Journal of Physiology
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    • v.27 no.2
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    • pp.175-183
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    • 1993
  • There are many report suggesting that influx and intracellular calcium concentration $([Ca^{2+}]_i)$ are related to cell signalling in various cells. However, it has not been reported that calcium channel activation is affected by the substances involved in signal transduction pathways in the mouse eggs. In this study, the effects of isoprenaline (ISP) and cyclic AMP on calcium influx through calcium channels were investigated to show their relationship with the signal transduction process in unfertilized mouse eggs. Using whole cell voltage clamp techniques, calcium currents, elicited by the depolarizing pulses of 300 ms duration (from -50 mV to 50 mV in 10 mV increments) from a holding potential of -80 mV, were recorded. The current-voltage (I-V) relation of calcium currents was shown to be bell-shaped; the current began to activate at -50 mV and reached its maximum $(-1.33{\pm}0.16\;nA:\;mean{\pm}S.E.,\;n=7)$ at -10 mV, then decayed at around 50 mV. Calcium currents were fully activated within $7\;ms{\sim}20\;ms$ and completely inactivated 200 ms after onset of the step pulse. ISP within the concentration ranges of $10^{-8}\;M{\sim}10^{-4}\;M$ dose-dependently increased the amplitude calcium current. The permeable cyclic AMP analogue,8-bromocyclic AMP, also increased its maximal amplitude by 46ft at $10^{-5}\;M$, while protein kinase inhibitor (PKI), which is known to inhibit 0.02 phosphorylating units of cyclic AMP-dependent protein kinase (PKA) per microgram decreased calcium currents. Currents recorded in the presence of PKI were resistant to increase by the application of $10^{-5}\;M$. Also, PKI inhibited the calcium current increase elicited by ISP treatment. These results suggest that $\beta-adrenergic$ regulation of the calcium channel is mediated by the cAMP-dependent protein kinase. This signal transduction pathway might play a role in regulating $[Ca^{2+}]_i$, level due to the increase of calcium influx in mouse eggs.

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Influence of Ca-Si Addition on Anodic Polarization Chgaraqcteristics of Al-Zn-In Anodes (海水中 Al-Zn-In 合金陽極의 分極特性에 미치는 Ca-Si 添加의 影響)

  • Seo, Chang-Je
    • Journal of the Korean institute of surface engineering
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    • v.12 no.1
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    • pp.3-10
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    • 1979
  • Many excellent Al-Zn-In anode have been developed up to the present. But for the purpose of the better performance of Al-Zn-In anodes in sea water the effect of calcium silicon addition on anodic polarization and current capacity of Al-Zn-In anodes was measured and analysed in sea water and artificial sea water. The results and conclusions obtained are summarized as follows. 1) Being compared with Al-Zn-In anodes, Al-Zn-In anodes containing 0.05% calcium silicon had superior characteristics in both anodic polarization and current capacity. 2) Corrosion patterns of the anodes containing calcium silicon were much more uniform than those of Al-Zn-In anodes. 3) In this experiment the most useful anode was Al-4% Zn-0.03% In-0.05% (Ca-Si). It had a capacity of 2.60Amp-hr of current/g and a voltage of 1.13(SCE reference) at anodic current density 1,000 4{\mu}A/cm^2$.

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Calcium Homeostasis and Regulation of Calbindin-D9k by Glucocorticoids and Vitamin D as Bioactive Molecules

