• Animal cells and systems
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• 제15권2호
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• pp.131-138
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• 2011

Redox regulation is one of the ubiquitous mechanisms to modulate ion channels. We here investigated how 5,5'-dithio-bis (2-nitrobenzoic acid), a cysteine specific oxidizing reagent, modulates $Ca_v3.1$ and $Ca_v3.2$ T-type $Ca^{2+}$ channels expressed in Xenopus oocytes. Application of the reagent inhibited $Ca_v3.1$ and $Ca_v3.2$ currents in a dose-dependent manner. The oxidizing reagent (1 mM) reduced the peak amplitude of $Ca_v3.1$ and $Ca_v3.2$ currents by ~50% over 2-3 minutes and the decreased currents were fully recovered upon washout of it. The reagent slowed the activation and inactivation kinetics of $Ca_v3.1$, $Ca_v3.2$, and $Ca_v3.3$ channel currents. Notably, the reagent positively shifted both activation and steady-state inactivation curves of $Ca_v3.1$, while it did not those of $Ca_v3.2$. Utilizing chimeric channels from $Ca_v3.1$ and $Ca_v3.2$, we localized the domains III and IV of $Ca_v3.1$ responsible for the positive shifts of channel activation and steady-state inactivation. These findings provide hints relevant to the electrophysiological and molecular mechanisms accounting for the oxidative regulation of T-type channels.

• Animal cells and systems
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• 제12권3호
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• pp.137-141
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• 2008

Molecular cloning revealed the three isoforms($Ca_v3.1,\;Ca_v3.2,\;and\;Ca_v3.3$) of the T-type calcium channel subfamily. Expression studies exhibited their distinctive electrophysiological and pharmacological properties, accounting for diverse properties of T-type calcium channel currents previously characterized from isolated cells. However, electrophysiological properties of ion channels have shown to be more diversified by their splice variants. We here searched splice variants of rat $Ca_v3.1$ T-type channel by reverse-transcription-polymerase chain reaction(RT-PCR) to further explore diversity of $Ca_v3.1$. Interestingly, analyses of cloned RT-PCR products displayed that there were at least four splicing variants of rat $Ca_v3.1$ in the loop connecting repeats III and IV. Southern blot analyses indicated that the predominantly detected variant in brain was $Ca_v3.1a$(492 bp), which were rarely detected in most of peripheral tissues. Other two variants($Ca_v3.1c$, 546 bp; $Ca_v3.1d$, 525 bp) were detected in most of the tissues examined. The smallest isoform($Ca_v3.1b$, 471 bp) was rarely detected all the tissues. Electrophysiological characterization of the splicing variants indicated that the splice variants differ in inactivation kinetics and the voltage dependence of activation and inactivation as well.

• Biomolecules & Therapeutics
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• 제26권5호
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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.

• Journal of Microbiology and Biotechnology
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• 제11권4호
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• pp.614-618
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• 2001

Low-threshold T-type $Ca^{2+}$ channels are distinctive voltage-operated gates for external $Ca^{2+}$ entry around a resting membrane potential due to their low voltage activation. These phenomena have already been extensively studied due to their relevance in diverse physiological functions. Recently, three T-type $Ca^{2+}$ channel ${\alpha}$$_1$subunits were cloned and their biophysical properties were characterized after expression in mammalian expression systems. In this study, ${\alpha_IG} and {\alpha_IH}$ low-threshold $Ca^{2+}$ channels were expressed and characterized in Xenopus oocytes after adding 5' and 3'untranslated portions of a Xenopus ${\beta}$ globin to improve their expression levels. The added portions dramatically enhanced the expression levels of the ${\alpha_IG} and {\alpha_IH}$ T-type channels. When currents were recorded in 10 mM $Ba^{2+}$ as the charge carrier, the activation thresholds were about -60 mV, peak currents appeared at -20 mV, and the reversal potentials were between +40 and +45. The activation time constants were very similar to each other, while the inactivation time constants of the ${\alpha_IG}$ currents were smaller than those of ${\alpha_IH}$. Taken together, the electrophysiological properties of the ${\alpha_IG} and {\alpha_IH}$ channels expressed in Xenopus oocytes were similar to the previously reported characteristics of low-threshold $Ca^{2+}$ channel currents.

