• Title/Summary/Keyword: Ion channels

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Englerin A-sensing charged residues for transient receptor potential canonical 5 channel activation

  • Jeong, SeungJoo;Ko, Juyeon;Kim, Minji;Park, Ki Chul;Park, Eunice Yon June;Kim, Jinsung;Baik, Youngjoo;Wie, Jinhong;Cho, Art E.;Jeon, Ju-hong;So, Insuk
    • The Korean Journal of Physiology and Pharmacology
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    • v.23 no.3
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    • pp.191-201
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    • 2019
  • The transient receptor potential canonical (TRPC) 5 channel, known as a nonselective cation channel, has a crucial role in calcium influx. TRPC5 has been reported to be activated by muscarinic receptor activation and extracellular pH change and inhibited by the protein kinase C pathway. Recent studies have also suggested that TRPC5 is extracellularly activated by englerin A (EA), but the mechanism remains unclear. The purpose of this study is to identify the EA-interaction sites in TRPC5 and thereby clarify the mechanism of TRPC5 activation. TRPC5 channels are over-expressed in human embryonic kidney (HEK293) cells. TRPC5 mutants were generated by site-directed mutagenesis. The whole-cell patch-clamp configuration was used to record TRPC5 currents. Western analysis was also performed to observe the expression of TRPC5 mutants. To identify the EA-interaction site in TRPC5, we first generated pore mutants. When screening the mutants with EA, we observed the EA-induced current increases of TRPC5 abolished in K554N, H594N, and E598Q mutants. The current increases of other mutants were reduced in different levels. We also examined the functional intactness of the mutants that had no effect by EA with TRPC5 agonists, such as carbachol or $GTP{\gamma}S$. Our results suggest that the three residues, Lys-554, His-594, and Glu-598, in TRPC5 might be responsible for direct interaction with EA, inducing the channel activation. We also suggest that although other pore residues are not critical, they could partly contribute to the EA-induced channel activation.

Regulatory mechanisms of the store-operated Ca2+ entry through Orai1 and STIM1 by an adaptor protein in non-excitable cells

  • Kang, Jung Yun;Yang, Yu-Mi
    • International Journal of Oral Biology
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    • v.47 no.3
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    • pp.33-40
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    • 2022
  • Store-operated Ca2+ entry (SOCE) represents one of the major Ca2+ entry routes in non-excitable cells. It is involved in a variety of fundamental biological processes and the maintenance of Ca2+ homeostasis. The Ca2+ release-activated Ca2+ (CRAC) channel consists of stromal interaction molecule and Orai; however, the role and action of Homer proteins as an adaptor protein to SOCE-mediated Ca2+ signaling through the activation of CRAC channels in non-excitable cells still remain unknown. In the present study, we investigated the role of Homer2 in the process of Ca2+ signaling induced by the interaction between CRACs and Homer2 proteins in non-excitable cells. The response to Ca2+ entry by thapsigargin-mediated Ca2+ store depletion remarkably decreased in pancreatic acinar cells of Homer2-/- mice, as compared to wild-type cells. It also showed critical differences in regulated patterns by the specific blockers of SOCE in pancreatic acinar cells of Homer2-/- mice. The response to Ca2+ entry by the depletion in Ca2+ store markedly increased in the cellular overexpression of Orai1 and STIM1 as compared to the overexpression of Homer2 in cells; however, this response was remarkably inhibited by the overexpression of Orai1, STIM1, and Homer2. These results suggest that Homer2 has a critical role in the regulatory action of SOCE activity and the interactions between CRAC channels.

Effect of Humidity and Flooding on the Performance of Proton Exchange Membrane Fuel Cell (고분자전해질 연료전지의 성능에 미치는 습도와 플러딩의 영향)

  • Hwang, Byungchan;Chung, Hoi-Bum;Song, Myung-Hyun;Oh, Sung-June;Na, Il-Chai;Park, Kwonpil
    • Korean Chemical Engineering Research
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    • v.55 no.3
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    • pp.302-306
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    • 2017
  • Humidity affect performance and durability of proton exchange membrane fuel cell (PEMFC). High humidity of gases generally enhance the performance, but high humidity have the danger of flooding. I-V performance, linear sweep voltammetry, cyclo voltammetry, and impedance of micro-channel cell measured with change of relative humidity (RH). Flooding phenomena started at RH 70%. Ion conductivity of membrane reached maximum value at RH 80%. Maximum current density of $1,700mA/cm^2$ (at 0.6 V) was obtained at RH 80%. Therefore the effect of ion conductivity increasement was higher than that of mass transfer decrease by flooding at RH 80%.

