• Title/Summary/Keyword: Ca transient

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Intragenic Control of Expression of a Rice MADS Box Gene OsMADS1

  • Jeon, Jong-Seong;Lee, Sichul;An, Gynheung
    • Molecules and Cells
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    • v.26 no.5
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    • pp.474-480
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    • 2008
  • OsMADS1 is a rice MADS box gene necessary for floral development. To identify the key cis-regulatory regions for its expression, we utilized transgenic rice plants expressing GUS fusion constructs. Histochemical analysis revealed that the 5.7-kb OsMADS1 intragenic sequences, encompassing exon 1, intron 1, and a part of exon 2, together with the 1.9-kb 5' upstream promoter region, are required for the GUS expression pattern that coincides with flower-preferential expression of OsMADS1. In contrast, the 5' upstream promoter sequence lacking this intragenic region caused ectopic expression of the reporter gene in both vegetative and reproductive tissues. Notably, incorporation of the intragenic region into the CaMV35S promoter directed the GUS expression pattern similar to that of the endogenous spatial expression of OsMADS1 in flowers. In addition, our transient gene expression assay revealed that the large first intron following the CaMV35S minimal promoter enhances flower-preferential expression of GUS. These results suggest that the OsMADS1 intragenic sequence, largely intron 1, contains a key regulatory region(s) essential for expression.

Physiological functions of the TRPM4 channels via protein interactions

  • Cho, Chang-Hoon;Lee, Young-Sun;Kim, Eunju;Hwang, Eun Mi;Park, Jae-Yong
    • BMB Reports
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    • v.48 no.1
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    • pp.1-5
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    • 2015
  • Transient Receptor Potential, Melastatin-related, member 4 (TRPM4) channels are $Ca^{2+}$-activated $Ca^{2+}$-impermeable cation channels. These channels are expressed in various types of mammalian tissues including the brain and are implicated in many diverse physiological and pathophysiological conditions. In the past several years, the trafficking processes and regulatory mechanism of these channels and their interacting proteins have been uncovered. Here in this minireview, we summarize the current understanding of the trafficking mechanism of TRPM4 channels on the plasma membrane as well as heteromeric complex formation via protein interactions. We also describe physiological implications of protein-TRPM4 interactions and suggest TRPM4 channels as therapeutic targets in many related diseases.

FISS and SDO Observation of a Brightening Event Near a Pore

  • Kang, Juhyeong;Chae, Jongchul
    • The Bulletin of The Korean Astronomical Society
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    • v.42 no.2
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    • pp.61.2-61.2
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    • 2017
  • We report a fine scale transient brightening event near a pore boundary with the Fast Imaging Solar Spectrograph (FISS) of the 1.6m Goode Solar Telescope (GST), the Atmospheric Imaging Assembly (AIA) aboard the Solar Dynamics Observatory (SDO), and Helioseismic and Magnetic Imager (HMI) aboard SDO. The event appears in all AIA extreme ultraviolet bands, also in the two FISS lines, $H{\alpha}$ and Ca II $8542{\AA}$, and lasted for a minute. The brightening occurred at a footpoint of a loop. The conjugate brightening occurred at the other foot point outside the FISS field of view. The brightening near the pore exhibit a redshift of 4.3 km s-1 in the $H{\alpha}$ and about 2.3 km s-1 in Ca II line. Differential emission measure derived from 6 AIA EUV passbands and cloud model fitting of the two FISS lines indicate the temperature increase of between 10,000 and 20 MK at the main event. After the brightening, the upward mass motion appears in the AIA images. We discuss the physical implication of this brightening in the context of magnetic reconnection and coronal heating.

