• Title/Summary/Keyword: %24Cl%5E-%24 channel

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Acidic pH-activated $Cl^-$ Current and Intracellular $Ca^{2+}$ Response in Human Keratinocytes

  • Park, Su-Jung;Choi, Won-Woo;Kwon, Oh-Sang;Chung, Jin-Ho;Eun, Hee-Chul;Earm, Young-E;Kim, Sung-Joon
    • The Korean Journal of Physiology and Pharmacology
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    • v.12 no.4
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    • pp.177-183
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    • 2008
  • The layers of keratinocytes form an acid mantle on the surface of the skin. Herein, we investigated the effects of acidic pH on the membrane current and $[Ca^{2+}]_c$ of human primary keratinocytes from foreskins and human keratinocyte cell line (HaCaT). Acidic extracellular pH ($pH_e{\leq}5.5$) activated outwardly rectifying $Cl^-$ current ($I_{Cl,pH}$) with slow kinetics of voltage-dependent activation. $I_{Cl,pH}$ was potently inhibited by an anion channel blocker 4,4'-diisothiocyanostilbene-2,2'-disulphonic acid (DIDS, 73.5% inhibition at 1${\mu}$M). $I_{Cl,pH}$ became more sensitive to $pH_e$ by raising temperature from $24^{circ}C$ to $37^{circ}C$. HaCaT cells also expressed $Ca^{2+}$-activated $Cl^-$ current ($I_{Cl,Ca}$), and the amplitude of $I_{Cl,Ca}$ was increased by relatively weak acidic $pH_e$ (7.0 and 6.8). Interestingly, the acidic $pH_e$ (5.0) also induced a sharp increase in the intracellular [$Ca^{2+}$] (${\triangle}[Ca^{2+}]_{acid}$) of HaCaT cells. The ${\triangle}[Ca^{2+}]_{acid}$ was independent of extracellular $Ca^{2+}$, and was abolished by the pretreatment with PLC inhibitor, U73122. In primary human keratinocytes, 5 out of 28 tested cells showed ${\triangle}[Ca^{2+}]_{acid}$. In summary, we found $I_{Cl,pH}$ and ${\triangle}[Ca^{2+}]_{acid}$ in human keratinocytes, and these ionic signals might have implication in pathophysiological responses and differentiation of epidermal keratinocytes.

Properties of Single $K^{+}$ Channels of Skeletal Muscle Incorporated into Planar Lipid Bilayer

  • Park, Jin-Bong;Kim, Hee-Jeong;Cho, Myung-Haing;Lee, Hang;Park, Hong-Ki;Lee, Mun-Han;Ryu, Pan-Dong
    • The Korean Journal of Physiology
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    • v.29 no.1
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    • pp.13-27
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    • 1995
  • single $K^{+}$ channels of skeletal muscle from the rat and frog were into planar lipid bilayers and their properties were studied. Fusion was induced by an osmotic gradient. Of the four types of $K^{+}$ channels recorded, the two most frequently observed were a voltage and $Ca^{2+}-activated$ $K^{+}$ channel and a $K^{+}$ channel with a prominent conductance substate. The first $K^{+}$ channel was identified as the large $Ca^{2+}-activated$ $K^{+}$ (BK) channel because the open-state probability was increased with depolarization (e-fold change per $10.6{\pm}3.5$ mV, n=8) and internal $Ca^{2+}$ (half-activation at $16.7{\pm}3.8$ mV, n=8, pCa 4) and its conductance was large ($247{\pm}4.9$ pS, n=24 in 0.1 M KCI). Lifetime distributions of open- and closed-states could be fitted with single exponentials of several milliseconds. The mean open- and closed-lifetimes were linearly dependent on the intracellular $[Ca^{2+}]$ and $1/[Ca^{2+}]$, respectively. The second $K^{+}$ channel showed a conductance substate at $30{\sim}60%$ of the open state. Its current-voltage relation was linear in the range of $-80\;{\sim}\;+80\;mV$. The slope conductance of the substate and open-state were 40 and 144 pS in 0.2 M KCl, respectively. The channel was highly selective for $K^{+}$ over Cl. The open-state probability was weakly voltage-dependent (e-fold change per 35 mV. The lifetime distributions of open- and closed-states were fitted with two exponentials and the major gating occurred slowly at several hundred milliseconds. Based on the above results, we think the second type of $K^{+}$ channel is the sarcoplasmic reticulum $K^{+}$ (SRK) channel. In addition, both types of channel were also incorporated into the lipids extracted from the skeletal muscle. The channel properties recorded in the bilayers termed from synthetic and extracted lipids were qualitatively similar. Our data indicate that BK and SRK channels are rich in the skeletal muscle and their properties and regulation could be effectively studied in planar lipid bilayer.