  • Choi, Kyung-Chul;Jeung, Eui-Bae
    • Biomolecules & Therapeutics
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    • v.17 no.2
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    • pp.125-132
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    • 2009
  • Calbindin-$D_{9k}$ (CaBP-9k), a cytosolic calcium-binding protein, is expressed in a variety of tissues, i.e., the duodenum, uterus, placenta, kidney and pituitary gland. Duodenal CaBP-9k is involved in intestinal calcium absorption, and is regulated at transcriptional and post-transcriptional levels by 1,25-dihydroxyvitamin D3, the hormonal form of vitamin D, and glucocorticoids (GCs). Uterine CaBP-9k has been implicated in the regulation of myometrial action(s) through modulation of intracellular calcium, and steroid hormones appear to be the main regulators in its uterine and placental regulation. Because phenotypes of CaBP-9k-null mice appear to be normal, other calcium-transporter genes may compensate for its gene deletion and physiological function in knockout mice. Previous studies indicate that CaBP-9k may be controlled in a tissue-specific fashion. In this review, we summarize the current information on calcium homeostasis related to CaBP-9k gene regulation by GCs, vitamin D and its receptors, and its molecular regulatory mechanism. In addition, we present related data from our current research.

The Characterization of the Increase of Membrane Conductance after Depolarization in Single Rat Adrenal Chromaffin Cells

  • Lim, Won-Il;Kim, Sang-Jeong;Kim, Jun
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.1
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    • pp.95-100
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    • 1998
  • The conductance change evoked by step depolarization was studied in primarily cultured rat adrenal chromaffin cells using patch-clamp and capacitance measurement techniques. When we applied a depolarizing pulse to a chromaffin cell, the inward calcium current was followed by an outward current and depolarization-induced exocytosis was accompanied by an increase in conductance trace. The slow inward tail current which has the same time course as the conductance change was observed in current recording. The activation of slow tail current was calcium-dependent. Reversal potentials agreed with Nernst equation assuming relative permeability of $Cs^+\;to\;K^+$ is 0.095. The outward current and tail current were blocked by apamin (200 nM) and d-tubocurarine (2 mM). The conductance change was blocked by apamin and did not affect membrane capacitance recording. We confirmed that conductance change after depolarization comes from the activation of the SK channel and can be blocked by application of the SK channel blockers. Consequently, it is necessary to consider blocking of the SK channel during membrane capacitance recording.

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Two Types of Voltage-activated Calcium Currents in Goldfish Horizontal Cells

  • Paik, Sun-Sook;Bai, Sun-Ho;Jung, Chang-Sub
    • The Korean Journal of Physiology and Pharmacology
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    • v.9 no.5
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    • pp.269-273
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    • 2005
  • In horizontal cells (HCs) that were freshly dissociated from goldfish retina, two types of voltagedependent calcium currents ($I_{Ca}$) were recorded using a patch-clamping configuration: a transient type current and a sustained type current. The cell was held at -40 mV, and the prepulse step of -90 mV was applied before command pulse between -65 and +55 mV. The transient $Ca^{2+}$ current was activated by depolarization to around -50 mV from a prepulse voltage of -90 mV lasting at least 400 ms and reached a maximal value near -25 mV. On the other hand, the sustained $Ca^{2+}$ current was induced by pre-inactivation for less than 10 ms duration. Its activation started near -10 mV and peaked at +20 mV. $Co^{2+}$ (2 mM) suppressed both of these two components, but nifedipine ($20{\mu}M$), L-type $Ca^{2+}$ channel antagonist, blocked only the sustained current. Based on the activation voltage and the pharmacolog$I_{Ca}$l specificity, the sustained current appears to be similar to L-type $I_{Ca}$ and the transient type to T-type $I_{Ca}$. This study is the first to confirm that transient type $I_{Ca}$ together with the sustained one is present in HCs dissociated from goldfish retina.

Effect of milk and milk products consumption on physical growth and bone mineral density in Korean adolescents