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SI Engine Closed-loop Spark Advance Control Using Cylinder Pressure (실린더 압력을 이용한 SI엔진의 페루프 점화시기 제어에 관한 연구)

  • Park, Seung-Beom;Yun, Pal-Ju
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.24 no.9 s.180
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    • pp.2361-2370
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    • 2000
  • The introduction of inexpensive cylinder pressure sensors provides new opportunities for precise engine control. This paper presents a control strategy of spark advance based upon cylinder pressure of spark ignition engines. A location of peak pressure(LPP) is the major parameter for controlling the spark timing, and also the UP is estimated, using a multi-layer feedforward neural network, which needs only five pressure sensor output voltage samples at -40˚, -20˚, 0˚, 20˚, 40˚ after top dead center. The neural network plays an important role in mitigating the A/D conversion load of an electronic engine controller by increasing the sampling interval from 10 crank angle(CA) to 20˚ CA. A proposed control algorithm does not need a sensor calibration and pegging(bias calculation) procedure because the neural network estimates the UP from the raw sensor output voltage. The estimated LPP can be regarded as a good index for combustion phasing, and can also be used as an MBT control parameter. The feasibility of this methodology is closely examined through steady and transient engine operations to control individual cylinder spark advance. The experimental results have revealed a favorable agreement of individual cylinder optimal combustion phasing.

Tissue Specific Expression of Wound-Inducible RCaM-2 Promoter in Transgenic Tobacco Plants (상처에 의해서 유도되는 벼 calmodulin promoter의 transgenic 담배에서조직 특이적 발현)

  • Choi Young Ju
    • Journal of Life Science
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    • v.15 no.2 s.69
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    • pp.176-181
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    • 2005
  • To study calmodulin (CaM) gene expression and its regulation, rice CaM promoter (RCaM-2) was isolated and fused to $\beta-glucuronidase$ (GUS), reporter gene. X-Glue staining patterns revealed that GUS localization is high in meristemic tissues such as the stem apex, stolen tip, and vascular regions. GUS staining in the transverse sections of stem and petiole was restricted to the inside of the vascular system, and cortex and epidermis located outside of the vascular system usually did not show GUS staining even a plant that expressed strong activity. GUS activity was found to be tissue specific expressed and exhibited a dramatic transient increase in response to wounding. These results suggest that the 5'-flanking region of RCaM gene regulates wound-inducible expression.

Comparative In Vitro Toxicity Study of Docetaxel and Nanoxel, a Docetaxel-Loaded Micellar Formulation Using Cultured and Blood Cells

  • Do, Van Quan;Park, Kwang-Hoon;Park, Jung-Min;Lee, Moo-Yeol
    • Toxicological Research
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    • v.35 no.2
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    • pp.201-207
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    • 2019
  • Nanoxel-$PM^{TM}$ (Nanoxel) is a docetaxel-loaded methoxy-poly(ethylene glycol)-block-poly(D,L-lactide) (mPEG-PDLLA). This newly developed and marketed nanoformulation exhibits an improved pharmacokinetic profile, efficacy, and safety. Although the safety of Nanoxel to docetaxel as well as its bioequivalence must be clinically confirmed, all biological activities have not been examined in in vitro or in vivo studies. Here, the toxicity in a cultured cell system and the effects on blood cells were tested with Nanoxel and docetaxel. The in vitro cytotoxicity of Nanoxel was found to be comparable to or slightly lower than that of docetaxel depending on the concentrations tested or the cell types. Neither docetaxel nor Nanoxel induced erythrocytes hemolysis and produced reactive oxygen species up to $100{\mu}M$. However, Nanoxel was able to enhance the aggregatory response of platelets to collagen, whereas docetaxel attenuated such aggregation in a range of $50-100{\mu}M$, while thrombin-induced aggregation was not affected by either of them. Docetaxel or Nanoxel did not alter basal level of $Ca^{2+}$ and 5-hydroxytryptamine-evoked $Ca^{2+}$ transient in vascular smooth muscle cells. These results suggest that the mPEG-PDLLA micellar formulation alters the toxicological properties of docetaxel, and that extra cautions are needed when evaluating the safety of nanomedicine.

