• Title/Summary/Keyword: calcium ionophore

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Calcium Signaling of M II Oocyte during Chemical Activation of Calcium Ionopore and Cytochalasin B

  • Kim, Sung-Woo;Park, Jin-Ki;Park, Chun-Gyu;Lee, Ju-Young;Han, Joo-hee;Lee, Seung-Eun;Baek, Kyung-Nye;Chang, Won-Kyung
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.260-260
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    • 2004
  • The calcium ionophore (A23187) has been used for activation of porcine oocytes from in vitro maturation by many researches. The signaling of calcium was known to be a primary factor of activation of MII oocyte by calcium ionophore. The calcium level was measured by an intensity of fluo 4 fluorescence and confocal microscope. The level was increased by 7% ethanol or 70 μM calcium ionophore but oscillation was not found. (omitted)

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Concanavalin A Mediated Calcium Changes on Expansion and Hatching of the Mouse Blastocyst (Concanavalin A를 매개로한 세포내 Calcium의 변화와 생쥐 포배의 팽창과 부화)

  • Cheon, Yong Pil
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.4
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    • pp.337-346
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    • 2005
  • Objective: The oligosaccharide moieties of glycoproteins and proteoglycans have a vital function in blastocyst differentiation. Concanavalin (ConA), a lectin, is known to bind on the preimplantation embryos, especially on blastocyst. In this study, we investigated whether ConA can modulate the trophoblast development and about the regulating mediator. Also, we investigated whether expansion is enough for hatching procession of the mouse blastocyst. Method: Embryos were collected at 72 h post hCG injection and chemicals were treated after 24 h (96 hr post hCG injection). ConA or calcium ionophore A23187 were exposed to blastocyst and than analysis the developmental process for 48 hr. Intracellular free-$Ca^{2+}$ concentration in trophectoderm was measured with confocal laser microscope after exposing to ConA or calcium ionophore A23187. ConA-pretreated blastocyst exposed to the calcium ionophore A23187 and then analyzed the developmental process. Otherwise ouabain was treated to the blastocyst to block the $Na^+/K^+$-ATPase activity. Results: In contrast to the control blastocyst, the ConA-exposed blastocysts developed beyond the expansion stage with significantly high rate (90.4%) at 12 h post administration. ConA induced an increase the intracellular $Ca^{2+}$ concentration in trophectoderm. Calcium ionophore A23187 also stimulated expansion of blastocyst. Most of the control blastocysts developed to the hatching stage at 144 h post hCG injection. However, strongly 65% of the ConA-exposed embryos were arrested at expanded stage at same time point. The developmental progression rates to hatching stage of both ConA- and calcium ionophore A23187-expose blastocysts were significantly lower than that of the control. However ConA-pretreated embryos developed to the hatching stage like control embryos. Ouabain showed a tendency to delayed the progress to expansion stage but did not inhibit the development to the hatching stage. Conclusion: ConA-mediated expansion is the result of the increase of intracellular free-calcium in blastocyst stage embryo. It is suspected that expansion of the blasocyst is a essential indirect factor in hatching and the calcium may triggering the cellular mechanisms for the both expansion and hatching progression.

The effect of calcium ion concentrations in the medium and the treatment of caffeine and Ca-ionophore A23187 on in vitro capacitation of bull spermatozoa (배양액중의 calcium 이온 농도 및 caffeine과 Ca-ionophore A23187 처리가 소정자의 수정능획득에 미치는 영향)