Control of $Ca^{2+}$- Influx by $Ca^{2+}$/Calmodulin Dependent Protein Kinase II in the Activation of Mouse Eggs

  • Yoon, Sook-Young;Kang, Da-Won;Bae, In-Ha
    • Development and Reproduction
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    • v.15 no.1
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    • pp.31-39
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    • 2011
  • Change in intracellular $Ca^{2+}$-concentration ($[Ca^{2+}]_i$) is an essential event for egg activation and further development. $Ca^{2+}$ ion is originated from intracellular $Ca^{2+}$-store via inositol 1,4,5-triphosphate receptor and/or $Ca^{2+}$ influx via $Ca^{2+}$ channel. This study was performed to investigate whether changes in $Ca^{2+}$/calmodulin dependent protein kinase II (CaM KII) activity affect $Ca^{2+}$ influx during artificial egg activation with ethanol using $Ca^{2+}$ monitoring system and whole-cell patch clamp technique. Under $Ca^{2+}$ ion-omitted condition, $Ca^{2+}$-oscillation was stopped within 30 min post microinjection of porcine sperm factor, and ethanol-induced $Ca^{2+}$ increase was reduced. To investigate the role of CaM KII known as an integrator of $Ca^{2+}$- oscillation during mammalian egg fertilization, CaM KII activity was tested with a specific inhibitor KN-93. In the eggs treated with KN-93, ethanol failed to induce egg activation. In addition, KN-93 inhibited inward $Ca^{2+}$ current ($I_{Ca}$) in a time-dependent manner in whole-cell configuration. Immunostaining data showed that the voltage-dependent $Ca^{2+}$ channels were distributed along the plasma membrane of mouse egg and 2-cell embryo. From these results, we suggest that $Ca^{2+}$ influx during fertilization might be controlled by CaM KII activity.

Comparison of electrophysiological effects of calcium channel blockers on cardiac repolarization

  • Lee, Hyang-Ae;Hyun, Sung-Ae;Park, Sung-Gurl;Kim, Ki-Suk;Kim, Sung Joon
    • The Korean Journal of Physiology and Pharmacology
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    • v.20 no.1
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    • pp.119-127
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    • 2016
  • Dihydropyridine (DHP) calcium channel blockers (CCBs) have been widely used to treat of several cardiovascular diseases. An excessive shortening of action potential duration (APD) due to the reduction of $Ca^{2+}$ channel current ($I_{Ca}$) might increase the risk of arrhythmia. In this study we investigated the electrophysiological effects of nicardipine (NIC), isradipine (ISR), and amlodipine (AML) on the cardiac APD in rabbit Purkinje fibers, voltage-gated $K^+$ channel currents ($I_{Kr}$, $I_{Ks}$) and voltage-gated $Na^+$ channel current ($I_{Na}$). The concentration-dependent inhibition of $Ca^{2+}$ channel currents ($I_{Ca}$) was examined in rat cardiomyocytes; these CCBs have similar potency on $I_{Ca}$ channel blocking with $IC_{50}$ (the half-maximum inhibiting concentration) values of 0.142, 0.229, and 0.227 nM on NIC, ISR, and AML, respectively. However, ISR shortened both $APD_{50}$ and $APD_{90}$ already at $1{\mu}M$ whereas NIC and AML shortened $APD_{50}$ but not $APD_{90}$ up to $30{\mu}M$. According to ion channel studies, NIC and AML concentration-dependently inhibited $I_{Kr}$ and $I_{Ks}$ while ISR had only partial inhibitory effects (<50% at $30{\mu}M$). Inhibition of $I_{Na}$ was similarly observed in the three CCBs. Since the $I_{Kr}$ and $I_{Ks}$ mainly contribute to cardiac repolarization, their inhibition by NIC and AML could compensate for the AP shortening effects due to the block of $I_{Ca}$.

