• Title/Summary/Keyword: $Ca^{++}$uptake

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Scorpion Venom Activates Both $Ca^{2+}-ATPase$ and Inositol 1,4,5-trisphosphate Receptor in the Microsomes of Tracheal Epithelial Cells (전갈독소에 의한 호흡기 상피세포 마이크로솜 $Ca^{2+}-ATPase$와 Inositol 1,4,5-trisphosphate 수용체의 활성촉진)

  • Cho, Kyong-Soo;Park, Kyoung-Sun;Kim, Young-Kee
    • Applied Biological Chemistry
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    • v.39 no.3
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    • pp.189-194
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    • 1996
  • The effects of scorpion (Leiurus quinquestriatus hebraeus, Lqh) venom were evaluated on the activities of microsomal $Ca^{2+}-ATPase$ and $Ca^{2+}$ release channel prepared from the epithelial cells of pig airway. Whole venom of Lqh $(120\;{\mu}g/ml)$ increased the activity of microsomal $Ca^{2+}-ATPase$ about 32% in the tight-sealed microsomes and about 28% in the Triton X-100-treated or $Ca^{2+}$ ionophore A23187-treated leaky microsomes. Thapsigargin, a specific antagonist of $Ca^{2+}-ATPase$, inhibited 42% of total ATPase activity and also completely blocked the effects of Lqh venom, suggesting that Lqh venom directly activiates the microsomal $Ca^{2+}-ATPase$. In order to determine if Lqh venom increases the microsomal uptake of $^{45}Ca^{2+}$, Lqh venom was added in the uptake medium. The Lqh venom increased microsomal $^{45}Ca^{2+}$ uptake up to ${\sim}20%$ and the increase was only observed when heparin, an antagonist of $InsP_3$ receptor channel, was added in the uptake medium. Lqh venom in the absence of heparin unexpectedly decreased the rate and the amount of $^{45}Ca^{2+}$ uptake. These results were explained by simultaneous increases in $^{45}Ca^{2+}$ release as well as $^{45}Ca^{2+}$ uptake by Lqh venom. Lqh venom itself increased the release of $^{45}Ca^{2+}$ as much as $^{45}Ca^{2+}$ release by $4\;{\mu}m\;InsP_3$, implying that Lqh venom also activates $InsP_3$ receptor, microsomal $Ca^{2+}$ release channel. Based on these results, we suggest that the Lqh venom consists of at least two components; one activates the $InsP_3$ receptor and the other avates the $Ca^{2+}-ATPase$. Currently we a investigating the chemical and electrophysiological properties of the active components of Lqh venom.

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The Effects of Green Tea Catechins on Vascular Smooth Muscle Tension and 45 $Ca^{2+}$ Uptake (녹차 카테킨류의 혈관장력 및 $Ca^{2+}$유입에 미치는 영향)

  • 안희열;이미애;윤여표
    • Journal of Food Hygiene and Safety
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    • v.11 no.2
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    • pp.83-87
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    • 1996
  • The objective of this study is to investigate the direct effects of green tea catechins(GTC) on vascular smooth muscle tension and 45Ca2+ Uptake in rat aorta. The methods used in this study are isometric tension measurements using physiograph, Lanthanum method for 45Ca2+(2 uCi/ml) uptake measurement in rat aorta. GTC modified tension induced by 40 mM KCl or 1 uM norepinephrine in rat aorta. Low concentrations of GTC(<0.5mg/ml) increased tension by 40 mM KCl or 1 uM norepinephrine, individually. However, high conecentration of GTC(>0.5 mg/ml) inhibiited tension by 40 mM KCl or 1 uM norepinephrine, individually. GTC increased 45Ca uptake induced by 40 mM KCl in a dose-dependent manner. From these results, GTC has the dual actions in vascular smooth muscle in vitro. Low concentrations of GTC augments tension by K or norepinephrine. However, high concentrations of GTC inhibits tension by K or norepinephrine GTC may have Ca2+ channel activation, action, which may result in unphysiological vasodilation by Ca2+ overload in vascular smooth muscle.