  • Kim, Kye-Seung;Jo, Choong-Ho;Hwang, Woo-Seok
    • Korean Journal of Veterinary Research
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    • v.31 no.1
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    • pp.123-130
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    • 1991
  • The present study was performed to investigate the effect of Ca ion concentration on sperm viability and acrosome reaction rate and to evaluate the effect of treatments using caffeine and Ca-ionophore A23187 on acrosome reaction rate in frozen-thawed bull spermatozoa. Viabilities of in vitro capacitated bull spermatozoa at 0, 2.25 and 4.5 mM Ca ion concenrations were 21.00, 26.00 and 22.59%, respectively and significantly higher in Ca ion 2.25mM added group than Ca ion free group (p<0.05) and acrosome reaction rates of in vitro capacitated bull spermatozoa were 17.09, 52.15 and 47.92%, respectively and significantly high in Ca ion added groups(p<0.05). Viabilities in vitro capacitation by caffeine and Ca-ionophore A23187 in control, caffeine treated group, Ca-ionophore A23187 treated group and caffeine+Ca-ionophore A23187 treated group were 37.91, 27.67, 22.33 and 25.59%, respectively and significantly higher in control than treated groups(p<0.05), there were no significant differences among the treated groups, and acrosome reaction rates were 10.33, 37.92, 48.09 and 57.17%, respectively and there were significant differences among the groups(p<0.05), especially higher in caffeine+Ca-ionophore A23187 treated group than others.

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Effects of 1,25 Dihydroxycholecalciferol and Ca Ionophore A23187 on Ca Transports in Bone and Bone Cells (뼈조직과 세포에서의 칼슘이동기전에 대한 1,25 dihydroxycholecaliciferol과 Ca Ionophore A23187의 영향)

  • 이선영
    • Journal of Nutrition and Health
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    • v.21 no.3
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    • pp.173-181
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    • 1988
  • Various types of evidence suggest that some changes in cellular in cellular calcium may well signal the initiation of a chain of events leading to the physiological effects of the bone resorbing agents. The effects of 1,25-dihydorxycholecalciferol, $1.25\textrm{(OH)}_2\textrm{D}_3$, Ca ionophore A23187 and calcium antagonist, diltiazem on bone resprption and the cellular transport of Ca were investigated. Bone $^{45}\textrm{Ca}$ desaturation experiment was realized in isolated heterogenous rat bone cells after equilibrating the cells with $^{45}\textrm{Ca}$. Results of $^{45}\textrm{Ca}$ desaturation experiments were analysed by fitting the $^{45}\textrm{Ca}$ desaturation curve to a model of 2 exponential terms which indicated the presence of 2 exchangeable cellular calcium pools. $1.25\textrm{(OH)}_2\textrm{D}_3$ (0.5ng/$m\ell$) induced significantly bone resorption which was decreased by the physiological dose of diltiazeme(above 5nmol/$m\ell$) although it was ineffective alone. Ionophore A23187 (0.2$\mu\textrm{g}$/$m\ell$) decreased Ca release from bone but no additivity of effect with diltiazem(20nmol/$m\ell$) was observed. $1.25\textrm{(OH)}_2\textrm{D}_3$ (0.5ng/$10^{6}$ cells) had a moderate effect on the two kinetic phases of $^{45}\textrm{Ca}$ desaturation curve and these values were normalized when diltiazeme (20nmol/$10^{6}$ cells) was added along with $1.25\textrm{(OH)}_2\textrm{D}_3$. Ionophore($0.05\mu\textrm{g}$/$10^{6}$ cells) alone increased specifically the value of the slow turnover rate which was not affected by addition of diltiazem. The hypothesis concerning the involvement of calcium in bone resorption seems in fact to be verified in case of $1.25\textrm{(OH)}_2\textrm{D}_3$ but more unsettled for Ca inophore A23187.

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Potentiometric performances of polymer membrane electrode based on cyclosporin (싸이크로스포린을 이용한 고분자막 전위차 전극)

  • Rhee Paeng, Insook
    • Analytical Science and Technology
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    • v.18 no.6
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    • pp.491-494
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    • 2005
  • The main component governing selectivity in ion-selective electrodes and optodes is the ionophore. For this reason, a member of natural products that possess selective ion-binding properties have long been sought after. By applying this principle, the performance of cyclosporin used as neutral carriers for calcium selective polymeric membrane electrode was investigated. The calcium ion-selective electrode based on cyclosporin gave a good Nernstian response of 26.6 mV per decade for calcium ion in the activity range $1{\times}10^{-6}M$ to $1{\times}10^{-2}M$. The optimized calcium ion-selective electrode displayed very comparable selectivity for $Ca^{2+}$ ion against alkali and alkaline earth metal ions, $Na^{2+}$, and $Mg^{2+}$ in particular.