T-Type Calcium Channels Are Required to Maintain Viability of Neural Progenitor Cells

  • Kim, Ji-Woon;Oh, Hyun Ah;Lee, Sung Hoon;Kim, Ki Chan;Eun, Pyung Hwa;Ko, Mee Jung;Gonzales, Edson Luck T.;Seung, Hana;Kim, Seonmin;Bahn, Geon Ho;Shin, Chan Young
    • Biomolecules & Therapeutics
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    • v.26 no.5
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    • pp.439-445
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    • 2018
  • T-type calcium channels are low voltage-activated calcium channels that evoke small and transient calcium currents. Recently, T-type calcium channels have been implicated in neurodevelopmental disorders such as autism spectrum disorder and neural tube defects. However, their function during embryonic development is largely unknown. Here, we investigated the function and expression of T-type calcium channels in embryonic neural progenitor cells (NPCs). First, we compared the expression of T-type calcium channel subtypes (CaV3.1, 3.2, and 3.3) in NPCs and differentiated neural cells (neurons and astrocytes). We detected all subtypes in neurons but not in astrocytes. In NPCs, CaV3.1 was the dominant subtype, whereas CaV3.2 was weakly expressed, and CaV3.3 was not detected. Next, we determined CaV3.1 expression levels in the cortex during early brain development. Expression levels of CaV3.1 in the embryonic period were transiently decreased during the perinatal period and increased at postnatal day 11. We then pharmacologically blocked T-type calcium channels to determine the effects in neuronal cells. The blockade of T-type calcium channels reduced cell viability, and induced apoptotic cell death in NPCs but not in differentiated astrocytes. Furthermore, blocking T-type calcium channels rapidly reduced AKT-phosphorylation (Ser473) and $GSK3{\beta}$-phosphorylation (Ser9). Our results suggest that T-type calcium channels play essential roles in maintaining NPC viability, and T-type calcium channel blockers are toxic to embryonic neural cells, and may potentially be responsible for neurodevelopmental disorders.

TRPV1 in Salivary Gland Epithelial Cells Is Not Involved in Salivary Secretion via Transcellular Pathway

  • Choi, Seulki;Shin, Yong-Hwan;Namkoong, Eun;Hwang, Sung-Min;Cong, Xin;Yu, Guangyan;Park, Kyungpyo
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.6
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    • pp.525-530
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    • 2014
  • Transient receptor potential vanilloid subtype 1 (TRPV1) was originally found in sensory neurons. Recently, it has been reported that TRPV1 is expressed in salivary gland epithelial cells (SGEC). However, the physiological role of TRPV1 in salivary secretion remains to be elucidated. We found that TRPV1 is expressed in mouse and human submandibular glands (SMG) and HSG cells, originated from human submandibular gland ducts at both mRNA and protein levels. However, capsaicin (CAP), TRPV1 agonist, had little effect on intracellular free calcium concentration ($[Ca^{2+}]_i$) in these cells, although carbachol consistently increased $[Ca^{2+}]_i$. Exposure of cells to high temperature (> $43^{\circ}C$) or acidic bath solution (pH5.4) did not increase $[Ca^{2+}]_i$, either. We further examined the role of TRPV1 in salivary secretion using TRPV1 knock-out mice. There was no significant difference in the pilocarpine (PILO)-induced salivary flow rate between wild-type and TRPV1 knock-out mice. Saliva flow rate also showed insignificant change in the mice treated with PILO plus CAP compared with that in mice treated with PILO alone. Taken together, our results suggest that although TRPV1 is expressed in SGEC, it appears not to play any direct roles in saliva secretion via transcellular pathway.