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Increase in $Na^+-Ca^{2+}$ Exchange Activity in Sarcolemma Isolated from Mesenteric Arteries of Spontaneously Hypertensive Rats

  • Lee, Shin-Woong;Lee, Jeung-Soo;Park, Young-Joo;Park, In-Sook
    • Archives of Pharmacal Research
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    • v.12 no.2
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    • pp.128-134
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    • 1989
  • $Na^+-Ca^{2+}$ exchange process in sarcolemmal vesicles isolated from mesenteric arteries of Wistar-Kyoto normotensive(WKY) and spontaneously hypertensive rats(SHR) was investigated. The sarcolemmal fractions isolated after homogenization and sucrose density gradient centrifugation were enriched with 5'-nucleotidase and ouabain sensitive, $K^+-dependent$ phosphatase activities. When the vesicles were loaded with $Na^+$, a time dependent $Ca^{2+}$ uptake was observed. However, very little $Ca^{2+}$ uptake was observed when the vesicles were loaded with $K^+$, or $Ca^{2+}$ uptake of the $Na^+-loaded$ vesicles was carried out in high sodium medium so that there was no sodium gradient. When the vesicles loaded with $Ca^{2+}$ by $Na^+-Ca^{2+}$ exchange were diluted into potassium medium containing EGTA, $Ca^{2+}$ was rapidly released from the vesicles. $Na^+-dependent\;Ca^{2+}$ uptake was increased in SHR compared to WKY, but passive efflux of preaccumulated $Ca^{2+}$ from the vesicles was decreased in SHR. The data indicate that the membrane vesicles of rat mesenteric arteries exhibit $Na^+-Ca^{2+}$ exchange activity. It is also suggested that changes of this process in vascular smooth muscle cell membrane of SHR may be involved in higher intracellular $Ca^{2+}$ concentration and higher basal tone in SHR.

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$[^3H]$ Ouabain Binding and Effect of Ouabain on $^{45}Ca^{2+}$-Uptake in Rat Cardiac Myocytes (쥐 심근 세포의 $[^3H]$ Ouabain 결합과 $^{45}Ca^{2+}}$섭취에 미치는 Ouabain의 영향)

  • 이신웅;김영희;진갑덕
    • YAKHAK HOEJI
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    • v.28 no.3
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    • pp.129-138
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    • 1984
  • Specific [$^{3}H$] ouabain binding and $Ca^{2+}$ -uptake were measured to elucidate the role of high affinity [$^{3}H$] ouabain binding site in rat cardiac myocytes which contain 65% of rod cells. High affinity [$^{3}$H] ouabain binding site, which is about 3% of total pump sites, with apparent dissociation constant ($K_{D}$) of $1.1{\times}10^{-7}M$ and maximum binding site concentration (Bmax) of 1.2 pmol/mg protein ($1.754{\times}10^{5}cells$) were identified. At the concentration of $10^{-7}M$ to $10^{-4}M$, ouabain produced concentration dependent increase in $Ca^{2+}$-uptake of myocytes. The effect of ouabain on $Ca^{2+}$-uptake was not effected by membrane depolarization (elevated K+ in incubation medium) or verapamil. These results suggest that in rat ventricular myocytes the ouabain receptor complex to high affinity site may increase Na+ - $Ca^{2+}$ exchange across the sarcolemmal membrane by inhibition of Na+, K+ - ATPase.

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Studies on the ATPase Activity and Calcium Transport of Fragmented Sarcoplasmic Reticulum (膜 ATPase 活性과 Ca 透過性에 관한 硏究)

  • Ha, Doo-Bong
    • The Korean Journal of Zoology
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    • v.20 no.2
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    • pp.101-107
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    • 1977
  • The effects of sodium azide, cAMP, G-strophanthin and dicumarol on the ATP-ase activity and Ca uptake of the fragmented sarcoplasmic reticulum of skeletal muscle were studied and the effects were compared with respect to the enzymatic activity and Ca transport. Sodium azide (0.05 mM) and G-strophanthin (0.25mM) caused no inhibition on either ATPase activity or Ca uptake. cAMP($1\\times10^{-6}\\sim5\\times10^{-4}$) had no effect on ATPase activity while inhibited Ca uptake. Dicumarol (0.05 mM) did not inhibit ATPase activity but caused a decreased Ca uptake of heavier fraction (8,000-12,000xG) of the reticulum fragments.

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The Effect of $Ca^{2+}$ and Its Specific Time in the Maturation of Mammalian Oocyte (포유류의 난자성숙과 성숙과정에서 $Ca^{2+}$의 영향과 그 작용시기)