Ionophore Activity of Frangufoline

  • Park, Man-Ki;Park, Jeong-Hill;Cho, Jung-Hwan;Park, Jun-Kyu;Han, Yong-Nam;Han, Byung-Hoon
    • Archives of Pharmacal Research
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    • v.14 no.2
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    • pp.103-104
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    • 1991
  • The ionophore activity of frangufoline (1), a sedative cyclopeptide alkaloid isolated from Zizyphus jujuba, was investigated by UV and CD spectroscopic methods. Frangufoline (1) showed ion binding activity to calcium and magnesium ions.

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Efficient Establishment of Presumptive Embryonic Stem Cells from Bovine Blastocysts by Exposure to Calcium Ionophore (Calcium Ionophore를 이용한 소 배반포로 부터의 배아주 유사세포의 효과적인 분리)

  • 김선욱;류재웅;이철상;한용만;박정선;유대열;이경광
    • Development and Reproduction
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    • v.3 no.1
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    • pp.45-51
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    • 1999
  • It is crucial to remove trophectoderm (TE) cells of blastocysts for an efficient isolation of pluripotent embryonic stem (ES)-like cells from bovine blastocysts. We evaluated the effectiveness of chemosurgery using calcium ionophore A23l87 (CIPA) by investigating the viability and pluripotency of ES-like cell lines isolated from in vitro-produced bovine blastocysts after CIPA treatment. The blastocysts treated with 50 $\mu$M CIPA for 25 min colonized most efficiently (51% of blastocysts) and developed to ES-like cell lines through 10 passages (4.8% of blastocysts) among CIPA-treated groups with different concentration and duration. In comparison with CIPA-untreated blastocysts, the colonization rate and overall viability of the CIPA-treated blastocysts were five times higher, suggesting that CIPA treatment condition defined in this study was highly efficient for establishing ES-like cell lines without apparent toxicity of CIPA. We evaluated in vitro pluripotency of the established three ES-like cell lines by examining alkaline phosphatase (AP) activity, capability of embryoid body formation, and chromosomal euploidity of the cells. Our cells showed a heterogeneous AP activity similarly to other reports. The cells were able to form simple embryoid bodies during suspension culture and majority of them showed a normal chromosome number of 60, the euploid chromosomal complement of bovine Therefore, our data suggest that CIPA treatment can be safely used for an efficient isolation of ES-like cell lines from bovine blastocysts.

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Structural Organization of Calmodulin Gene and Expression in Transgenic

  • 최영주
    • Journal of Life Science
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    • v.4 no.2
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    • pp.50-59
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    • 1994
  • 신호전달과정의 연구는 calcium이 messenger로서 작용한다고 밝혀진 후로 식물에서 $Ca^{++}$ -messenger system에 대한 생화학적 및 분자생물학적 분야에서의 연구는 급속하게 발전하게 되었다. 식물세포에서 calcium 이온들의 많은 작용은 EF hand family로서 알려진 calcium binding protein에 의해서 조절된다. Calmodulin (CaM)은 highy conserve 되어 있으며, 4개의 calcium binding domain을 가진 ubiquitous한 단백질이다. 본 연구는 calmodulin 유전자의 발현에 미치는 calcium, EGTA, calcium ionophore 및 calmodulin antagonist의 영향과 또한 외부신호(light, wounding), chemical 및 auxin 등의 영향을 reporter화 유전자의 분석에 의해서 CaM유전자의 발현기작을 규명하고자 하였고, 또한 calmodulin 유전자의 organ-specific 발현 및 calmodulin의 새로운 생리적인 기능도 연구하고자 하였다.