  • Kim, Sun Hyo;Kim, Woo Kyoung;Kang, Myung-Hee
    • Nutrition Research and Practice
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    • v.7 no.4
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    • pp.309-314
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    • 2013
  • This study was conducted to investigate the relationship among the current status of calcium intake from milk and milk products, physical growth and bone mineral density in 664 male and female middle school and high school students aged 15-17 years. In the study, the current status of calcium intake from milk and milk products was analyzed, and the height, body composition, and bone mineral density of the right heel bone (calcaneus) were measured. The daily calcium intake of milk and milk products was calculated as the 'dairy equivalent of calcium', which is the calcium content in 200 mL of white milk. The cutoffs of tertiles of the dairy equivalent of calcium were calculated and then the subjects were categorized into 3 groups according to the tertiles, Q1 group (lower intake group), Q2 group (middle intake group) and Q3 group (upper intake group). The daily calcium intake of milk and milk products in Q1, Q2 and Q3 groups was 16.2 mg, 99.7 mg, and 284.0 mg, respectively, and the ratio of milk and milk product consumption to the daily total calcium intake was 5.4%, 27.4%, and 49.7%, respectively. The ratio of total calcium intake to the daily recommended intake in study subjects was 30.5% in Q1, 42.3% in Q2, and 60.7% in Q3, with significant differences (P < 0.05). Height, body weight, BMI, and % of body fat in three tertile groups (Q1, Q2 and Q3) were not significantly different. However, the T scores for bone mineral density in female students in three tertile groups (Q1, Q2 and Q3) was significantly different (P < 0.05). The study showed that the intake of milk and milk products in adolescents, particularly in girls, can improve the bone mineral density without increasing body weight, and thus confirmed that milk intake is important in adolescence.

[$Cl^-$-sensitive Component of $Ca^{2+}$-activated Tail Current in Rabbit Atrial Myocytes

  • Park, Choon-Ok;So, In-Suk;Ho, Won-Kyung;Kim, Woo-Gyeum;Earm, Yung-E
    • The Korean Journal of Physiology
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    • v.26 no.1
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    • pp.27-35
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    • 1992
  • We used the whole cell patch clamp technique to examine the ionic basis for the tail current after depolarizing pulse in single atrial myocytes of the rabbit. We recorded the tail currents during various repolarizations after short depolarizing pulse from a holding potential of -70 mV. The potassium currents were blocked by external 4-aminopyridine and replacement of internal potassium with cesium. The current was reversed to the outward direction above +10 mV. High concentrations of intracellular calcium buffer inhibited the activation of the current. Diltiazem and ryanodine blocked it too. These data suggest that the current is activated by intracellular calcium released from sarcoplasmic reticulumn. When the internal chloride concentration was increased, the inward tail current was increased. The current was partially blocked by the anion transport blocker niflumic acid. The current voltage curve of the niflumic acid sensitive current component shows outward rectification and is well fitted to the current voltage curve of the theoretically predicted chloride current calculated from the constant field equation. The currents recorded in rabbit atrial myocytes, with the method showing isolated outward Na Ca exchange current in ventricular cells of the guinea pig, suggested that chloride conductance could be activated with the activation of Na/ca exchange current. From the above results it is concluded that a chloride sensitive component which is activated by intracellular calcium contributes to tail currents in rabbit atrial cells.

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Current status in calcium biofortification of crops (작물의 생합성 칼슘 함량 증대 연구 현황)

  • Lee, Jeong-Yeo;Nou, Ill-Sup;Kim, Hye-Ran
    • Journal of Plant Biotechnology
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    • v.39 no.1
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    • pp.23-32
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    • 2012
  • Calcium is an essential nutrient for living organisms, with key structural and signaling roles. Its deficiency in plants can result in poor biotic and abiotic stress tolerance as well as reduced crop quality and yield. Calcium deficiency in humans causes various diseases such as osteoporosis and rickets. Biofortification of calcium in various food crops has been suggested as an economic and environmentally advantageous method to enhance human intake of calcium. Recent efforts to increase the levels of calcium in food crops have used calcium/proton antiporters ($CAXs$) and modified one to increase calcium transport into vacuoles through genetic engineering. It has been reported that overall calcium content of transgenic plants has been increased in their edible portions with some adverse effects. In conclusion, biofortification of calcium will add more value in crops as well as will be beneficial for animal and human. Therefore, more fundamental studies on the mechanisms of calcium ion storage and transporting are essential for more effective calcium biofortification.