  • Bae, In-Ha
    • Clinical and Experimental Reproductive Medicine
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    • v.21 no.3
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    • pp.285-296
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    • 1994
  • Follicular oocytes were released from the graafian follicles of ovaries from 3-4 weeks old mice. The spontaenous maturation of these follicular oocyes was inhibited by the treatment of dbcAMP and progesterone and these oocytes were cultured for 2-8hr in the Modified Hank's balanced salt solution(MHBS). Ethylenediaminetetraaceticacid(EDTA) and calmoudulin antagonist, trifluoperazine (TFP) were treated to the culture medium in order to investigate whether these chemical agents inhibit calcium uptake into the oocyte and oocyte maturation. $^{45}Ca^{2+}$, 10-${\mu}$Ci/ml was added to the culture medium during the culture period. $^{45}Ca^{2+}$uptake into the oocytes was examined whether and when various kind of oocyte maturation inhibiting agents inhibit or stimulate the influx of calcium into oocytes. Dibutyryl cAMP and progesterone decrease $^{45}Ca^{2+}$uptake into the oocytes and synergistic inhibiting effect of dbcAMP and progesterone was prominent at much lower dosages. Calcium uptake into oocytes seems to be higher during first 2 hour culture period rather than next 4hr culture. After 8hr culture, calcium uptake level of the oocytes which GVBD already took place gradually approached to the level of those which were maintained at GV by the treatment of dbcAMP and progesterone. However, $^{45}Ca^{2+}$uptake into the GV maintained oocytes did not change at all even after 8hr culture period. In addition, calcium chelating agent, EDTA inhibited calcium uptake into oocytes as well as nuclear maturation of oocytes. Lower dosage used in the present study did not inhibit calcium uptake as well as oocyte maturation.

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Interactions between Estradiol-17 ${\beta}-BSA$ and Calcitropic Hormones in $Ca^{2+}$ Uptake in Renal Proximal Tubule Cells

  • Han, Ho-Jae;Lee, Yeun-Hee;Seo, Eun-Ju
    • The Korean Journal of Physiology and Pharmacology
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    • v.6 no.5
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    • pp.261-267
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    • 2002
  • The aim of the present study was to investigate the interaction of $estradiol-17{\beta}-bovine$ serum albumin $(E_2-BSA)$ and calcitropic hormones, such as parathyroid hormone, calcitonin, and vitamin D, in regulation of $Ca^{2+}$ uptake in primary cultured renal proximal tubule cells. Statistically significant increase in $Ca^{2+}$ uptake was found from 2 hours after $(E_2-BSA)\;(10^{-9}\;M)$ treatment, while $estradiol-17{\beta}\;(10^{-9}\;M)$ did not affect. Treatment of the cells with $(E_2-BSA)\;(10^{-9}\;M)$ together with parathyroid hormone (PTH) $(10^{-8}\;M),$ vitamin D $(10^{-8}\;M),$ or calcitonin $(10^{-8}\;M)$ significantly stimulated $Ca^{2+}$ uptake by 32.50%, 29.30%, or 27.75%, respectively, compared with the control. However, calcitropic hormones did not exhibit any synergistic effect on the E2-BSA-induced stimulation. $E_2-BSA$ significantly increased cAMP generation and PKC activity. The stimulatory effect of cotreatment of $E_2-BSA$ and PTH or vitamin D was blocked by SQ22536 (an adenylate cyclase inhibitor) and staurosporine (a PKC inhibitor), but the effect of cotreatment of $E_2-BSA$ and calcitonin was not blocked. Furthermore, 8-Br-cAMP and TPA (an artificial PKC promoter) increased $Ca^{2+}$ uptake by 25.51% and 16.47%, respectively, compared with the control. In conclusion, $E_2-BSA$ combined with calcitropic hormones regulated $Ca^{2+}$ uptake partially via cAMP and PKC-dependent mechanisms in renal proximal tubule cells.

Effect of Gonadotropin on $Ca^{++}$ Uptake in Follicle-Enclosed Mouse Oocytes Cultured in Vitro (배양된 생쥐여포에서 $Ca^{++}$ Uptake에 대한 Gonadotropin의 영향)

  • Bae, In-Ha;Kang, Shin-Hae
    • Clinical and Experimental Reproductive Medicine
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    • v.18 no.2
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    • pp.153-162
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    • 1991
  • The present study was undertaken to clarify the role of calcium ion as a factor for the maturation of follicle-enclosed mouse oocytes. Follicles were isolated with two sharp needles under a stereomicroscope from mouse(ICR) ovaries which were treated PMSG 5 IU 45 hours previously. Isolated follicles were cultured for 14-16 hours in an organ culture system at $37^{\circ}C$, 5% $CO_2$ in air and in a 100% humidified incubator by treatment of hCG, EDTA and $^{45}Ca^{++}$. Culture medium was Modified Hank's Balanced Salt Sol. (MHBS) and addition of hCG (human chorionic gonadotropin) was made into two doses level 0.4 IU and 0.8IU from the stock sol. and also $^{45}Ca^{++}$ was treated in the culture medium. To explain the role of calcium, calcium chelating agent EDTA was treated to the culture of the mouse follicle-enclosed oocytes. Two observations were made in the present study; nucleus phase and $^{45}Ca^{++}$ uptake into the oocyte. HCG induced oocyte maturation in the follicle about two folds as much as the control group, whereas there is no difference in oocyte maturation between 0.4 IU and 0.8 IU of hCG. Optimum level of hCG seems to be 0.4 IU/ml in the mouse follicle culture. HCG stimulated $^{45}Ca^{++}$ uptake into the oocyte of the follicles by two folds. $^{45}Ca^{++}$ uptake in the control group is about 2.5 folds in comparison of the EDTA(1.71mM) treated group. However, calcium uptake in the EDTA treated groups tends to increase depending on the decrease of EDTA concentration. These observations suggest that firstly, hCG stimulates maturation of the oocyte of the follicle, secondly, $Ca^{++}$ influx is induced by hCG and thirdly, $Ca^{++}$ influx by the treatment of EDTA decreases as a dosage-dependent process. This $Ca^{++}$ uptake may take place by the changes of permeability which was induced by hCG treatment. That is, $Ca^{++}$ influx may trigger the resumption of oocyte maturation. It is further necessary in the future study how this $Ca^{++}$ uptake is induced by hCG and increases permeability of the follicle and oocyte.