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Effects of Calcium Ionophore on Vitellogenin Production in the Culture of Hepatocytes in the Rinbow Trout, Oncorhynchus mykess (무지개송어(Oncorhynchus mykiss) 배양 간세포에서 Vitellogenin 합성에 미치는 Calcium ionophore의 영향)

  • 여인규
    • Journal of Aquaculture
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    • v.11 no.2
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    • pp.241-248
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    • 1998
  • Effects of A23187 on estradiol$-17^{\beta}$-induced vitellogenin (VTG) induction were electrophore-tically examined in primary hepatocyte cultures in rainbow trout. hepatocytes were predultured for 2 days and then estradiol-$17^{\beta}$(E2, $2{\times}10^{-6}$M) and calcium ionophore (A23187, $10^{-7)$~$10^{-5}$ M) were added to the incubation medium. The hepatocytes were cultured for 7 more days. In addition, effects of A23187 on $E_2$-primed VTG production were investigated for 7 days. The addition of A23187 ($10^{-7)$~$10^{-5}$M) to the incubation medium specifically reduced VTG production by hepatocytes in a concentration-dependent way. The addition of A23187 significantly reduced the rate of $E_2$-primed VTG production to 18% of the control (E2 only) on Day 7. However, $E_2$-primed VTG production was reduced to 47% of the control by withdrawal of $E_2$ from the incubation medium. Therefore, these results suggest that intracellualr sequestered calcium could regulate VTG synthesis at the translational and/or post-translational stage.

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Artificial oocyte activation in intracytoplasmic sperm injection cycles using testicular sperm in human in vitro fertilization

  • Kang, Hee Jung;Lee, Sun-Hee;Park, Yong-Seog;Lim, Chun Kyu;Ko, Duck Sung;Yang, Kwang Moon;Park, Dong-Wook
    • Clinical and Experimental Reproductive Medicine
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    • v.42 no.2
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    • pp.45-50
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    • 2015
  • Objective: Artificial oocyte activation (AOA) is an effective method to avoid total fertilization failure in human in vitro fertilization-embryo transfer (IVF-ET) cycles. AOA performed using a calcium ionophore can induce calcium oscillation in oocytes and initiate the fertilization process. We evaluated the usefulness of AOA with a calcium ionophore in cases of total fertilization failure in previous cycles and in cases of severe male factor infertility patients with non-motile spermatozoa after pentoxifylline (PF) treatment. Methods: The present study describes 29 intracytoplasmic sperm injection (ICSI)-AOA cycles involving male factor infertility at Cheil General Hospital from January 2006 to June 2013. Patients were divided into two groups (control, n=480; AOA, n=29) depending on whether or not AOA using a calcium ionophore (A23187) was performed after testicular sperm extraction-ICSI (TESE-ICSI). The AOA group was further split into subgroups according to sperm motility after PF treatment: i.e., motile sperm-injected (n=12) and non-motile sperm-injected (n=17) groups (total n=29 cycles). Results: The good embryo rate (52.3% vs. 66.9%), pregnancy rate (20.7% vs. 52.1%), and delivery rate (10.3% vs. 40.8%) were lower in the PF/AOA group than in the control group. When evaluating the effects of restoration of sperm motility after PF treatment on clinical outcomes there was no difference in fertilization rate (66.6% vs. 64.7% in non-motile and motile sperm, respectively), pregnancy rate (17.6% vs. 33.3%), or delivery rate (5.9% vs. 16.7%) between the two groups. Conclusion: We suggest that oocyte activation is a useful method to ensure fertilization in TESE-ICSI cycles regardless of restoration of sperm motility after PF treatment. AOA may be useful in selected patients who have a low fertilization rate or total fertilization failure.