Formation Behavior of Microporous Ettringite Body by Hydration of Tricalciumaluminate Clinker and Gypsum (알루민산삼칼슘 클링커와 석고의 수화에 의한 에트린자이트 미세다공체 생성거동)

  • Na, Hyeon-Yeop;Song, Tae-Woong
    • Journal of the Korean Ceramic Society
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    • v.45 no.11
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    • pp.734-738
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    • 2008
  • Ettringite$(3CaO{\cdot}Al_2O_3{\cdot}3CaSO_4{\cdot}32H_2O)$, one of the hydrated phase of Portland cement is usually formed in the early stage of hydration by the reaction of tricalciumaluminate$(C_3A)$ and gypsum. The rapid and strong crystal growth of separated rod-shaped ettringite have been utilized for the preparation of special cements of rapid setting, high strength and non-shrinking properties. The ettringite also has been noticed as a promising materials for the immobilization of various waste ions because of its unique crystal structure which has abundant channels and exchangeable ionic compounds. In this study, the formation and growth behavior of the ettringite was investigated in the system $C_3A-CaSO_4-H_2O$ using $C_3A$ clinker and gypsum to obtain a microporous body for waste ion immobilization. Ettringite was revealed to form by the dissolution-precipitation mechanism and the bulk body was by the entangled growth of rod-shaped ettringite crystals. The hardened body was composed of nearly pure rod-shaped ettringite interlocked each other with adequate mechanical strength. The homogeneity of structure, pore size, specific surface area and porosity of the hardened body were influenced by reaction temperature, water/powder ratio and the curing time. The hardened body prepared with water/powder ratio of 1 at $24^{\circ}C$ for one day showed excellent morphological properties for the purposed materials.

Photodissociation of C3H5Br and C4H7Br at 234 nm

  • Kim, Hyun-Kook;Paul, Dababrata;Hong, Ki-Ryong;Cho, Ha-Na;Lee, Kyoung-Seok;Kim, Tae-Kyu
    • Bulletin of the Korean Chemical Society
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    • v.33 no.1
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    • pp.143-148
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    • 2012
  • The photodissociation dynamics of cyclopropyl bromide ($C_3H_5Br$) and cyclobutyl bromide ($C_4H_7Br$) at 234 nm was investigated. A two-dimensional photofragment ion-imaging technique coupled with a [2+1] resonanceenhanced multiphoton ionization scheme was utilized to obtain speed and angular distributions of the nascent $Br(^2P_{3/2})$ and $Br^*(^2P_{1/2})$ atoms. The recoil anisotropies for the Br and $Br^*$ channels were measured to be ${\beta}_{Br}=0.92{\pm}0.03$ and ${\beta}_{Br^*}=1.52{\pm}0.04$ for $C_3H_5Br$ and ${\beta}_{Br}=1.10{\pm}0.03$ and ${\beta}_{Br^*}=1.49{\pm}0.05$ for $C_4H_7Br$. The relative quantum yield for Br was found to be ${\Phi}_{Br}=0.13{\pm}0.03$ and for $C_3H_5Br$ and $C_4H_7Br$, respectively. The soft radical limit of the impulsive model adequately modeled the related energy partitioning. The nonadiabatic transition probability from the 3A' and 4A' potential energy surfaces was estimated and discussed.

Effects study of Aconiti Lateralis Radix Preparata extract on the regulation of heat and cold in PTU-induced hypothyroidism rats (포부자추출물의 갑상선기능저하증 흰쥐모델에서의 한열조절작용에 의한 개선효능 연구)

  • Hwang, Min Sub;Hwang, Ji Hye;Kang, Seok Yong;Kang, An Na;Roh, Hyo Sun;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.31 no.6
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    • pp.63-71
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    • 2016
  • Objective : To suggest a scientific evidence of Aconitum carmichaeli Debx. (Aconiti Lateralis Radix Preparata: ALRP) as one of cooling and heating medicines on the regulation of body temperature, we investigated the effects of ALRP water extract on hypothyroidism. Methods : Hypothyroidism was induced by intradermal injection with PTU for 4 weeks in SD rats. ALRP extract or L-thyroxine as a control drug was orally administrated for 2 weeks with PTU injection in rats. The physiological and serological parameters were measured in rats. The histological change of thyroid tissues was observed by H&E staining, and also the expression of thermo-regulating proteins was determined by Western blot in dorsal root ganglia and brain tissues of rats. Results : The administration of ALRP extract in PTU-induced hypothyroidism rats was significantly increased body temperature, but did not changes on body weight, food and water intake. ALRP extract did not effect on the levels of TSH and T4 in the hypothyroidism rats. ALRP extract significantly decreased the levels of GPT, glucose, HDL-cholesterol, LDL-cholesterol, and total cholesterol in the hypothyroidism rats. In histological observation, the enlarged epithelium and atrophic follicles with higher concentration of follicular cells on hypothyroidism were improved by ALRP extract. In addition, ALRP extract increased the expression of TRPV1 and TRPM8 ion channel proteins in hypothyroidism rats. Conclusion : These results indicate that ALRP extract can improve PTU-induced hypothyroidism through regulation of body temperature and lipid accumulation. The action mechanism of ALRP extract is related with body temperature control by thermoregulation with TRP ion channels.

