• Title/Summary/Keyword: $Ca^{2+}$ transient

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Deficiency of Anoctamin 5/TMEM16E causes nuclear positioning defect and impairs Ca2+ signaling of differentiated C2C12 myotubes

  • Phuong, Tam Thi Thanh;An, Jieun;Park, Sun Hwa;Kim, Ami;Choi, Hyun Bin;Kang, Tong Mook
    • The Korean Journal of Physiology and Pharmacology
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    • v.23 no.6
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    • pp.539-547
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    • 2019
  • Anoctamin 5 (ANO5)/TMEM16E belongs to a member of the ANO/TMEM16 family member of anion channels. However, it is a matter of debate whether ANO5 functions as a genuine plasma membrane chloride channel. It has been recognized that mutations in the ANO5 gene cause many skeletal muscle diseases such as limb girdle muscular dystrophy type 2L (LGMD2L) and Miyoshi muscular dystrophy type 3 (MMD3) in human. However, the molecular mechanisms of the skeletal myopathies caused by ANO5 defects are poorly understood. To understand the role of ANO5 in skeletal muscle development and function, we silenced the ANO5 gene in C2C12 myoblasts and evaluated whether it impairs myogenesis and myotube function. ANO5 knockdown (ANO5-KD) by shRNA resulted in clustered or aggregated nuclei at the body of myotubes without affecting differentiation or myotube formation. Nuclear positioning defect of ANO5-KD myotubes was accompanied with reduced expression of Kif5b protein, a kinesin-related motor protein that controls nuclear transport during myogenesis. ANO5-KD impaired depolarization-induced $[Ca2^{+}]_i$ transient and reduced sarcoplasmic reticulum (SR) $Ca^{2+}$ storage. ANO5-KD resulted in reduced protein expression of the dihydropyridine receptor (DHPR) and SR $Ca^{2+}-ATPase$ subtype 1. In addition, ANO5-KD compromised co-localization between DHPR and ryanodine receptor subtype 1. It is concluded that ANO5-KD causes nuclear positioning defect by reduction of Kif5b expression, and compromises $Ca^{2+}$ signaling by downregulating the expression of DHPR and SERCA proteins.

Application of Semi-continuous Ambient Aerosol Collection System for Elemental Analysis (대기입자의 원소성분 배출특성연구를 위한 반-연속식 입자채취시스템 적용)

  • Park, Seung-Shik;Ko, Jae-Min;Lee, Dong-Soo
    • Journal of Korean Society for Atmospheric Environment
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    • v.28 no.1
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    • pp.39-51
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    • 2012
  • Aerosol slurry samples were collected in 60-min interval using Korean Semi-continuous Elements in Aerosol Sampler (KSEAS) between May 19 and June 6, 2010 at an urban site of Gwangju. The $PM_{2.5}$ samples were collected with a flow rate of 16.7 L/min and particles are grown by condensation of water vapor in a condenser maintained at ${\sim}5^{\circ}C$ after saturation by direct injection of steam. The resulting droplets are collected in a liquid slurry with a airdroplet separator. Concentrations of 16 elements (Al, Fe, Mn, Ca, K, Cu, Zn, Pb, Cd, Cr, Ti, V, Ni, Co, As, Se) in the collected slurry samples were determined off-line by ICP-MS. KSEAS sample analysis encompassed the sampling periods for which 24-hr average elemental species concentrations were calculated for comparison with those derived from 24-hr integrated filter samples. Relationship between elemental species measured by two methods indicated high correlation coefficients (r), mostly greater than r of 0.80. However, we note that concentrations of Al, K, Ca, Mn, and Fe, which are often associated with crustal elemental particles, in the KSEAS samples, were substantially lower (1.4~11 times) than those found in the typical filter-based samples. This discrepancy is probably due to difficulties in transferring insoluble dust particles to the collection vials in the KSEAS. Temporal profiles of elemental concentrations indicate that some transient events in their concentrations are observed over the sampling periods. For the elemental species studied, atmospheric concentrations during the transient events increased by factors of 4 in Mn~80 in Zn, compared to their background levels. Principle component analyses were applied to the hourly KSEAS data sets to identify sources affecting the concentrations of the metal constituents observed. In this study, we conclude that hourly measurements for particle-bound elemental constituents were extremely useful for revealing the short-term variability in their concentrations and developing insights into their sources.