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Three sesquiterpene lactones suppress lung adenocarcinoma by blocking TMEM16A-mediated Ca2+-activated Cl- channels

  • Ruilian Xiu;Jie Jia;Qing Zhang;Fengjiao Liu;Yaxin Jia;Yuanyuan Zhang;Beibei Song;Xiaodan Liu;Jingwei Chen;Dongyang Huang;Fan Zhang;Juanjuan Ma;Honglin Li;Xuan Zhang;Yunyun Geng
    • The Korean Journal of Physiology and Pharmacology
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    • v.27 no.6
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    • pp.521-531
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    • 2023
  • Transmembrane protein TMEM16A, which encodes calcium-activated chloride channel has been implicated in tumorigenesis. Overexpression of TMEM16A is associated with poor prognosis and low overall survival in multiple cancers including lung adenocarcinoma, making it a promising biomarker and therapeutic target. In this study, three structure-related sesquiterpene lactones (mecheliolide, costunolide and dehydrocostus lactone) were extracted from the traditional Chinese medicine Aucklandiae Radix and identified as novel TMEM16A inhibitors with comparable inhibitory effects. Their effects on the proliferation and migration of lung adenocarcinoma cells were examined. Whole-cell patch clamp experiments showed that these sesquiterpene lactones potently inhibited recombinant TMEM16A currents in a concentration-dependent manner. The half-maximal concentration (IC50) values for three tested sesquiterpene lactones were 29.9 ± 1.1 µM, 19.7 ± 0.4 µM, and 24.5 ± 2.1 µM, while the maximal effect (Emax) values were 100.0% ± 2.8%, 85.8% ± 0.9%, and 88.3% ± 4.6%, respectively. These sesquiterpene lactones also significantly inhibited the endogenous TMEM16A currents and proliferation, and migration of LA795 lung cancer cells. These results demonstrate that mecheliolide, costunolide and dehydrocostus lactone are novel TMEM16A inhibitors and potential candidates for lung adenocarcinoma therapy.

Studies on the Rice Yield Decreased by Ground Water Irrigation and Its Preventive Methods (지하수 관개에 의한 수도의 멸준양상과 그 방지책에 관한 연구)