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The Effect of Ginseng on Heart Contraction and Sarcoplasmic Reticulum Function(II) The Effect of Ginseng on $^{45}Ca^{2+}$ Uptake by Sarcoplasmic Reticulum Fragments of Rat Heart

  • Sung, Baek-Yeon;Kim, Nak-Doo
    • Archives of Pharmacal Research
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    • v.6 no.1
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    • pp.69-73
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    • 1983
  • It was reported from our laboratory that the rate of deterioration of the force of contraction was slower in heart from Panax ginseng extract treated rats. Present investigation was designed to elucidate the mechanism of the slow deterioration of contractility of ginseng treated hearts. Therefore, $^{45}Ca^{2+}$ Uptake by sarcoplasmic reticulum (SR) isolated from ginseng treated rate and control rats was studied. Rate weighing 150-250g were administered orally with ginseng ethanol extract (100mg/kg) for 10 days. Cardiac SR was isolated by differential centrifugation and $^{45}Ca^{2+}$ uptake was assessed by the Millipore method. Freshly isolated SR from treated as well as control animals did not show any differences, but after incubation for 30 and 60 min at 37.deg.C, $^{45}Ca^{2+}$ uptake of control animal SR was found to be greatly depressed. The SR of treated animal possessed a greater degree of resistance to incubation. Thus it can be concluded that ginseng may have an ability to sustain the normal function of the heart by sustaining Ca accumulation by SR involved with the excitationcontraction coupling processes.

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Characteristics of Salt Adsorption by Calcium Alginate Beads (칼슘알긴산비드에 의한 염분의 흡착특성)

  • 방병호;서정숙
    • The Korean Journal of Food And Nutrition
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    • v.15 no.2
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    • pp.89-96
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    • 2002
  • The adsorption characteristics of sodium chloride into Ca-alginate beads have been investigated and the result were as follows: Sodium chloride uptake by Ca-alginate beads increased with time. The highest uptake volume of sodium chloride was 4.2g after 10 minutes. The uptake volume by Fe, Ca, Ba, and Sr-alginate beads was 5.6g, 4.2g, 4.2g and 4.0g, respectively but in case of Fe-alginate beads, the induced hydrogel beads were very fragile and the strength of Fe-alginate beads were weaker than Ca- and Ba-alginate beads. Mg-alginate bead was not formed and Ca-, Ba- and Sr-alginate beads had a similar uptake volume about 4.2g, respectively. The uptake volume of sodium chloride by CaCl$_2$concentration(0.1M. 0.2M and 1M), curing solution, was 4.8, 4.2g and 4.1g, respectively. The uptake volume by sodium alginate concentration(0.6%, 1% and 2%) was 2.8g, 4.0g, and 4.4g, respectively and Ca-alginate bead size was not effected in uptake sodium chloride. The uptake rate on initial sodium chloride concentration(4%, 8%, 12% and 16%) was 30%, 28%, 27% and 25%, respectively. The uptake rate on basic pH(10.0) was higher than when compared to other neutral pH(6.8) and acidic pH(4.0). The initial uptake velocity of sodium chloride from immobilization beads with salt resistant bacteria was lower than that of non-immobilization beads. The uptake rate of sodium chloride was decreased according to elongation of curing time. Reusability of Ca-alginate beads was possible but according to reutilization, the salt uptake volume of beads was also decreased. The uptake volume of sodium chloride from Doengjang by Ca-alginate beads on time course(3, 6, 12, and 24 hour) was revealed 5g, 6g, 7g and 7g, respectively.