Dose-Dependent Cytotoxic Effects of Menthol on Human Malignant Melanoma A-375 Cells: Correlation with TRPM8 Transcript Expression

  • Kijpornyongpan, Teeratas;Sereemaspun, Amornpun;Chanchao, Chanpen
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.4
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    • pp.1551-1556
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    • 2014
  • Background: Transient receptor potential melastatin 8 (TRPM8), a principle membrane receptor involved in calcium ion influx and cell signal transduction, has been found to be up-regulated in some cancer types, including melanomas. Efficiency of menthol, an agonist of TRPM8, in killing melanoma cancer cells has been reported previously, but the mechanisms remain unclear. We here determined whether in vitro cytotoxic effects of menthol on A-375 human malignant melanoma cells might be related to TRPM8 transcript expression. Materials and Methods: The $PrestoBlue^{(R)}$ cell viability assay was used to assess the in vitro cytotoxic effect of menthol after 24h of treatment. RT-PCR was used to quantify TRPM8 transcript expression levels in normal and menthol-treated cells. Cell morphology was observed under inverted phase contrast light microscopy. Results: TRPM8 transcript expression was found at low levels in A-375 cells and down-regulated in a potentially dose-dependent manner by menthol. Menthol exerted in vitro cytotoxic effects on A-375 cells with an $IC_{50}$ value of 11.8 ${\mu}M$, which was at least as effective as 5-fluorouracil ($IC_{50}=120{\mu}M$), a commonly applied chemotherapeutic drug. Menthol showed no dose-dependent cytotoxicity on HeLa cells, a TRPM8 non-expressing cell line. Conclusions: The cytotoxic effects on A-375 cells caused by menthol might be related to reduction of the TRPM8 transcript level. This suggests that menthol might activate TRPM8 to increase cytosolic $Ca^{2+}$ levels, which leads to cytosolic $Ca^{2+}$ imbalance and triggers cell death.

The Action and Pacemaker Potential in the Frog Truncus Arteriosus (개구리 대동맥의 활동전압 및 Pacemaker 전압에 관한 연구)

  • Earm, Yung-E;Sung, Ho-Kyung
    • The Korean Journal of Physiology
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    • v.16 no.2
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    • pp.119-128
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    • 1982
  • The frog truncus arterious were studied with conventional glass microelectrode technique in order to elucidate the underlying mechanism of spontaneous pacemaker activity. The analyses were focussed on the ionic nature of pacemaker current by changing the concentrations of extracellular $K^+$ and, $Na^+$, or by using blockers of K- and Ca-current and chronotropic transmitters. 1) The action potential of the spontaneously active truncus arteriosus has some characteristic feature of maximal distolic potential ranged from -65 to -75 mV, resting potential from -45 to -50 mV and overshoot voltage about +30 mV, respectively. Duration of the action potential taken from rapid upstroke to maximal diastolic potential was about 600 msec. Usual discharge rate was $25{\sim}30/min$ at room temperature $(18{\sim}20^{\circ}C)$. 2) The sensitivity of the resting membrane potential to change extracellular potassium concentrations $(0{\sim}12\;mM)$ was relatively low. Transient hyperpolarization was appeared in the 12 mM K Ringer after 10 min exposure to 0 mM K and it could be related to Na-pump reactivation by high potassium. 3) Reduction of extracellular sodium concetrations diminished the amplitude and frequency of the action potential. In Ringer solution containing 30% Na (substituted by equimolar Tris), spontaneous activity stopped but reappeared as very slow and small action potential. There was no spotaneous activity in zero Na Ringer solution. 4) Caesium(10 mM), K-current blocker decreased the frequency of the action potential and also pacemaker depolarization. Manganese (2 mM) known to be Ca-current antagonist, blocked spontaneous activity completely. 5) Adrenaline and acetylcholine had no chronotropic effect. But adrenaline increased the duration of plateau phase and the magnitude of the action potential in the follower cell. It is concluded that K-, Na-and Ca-current components are involved in the genesis of spontaneous activity of the frog truncus arteriosus like cardiac pacemaker tissues. But the insensitivity of truncus arteriosus to adrenaline and acetylcholine indicates that there are some different control mechanisms of spontaneous rhythm in two tissues.

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