• Molecules and Cells
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• 제25권2호
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• pp.242-246
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• 2008

To assess the role of $\alpha_{1G}$ T-type $Ca^{2+}$ channels in neuropathic pain after L5 spinal nerve ligation, we examined behavioral pain susceptibility in mice lacking $Ca_{V}3.1$ (${\alpha}_{1G}{^{-/-}}$), the gene encoding the pore-forming units of these channels. Reduced spontaneous pain responses and an increased threshold for paw withdrawal in response to mechanical stimulation were observed in these mice. The ${{\alpha}_{1G}}^{-/-}$ mice also showed attenuated thermal hyperalgesia in response to both low-(IR30) and high-intensity (IR60) infrared stimulation. Our results reveal the importance of ${\alpha}_{1G}$ T-type $Ca^{2+}$ channels in the development of neuropathic pain, and suggest that selective modulation of ${\alpha}_{1G}$ subtype channels may provide a novel approach to the treatment of allodynia and hyperalgesia.

• 한국수정란이식학회지
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• 제16권2호
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• pp.79-89
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• 2001

배분화과정시 나타나는 $Ca^{2+}$ 변화에 미치는 $E_2$의 영향을 알아보고자 whole cell voltage clamp 기법, 방사선 등위원소 면역측정법, 그리고 공초점 현미경을 통하여 $E_2$처리 후 나타나는 $Ca^{2+}$ 전류 변화 및 세포내 $Ca^{2+}$ 농도 변화를 조사하였다. 생쥐의 미성숙 난자는 난소의 난포를 천자하고, 배란난자는 과배란 처리 후 난관에서 회수하였다. 수정란은 과배란 처리 후 수컷 생쥐와 교미를 유도한 후 각각의 단계에 맞는 수정란을 채란하였다. 혈중 $E_2$의 농도는 심장을 천자하여 혈액을 채취한 후 배발달 단계와 호르몬 처리 시간이 일치하는 혈액만을 사용하였다. 본 실험의 결과를 요약하면 다음과 같다. 1. $E_2$처리시 미성숙난자의 제 1극체 형성률 (성숙의 지표)은 $E_2$를 처리하지 않은 난자(83% : 83/100)보다 $E_2$를 처리한 난자 (94%, 94/100)에서 유의적 (P<0.05)으로 높게 나타났다. 2. $E_2$를 처리하였을 때 $Ca^{2+}$ 내향전류의 변화는 -10 mV에서 -1.23$\pm$0.01 nA (n=15)에서 -1.50$\pm$0.03 nA (n=15)로 122% 상승함으로써 유의한 (P<0.05) 변화를 보였다. 3. $E_2$를 처리하지 않은 난자 및 수정란을 1로 한 후 $E_2$를 처리한 난자 및 수정란의 변화를 상대적인 값으로 표시하였다. $E_2$처리한 난자는 1.22$\pm$0.17 (n=10), $E_2$처리한 전핵배는 1.20$\pm$0.14 (n=10), $E_2$처리한 2세포기배는 1.07$\pm$0.01 (n=10), 4세포기배는 1.05$\pm$0.09 (n=10)를 나타냄으로써 수정란의 단계마다 $E_2$의 반응 결과가 차이가 남을 알 수 있었다. 4. $E_2$농도 곡선에서 PMSG 처리 후 $E_2$의 혈중농도는 계속적인 상승을 보이다가 배란시기에 최고치를 나타내었으며, 배란 후 다시 감소하여 8세포기에서는 급격한 감소현상이 나타났다. 이후 다시 상실기를 거쳐 배반포기 임신기간동안 $E_2$의 농도가 상승하였다. 5. $E_2$처리 후 세포내 $Ca^{2+}$ 농도변화의 결과로, $E_2$를 처리하지 않은 난자들의 세포내 $Ca^{2+}$ 농도는 836.4$\pm$131.2 (n=10), $E_2$를 처리한 난자들은 1736.4$\pm$192.0 (n=10)로써 유의한 (P<0.05) 차이를 보였다. 이상의 결과로부터 $E_2$처리에 의한 세포내 $Ca^{2+}$ 농도 상승은 $E_2$가 $Ca^{2+}$ 통로를 자극함으로써 세포바깥의 $Ca^{2+}$이 세포안으로 이동하여 나타나는 변화로 생각된다.

• 한국농공학회지
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• 제16권1호
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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)