Mutation of a putative S-nitrosylation site of TRPV4 protein facilitates the channel activates

  • Lee, Eun-Jeoung;Shin, Sung-Hwa;Hyun, Sung-Hee;Chun, Jae-Sun;Kang, Sang-Sun
    • Animal cells and systems
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    • v.15 no.2
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    • pp.95-106
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    • 2011
  • The transient receptor potential vanilloid 4 (TRPV4) cation channel, a member of the TRP vanilloid subfamily, is expressed in a broad range of tissues. Nitric oxide (NO) as a gaseous signal mediator shows a variety of important biological effects. In many instances, NO has been shown to exhibit its activities via a protein S-nitrosylation mechanism in order to regulate its protein functions. With functional assays via site-directed mutagenesis, we demonstrate herein that NO induces the S-nitrosylation of TRPV4 $Ca^{2+}$ channel on the $Cys^{853}$ residue, and the S-nitrosylation of $Cys^{853}$ reduced its channel sensitivity to 4-${\alpha}$ phorbol 12,13-didecanoate and the interaction between TRPV4 and calmodulin. A patch clamp experiment and $Ca^{2+}$ image analysis show that the S-nitrosylation of $Cys^{853}$ modulates the TRPV4 channel as an inhibitor. Thus, our data suggest a novel regulatory mechanism of TRPV4 via NO-mediated S-nitrosylation on its $Cys^{853}$ residue.

Novel Glycolipoproteins from Ginseng

  • Pyo, Mi-Kyung;Choi, Sun-Hye;Hwang, Sung-Hee;Shin, Tae-Joon;Lee, Byung-Hwan;Lee, Sang-Mok;Lim, Yoong-Ho;Kim, Dong-Hyun;Nah, Seung-Yeol
    • Journal of Ginseng Research
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    • v.35 no.1
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    • pp.92-103
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    • 2011
  • Ginseng has been used as a general tonic agent to invigorate human body. In the present study, we isolated novel glycolipoproteins from ginseng that activate $Ca^{2+}$-activated $Cl^-$ channel (CaCC) in Xenopus oocytes and transiently increase intracellular free $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) in mouse Ehrlich ascites tumor cells. We named the active ingredients as gintonin. Gintonin exists in at least six different forms. The native molecular weight of gintonin is about 67 kDa but its apparent molecular weight is about 13 kDa, indicating that gintonin might be a pentamer. Gintonin is rich in hydrophobic amino acids. Its main carbohydrates are glucose and glucosamine. Its lipid components are linoleic, palmitic, oleic, and stearic acids. Gintonin actions were blocked by U73122, a phospholipase C inhibitor, 2-aminoethxydiphenyl borate, an inositol 1,4,5-trisphosphate receptor antagonist, or bis (o-aminophenoxy) ethane-N,N,N0,N0-tetracetic acid acetoxymethyl ester, a membrane permeable $Ca^{2+}$ chelator. In the present study, we for the first time isolated novel gintonin and showed the signaling pathways on gintonin-mediated CaCC activations and transient increase of $[Ca^{2+}]_i$. Since $[Ca^{2+}]_i$ as a second messenger plays a pivotal role in the regulation of diverse $Ca^{2+}$-dependent intracellular signal pathways, gintonin-mediated regulations of $[Ca^{2+}]_i$ might contribute to biological actions of ginseng.

Effects of $Ca^{++}$, Verapamil and $La^{+++}$ on the Spontaneous Contraction and K-contracture in the Isolated Rat Uterine Smooth Muscle (칼슘, 베라파밀, 란타눔이 흰쥐 자궁근의 자발적 수축과 칼륨 경축에 미치는 효과)