  • 한욱동
    • Magazine of the Korean Society of Agricultural Engineers
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    • v.16 no.1
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    • pp.3225-3262
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    • 1974
  • The purposes of this thesis are to clarify experimentally the variation of ground water temperature in tube wells during the irrigation period of paddy rice, and the effect of ground water irrigation on the growth, grain yield and yield components of the rice plant, and, furthermore, when and why the plant is most liable to be damaged by ground water, and also to find out the effective ground water irrigation methods. The results obtained in this experiment are as follows; 1. The temperature of ground water in tube wells varies according to the location, year, and the depth of the well. The average temperatures of ground water in a tubewells, 6.3m, 8.0m deep are $14.5^{\circ}C$ and $13.1^{\circ}C$, respercively, during the irrigation period of paddy rice (From the middle of June to the end of September). In the former the temperature rises continuously from $12.3^{\circ}C$ to 16.4$^{\circ}C$ and in the latter from $12.4^{\circ}C$ to $13.8^{\circ}C$ during the same period. These temperatures are approximately the same value as the estimated temperatures. The temperature difference between the ground water and the surface water is approximately $11^{\circ}C$. 2. The results obtained from the analysis of the water quality of the "Seoho" reservoir and that of water from the tube well show that the pH values of the ground water and the surface water are 6.35 and 6.00, respectively, and inorganic components such as N, PO4, Na, Cl, SiO2 and Ca are contained more in the ground water than in the surface water while K, SO4, Fe and Mg are contained less in the ground water. 3. The response of growth, yield and yield components of paddy rice to ground water irrigation are as follows; (l) Using ground water irrigation during the watered rice nursery period(seeding date: 30 April, 1970), the chracteristics of a young rice plant, such as plant height, number of leaves, and number of tillers are inferior to those of young rice plants irrigated with surface water during the same period. (2) In cases where ground water and surface water are supplied separately by the gravity flow method, it is found that ground water irrigation to the rice plant delays the stage at which there is a maximum increase in the number of tillers by 6 days. (3) At the tillering stage of rice plant just after transplanting, the effect of ground water irrigation on the increase in the number of tillers is better, compared with the method of supplying surface water throughout the whole irrigation period. Conversely, the number of tillers is decreased by ground water irrigation at the reproductive stage. Plant height is extremely restrained by ground water irrigation. (4) Heading date is clearly delayed by the ground water irrigation when it is practised during the growth stages or at the reproductive stage only. (5) The heading date of rice plants is slightly delayed by irrigation with the gravity flow method as compared with the standing water method. (6) The response of yield and of yield components of rice to ground water irrigation are as follows: \circled1 When ground water irrigation is practised during the growth stages and the reproductive stage, the culm length of the rice plant is reduced by 11 percent and 8 percent, respectively, when compared with the surface water irrigation used throughout all the growth stages. \circled2 Panicle length is found to be the longest on the test plot in which ground water irrigation is practised at the tillering stage. A similar tendency as that seen in the culm length is observed on other test plots. \circled3 The number of panicles is found to be the least on the plot in which ground water irrigation is practised by the gravity flow method throughout all the growth stages of the rice plant. No significant difference is found between the other plots. \circled4 The number of spikelets per panicle at the various stages of rice growth at which_ surface or ground water is supplied by gravity flow method are as follows; surface water at all growth stages‥‥‥‥‥ 98.5. Ground water at all growth stages‥‥‥‥‥‥62.2 Ground water at the tillering stage‥‥‥‥‥ 82.6. Ground water at the reproductive stage ‥‥‥‥‥ 74.1. \circled5 Ripening percentage is about 70 percent on the test plot in which ground water irrigation is practised during all the growth stages and at the tillering stage only. However, when ground water irrigation is practised, at the reproductive stage, the ripening percentage is reduced to 50 percent. This means that 20 percent reduction