Polymer Electrolyte Membranes of Poly(Styrene-Butadiene-Styrene) Star Triblock Copolymer for Fuel Cell (연료전지용 Poly(Styrene-Butadiene-Styrene) Star Triblock Copolymer의 고분자 전해질 분리막)

  • Garcia, Edwin D.;Jung, Bumsuk
    • Membrane Journal
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    • v.29 no.5
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    • pp.252-262
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    • 2019
  • A sulfonated star branched poly(styrene-b-butadiene-b-styrene) triblock copolymer (SSBS) was synthesized with varying degrees of sulfonation. The effective sulfonation on the butadiene block was confirmed by FT-IR spectroscopy. Ion exchange capacity by titration was used to determine the degree of sulfonation. The synthesized polymer observed enhanced water uptake and proton conductivity. At room temperature, the SSBS with 25 mol% degree of sulfonation showed an outstanding proton conductivity of 0.114 S/cm, similar to that of commercial membrane, Nafion. The effect of temperature at constant relative humidity on conductivity resulted to a remarkable increase in proton conductivity. Methanol permeability studies showed a value lower than Nafion for all the sulfonated membranes. Structural nature observed using AFM showed that the membranes observed microphase separated nanostructures and the connectivity of the interionic channels.

Influence of Fimasartan (a Novel $AT_1$ Receptor Blocker) on Catecholamine Release in the Adrenal Medulla of Spontaneously Hypertensive Rats

  • Lim, Hyo-Jeong;Lee, Seog-Ki;Lim, Dong-Yoon
    • The Korean Journal of Physiology and Pharmacology
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    • v.17 no.1
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    • pp.99-109
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    • 2013
  • The aim of this study was to determine whether fimasartan, a newly developed $AT_1$ receptor blocker, can affect the CA release in the isolated perfused model of the adrenal medulla of spontaneously hypertensive rats (SHRs). Fimasartan (5~50 ${\mu}M$) perfused into an adrenal vein for 90 min produced dose- and time-dependently inhibited the CA secretory responses evoked by ACh (5.32 mM), high $K^+$ (56 mM, a direct membrane depolarizer), DMPP (100 ${\mu}M$) and McN-A-343 (100 ${\mu}M$). Fimasartan failed to affect basal CA output. Furthermore, in adrenal glands loaded with fimasartan (15 ${\mu}M$), the CA secretory responses evoked by Bay-K-8644 (10 ${\mu}M$, an activator of L-type $Ca^{2+}$ channels), cyclopiazonic acid (10 ${\mu}M$, an inhibitor of cytoplasmic $Ca^{2+}$-ATPase), and veratridine (100 ${\mu}M$, an activator of $Na^+$ channels) as well as by angiotensin II (Ang II, 100 nM), were markedly inhibited. In simultaneous presence of fimasartan (15 ${\mu}M$) and L-NAME (30 ${\mu}M$, an inhibitor of NO synthase), the CA secretory responses evoked by ACh, high $K^+$, DMPP, Ang II, Bay-K-8644, and veratridine was not affected in comparison of data obtained from treatment with fimasartan (15 ${\mu}M$) alone. Also there was no difference in NO release between before and after treatment with fimasartan (15 ${\mu}M$). Collectively, these experimental results suggest that fimasartan inhibits the CA secretion evoked by Ang II, and cholinergic stimulation (both nicotininc and muscarinic receptors) as well as by membrane depolarization from the rat adrenal medulla. It seems that this inhibitory effect of fimasartan may be mediated by blocking the influx of both $Na^+$ and $Ca^{2+}$ through their ion channels into the rat adrenomedullary chromaffin cells as well as by inhibiting the $Ca^{2+}$ release from the cytoplasmic calcium store, which is relevant to $AT_1$ receptor blockade without NO release.