  • Hwang, Sang-Ik
    • The Korean Journal of Physiology
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    • v.18 no.1
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    • pp.37-50
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    • 1984
  • The effects of $Ca^{++}$ and its antagonists (verapamil and $La^{+++}$) upon the spontaneous contraction and the contracture induced by 60 mM K-Tyrode solution were studied in the isolated uterine muscle. Longitudinal muscle strips were prepared from the rat uteri at estrous stage. All experiments were performed in tris-buffered Tyrode solution which was aerated with 100% $O_2$ and kept at 35^{\circ}$. The results obtained were as follows: 1) In the uterine strips contracting spontaneously, both the amplitude of peak tension and the area of contraction curve increased dose-dependently in the range of $0.5${\sim}8$ mM $Ca^{++}$. The frequency of contraction increased as the concentration of $Ca^{++}$ increased up to 2 mM, but above this concentration the frequency decreased. In $Ca^{++}-free$ media, however, contraction did not develop. In the contracture induced by 60 mM K-Tyrode solution, the developed tension increased dose-dependently as the concentration of external $Ca^{++}$ increased to 8 mM. In the absence of external $Ca^{++}$ K-contracture appeared, but it was not sustained. 2) The spontaneous contraction of rat uterus was suppressed by verapamil in proportion to an increase of its concentration and totally abolished at the concentration of $3{\times}10^{-4}\;g/l$, but the spontaneous contraction re-appeared by addition of $Ca^{++}$. The amplitude of peak tension recovered completely but the recovery of frequency was incomplete. K-contracture decreased in a dose-dependent manner after the treatment with verapamil and totally disappeared at its concentration of $3{\times}10^{-4}\;g/l$. Even in this case contracture developed again by extra $Ca^{++}$. 3) The spontaneous contractile activity was inhibited by $La^{+++}$. At the concentration of $10^{-4}$M $La^{+++}$, fibrillation appeared. In the strip inhibited by $10^{-5}M\;La^{+++}$, contractility recovered completely by extra $Ca^{++}$ while in the $10^{-4}M\;La^{+++}$ treated preparation, the rhythmic spontaneous contraction did not develop even at the concentration of 16 mM $Ca^{++}$. After the initial transient depression of contracture tension by $10^{-3}M$ of $La^{+++}$, the strip stowed considerably large size of contracture, hardly influenced by external $Ca^{++}$ or verapamil. The results obtained in this experiment suggest that in the rat uterine muscle there would be some competitive actions between $Ca^{++}$ and its antagonists. It is speculated that $Ca^{++}$ plays an important role in the conduction of excitation, and $La^{+++}$ influences upon cellular $Ca^{++}$ mobilization and re-uptake process as well as transmembrane $Ca^{++}$ transport in a K-depolarized state.

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Cancelling Magnetic Features on the Sun

  • Park, So-Young;Chae, Jong-Chul
    • The Bulletin of The Korean Astronomical Society
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    • v.36 no.1
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    • pp.36.2-36.2
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    • 2011
  • A cancelling magnetic feature (CMF) is believed to be a result of magnetic reconnection in the low atmosphere of the Sun. In this work, we investigate the physical properties of CMFs, focusing on the rates of flux cancellation in CMFs and the dynamics of chromospheric phenomena coupled with CMFs. First, we have determined the specific rates of flux cancellation using the magnetograms taken by the Solar Optical Telescope (SOT) aboard the Hinode satellite. The specific rates determined with the SOT turned out to be systematically higher than those based on the data taken by the Michelson Doppler Imager (MDI) aborad the SOHO. Second, we analyzed transient Ca II brightenings associated with small-scale CMFs using the SOT/Hinode. We found that in most Ca II brightenings related to CMFs, and the Ca II intensity peaks after magnetic flux cancellation proceeds. Moreover, brightenings tend to appear as pairs of bright points of similar size and similar brightness overlying magnetic bipoles. To further study the brightening and dynamics of chromospherie features associated with CMFs, we have analyzed Fast Imaging Solar Spectrograph (FISS) data. From this data the Doppler motion of chromospheric features above a CMF changed from redshift to blueshift. The duration of such dynamics is very short being less than 2 minutes. These results are unexpected one and can not be explained by any pre-existing pictures of CMFs.

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Insect GPCRs and TRP Channels: Putative Targets for Insect Repellents

  • Kim, Sang Hoon
    • Interdisciplinary Bio Central
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    • v.5 no.3
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    • pp.6.1-6.7
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    • 2013
  • Many insects such as mosquitoes cause life-threatening diseases such as malaria, yellow fever and West Nile virus. Malaria alone infects 500 million people annually and causes 1-3 million death per year. Volatile insect repellents, which are detected through the sense of smell, have long been used to protect humans against insect pests. Antifeed-ants are non-volatile aversive compounds that are detected through the sense of taste and prevent insects from feeding on plants. The molecular targets and signaling path-ways required for sensing insect repellents and antifeedants are poorly understood. Transient Receptor Potential (TRP) Ca2+-permeable cation channels exist in organisms ranging from C. elegans to D. melanogaster and Homo sapiens. Drosophila has 13 family members, which mainly function in sensory physiology such as vision, thermotaxis and chemotaxis. G protein-coupled receptors (GPCRs) initiate olfactory signaling cascades in mammals and in nematodes C.elegans. However, the mechanisms of G protein signaling cascades in insect chemosensation are controversial. In this review, I will discuss the putative roles of G protein-coupled receptors (GPCRs) and Transient Receptor Potential (TRP) channels as targets for insect repellents.