in the ripening percentage by using ground water irrigation at the reproductive stage. \circled6 The weight of 1,000 kernels is found to show a similar tendency as in the case of ripening percentage i. e. the ground water irrigation during all the growth stages and at the reproductive stage results in a decreased weight of the 1,000 kernels. \circled7 The yield of brown rice from the various treatments are as follows; Gravity flow; Surface water at all growth stages‥‥‥‥‥‥514kg/10a. Ground water at all growth stages‥‥‥‥‥‥428kg/10a. Ground water at the reproductive stage‥‥‥‥‥‥430kg/10a. Standing water; Surface water at all growh stages‥‥‥‥‥‥556kg/10a. Ground water at all growth stages‥‥‥‥‥‥441kg/10a. Ground water at the reproductive stage‥‥‥‥‥‥450kg/10a. The above figures show that ground water irrigation by the gravity flow and by the standing water method during all the growth stages resulted in an 18 percent and a 21 percent decrease in the yield of brown rice, respectively, when compared with surface water irrigation. Also ground water irrigation by gravity flow and by standing water resulted in respective decreases in yield of 16 percent and 19 percent, compared with the surface irrigation method. 4. Results obtained from the experiments on the improvement of ground water irrigation efficiency to paddy rice are as follows; (1) When the standing water irrigation with surface water is practised, the daily average water temperature in a paddy field is 25.2$^{\circ}C$, but, when the gravity flow method is practised with the same irrigation water, the daily average water temperature is 24.5$^{\circ}C$. This means that the former is 0.7$^{\circ}C$ higher than the latter. On the other hand, when ground water is used, the daily water temperatures in a paddy field are respectively 21.$0^{\circ}C$ and 19.3$^{\circ}C$ by practising standing water and the gravity flow method. It can be seen that the former is approximately 1.$0^{\circ}C$ higher than the latter. (2) When the non-water-logged cultivation is practised, the yield of brown rice is 516.3kg/10a, while the yield of brown rice from ground water irrigation plot throughout the whole irrigation period and surface water irrigation plot are 446.3kg/10a and 556.4kg/10a, respectivelely. This means that there is no significant difference in yields between surface water irrigation practice and non-water-logged cultivation, and also means that non-water-logged cultivation results in a 12.6 percent increase in yield compared with the yield from the ground water irrigation plot. (3) The black and white coloring on the inside surface of the water warming ponds has no substantial effect on the temperature of the water. The average daily water temperatures of the various water warming ponds, having different depths, are expressed as Y=aX+b, while the daily average water temperatures at various depths in a water warming pond are expressed as Y=a(b)x (where Y: the daily average water temperature, a,b: constants depending on the type of water warming pond, X; water depth). As the depth of water warning pond is increased, the diurnal difference of the highest and the lowest water temperature is decreased, and also, the time at which the highest water temperature occurs, is delayed. (4) The degree of warming by using a polyethylene tube, 100m in length and 10cm in diameter, is 4~9$^{\circ}C$. Heat exchange rate of a polyethylene tube is 1.5 times higher than that or a water warming channel. The following equation expresses the water warming mechanism of a polyethylene tube where distance from the tube inlet, time in day and several climatic factors are given: {{{{ theta omega (dwt)= { a}_{0 } (1-e- { x} over { PHI v })+ { 2} atop { SUM from { { n}=1} { { a}_{n } } over { SQRT { 1+ {( n omega PHI) }^{2 } } } } LEFT { sin(n omega t+ { b}_{n }+ { tan}^{-1 }n omega PHI )-e- { x} over { PHI v }sin(n omega LEFT ( t- { x} over {v } RIGHT ) + { b}_{n }+ { tan}^{-1 }n omega PHI ) RIGHT } +e- { x} over { PHI v } theta i}}}}{{{{ { theta }_{$\infty$ }(t)= { { alpha theta }_{a }+ { theta }_{ w'} +(S- { B}_{s } ) { U}_{w } } over { beta } , PHI = { { cpDU}_{ omega } } over {4 beta } }}}} where $\theta$$\omega$; discharged water temperature($^{\circ}C$) $\theta$a; air temperature ($^{\circ}C$) $\theta$$\omega$';ponded water temperature($^{\circ}C$) s ; net solar radiation(ly/min) t ; time(tadian) x; tube length(cm) D; diameter(cm) ao,an,bn;constants determined from $\theta$$\omega$(t) varitation. cp; heat capacity of water(cal/$^{\circ}C$ ㎥) U,Ua; overall heat transfer coefficient(cal/$^{\circ}C$ $\textrm{cm}^2$ min-1) $\omega$;1 velocity of water in a polyethylene tube(cm/min) Bs ; heat exchange rate between water and soil(ly/min)

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