The role of ginseng total saponin in transient receptor potential melastatin type 7 channels

  • Kim, Byung Joo
    • Animal cells and systems
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    • v.16 no.5
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    • pp.376-384
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    • 2012
  • Although ginsenosides have a variety of physiologic or pharmacologic functions in various regions, there are only a few reports on the effects of transient receptor potential melastatin 7 (TRPM7) channels. Here, we showed evidence suggesting that TRPM7 channels play an important role in ginseng total saponin (GTS)-mediated cellular injury. The combination techniques of electrophysiology, pharmacological analysis, small interfering RNA (siRNA) method and cell death assays were used. GTS depolarized the resting membrane potentials and decreased the amplitude of pacemaker potentials in cultured interstitial cells of Cajal (ICCs) in gastrointestinal (GI) tract. The TRPM7-like currents in single ICCs and the overexpressing TRPM7 in HEK293 cells were inhibited by GTS. However, GTS had no effect on $Ca^{2+}$-activated $Cl^-$ conductance. GTS inhibited the survival of human gastric (AGS) and brea (MCF-7) adenocarcinoma cells. Also, GTS inhibited the TRPM7-like currents in AGS and MCF-7 cells. The GTS-mediated cytotoxicity was inhibited by TRPM7-specific siRNA. In addition, we showed that overexpression of TRPM7 channels in HEK293 cells was inhibited by GTS. Thus, TRPM7 channels are involved in GTS-mediated cell death in AGS and MCF-7 cells, and these channels may represent a novel target for physiological disorders where GTS plays an important role.

Time-resolved Anisotropy Study on the Excited-State Intramolecular Proton Transfer of 1-Hydroxyanthraquinone

  • Choi, Jun-Rye;Jeoung, Sae-Chae;Cho, Dae-Won
    • Bulletin of the Korean Chemical Society
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    • v.24 no.11
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    • pp.1675-1679
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    • 2003
  • The photodynamics of excited-state intramolecular proton transfer reaction of 1-hydroxyanthraquinone (1-HAQ) and 1-deuterioanthraquinone was investigated in toluene with time-resolved emission and femtosecond transient transmittance techniques at room temperature. The temporal profiles of transient transmittance of 1-HAQ could be well described with multi-decaying time constants. The ultrafast time constant within ca. 260 fs reflects the dynamics of proton transfer. The decay component of 2 ps is assigned to an additional proton translocation process induced by the intramolecular vibrational relaxation, whereas the decay component of 18 ps is assigned to the vibrational cooling process, while the long component (200 ps) can be explained in terms of the relaxation from excited-state keto-tautomer to its ground state. Time-resolved anisotropy decay dynamics and isotope effects on the photodynamics reveals that the ESIPT from enol-tautomer to keto-one of 1-HAQ is barrierless reaction and coupled to a vibrational relaxation process.

Calcium Signaling-mediated and Differential Induction of Calmodulin Gene Expression by Stress in Oryza sativa L.

  • Phean-o-pas, Srivilai;Punteeranurak, Pornpimon;Buaboocha, Teerapong
    • BMB Reports
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    • v.38 no.4
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    • pp.432-439
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    • 2005
  • $Ca^{2+}$/calmodulin transduction pathways have been implicated in mediating stress response and tolerance in plants. Here, three genes encoding calmodulin (Cam) members of the EF-hand family of $Ca^{2+}$-binding proteins were identified from Oryza sativa L. databases. Complementary DNA for each of the calmodulin genes, OsCam1, OsCam2, and OsCam3 were sequenced. OsCam1 and OsCam2 encode a conventional 148-amino acid calmodulin protein that contains four characteristic $Ca^{2+}$-binding motifs. OsCam3 encode a similar protein with a 38-amino-acid extension containing a putative prenylation site (CVIL) at the carboxyl terminus. RT-PCR showed that each of the genes is expressed in leaves and roots of 2-week old rice seedlings. By RNA gel blot analysis, OsCam1 mRNA levels strongly increased in response to NaCl, mannitol and wounding treatments. In contrast, OsCam2 mRNA levels were relatively unchanged under all conditions investigated. NaCl treatment and wounding also increased the OsCam3 mRNA level, but in a more transient manner. Our results indicate that although the expression of genes encoding different calmodulin isoforms is ubiquitous, they are differentially regulated by various stress signals. In addition, we have demonstrated that the calcium-channel blocker lanthanum chloride inhibited the induction of OsCam1 gene expression by both NaCl and mannitol treatments. These results suggest that osmotic stress induced expression of OsCam1 gene requires the $[Ca^{2+}]_{cyt}$ elevation that is known to occur in response to these stimuli.