• The Korean Journal of Physiology and Pharmacology
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• 제23권3호
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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.

• 한국재료학회지
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• 제29권4호
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• pp.241-251
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• 2019

There are several manufacturing techniques for developing thermionic cathodes for vacuum ultraviolet(VUV) ionizers. The triple alkaline earth metal emitters(Ca-Sr-Ba) are formulated as efficient and reliable thermo-electron sources with a great many different compositions for the ionizing devices. We prepare two basic suspensions with different compositions: calcium, strontium and barium. After evaluating the electron-emitting performance for europium, gadolinium, and yttrium-based cathodes mixed with these suspensions, we selected the yttrium for its better performance. Next, another transition metal indium and a lanthanide metal neodymium salt is introduced to two base emitters. These final composite metal emitters are coated on the tungsten filament and then activated to the oxide cathodes by an intentionally programmed calcination process under an ultra-high vacuum(${\sim}10^{-6}torr$). The performance of electron emission of the cathodes is characterized by their anode currents with respect to the addition of each element, In and Nd, and their concentration of cathodes. Compared to both the base cathodes, the electron emission performance of the cathodes containing indium and neodymium decreases. The anode current of the Nd cathode is more markedly degraded than that with In.

• The Korean Journal of Physiology and Pharmacology
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• 제14권1호
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• pp.21-28
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• 2010

Phenolic compounds affect intracellular free $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) signaling. The study examined whether the simple phenolic compound octyl gallate affects ATP-induced $Ca^{2+}$ signaling in PC12 cells using fura-2-based digital $Ca^{2+}$ imaging and whole-cell patch clamping. Treatment with ATP ($100\;{\mu}M$) for 90 s induced increases in $[Ca^{2+}]_i$ in PC12 cells. Pretreatment with octyl gallate (100 nM to $20\;{\mu}M$) for 10 min inhibited the ATP-induced $[Ca^{2+}]_i$ response in a concentration-dependent manner ($IC_{50}=2.84\;{\mu}M$). Treatment with octyl gallate ($3\;{\mu}M$) for 10 min significantly inhibited the ATP-induced response following the removal of extracellular $Ca^{2+}$ with nominally $Ca^{2+}$-free HEPES HBSS or depletion of intracellular $Ca^{2+}$ stores with thapsigargin ($1\;{\mu}M$). Treatment for 10 min with the L-type $Ca^{2+}$ channel antagonist nimodipine ($1\;{\mu}M$) significantly inhibited the ATP-induced $[Ca^{2+}]_i$ increase, and treatment with octyl gallate further inhibited the ATP-induced response. Treatment with octyl gallate significantly inhibited the $[Ca^{2+}]_i$ increase induced by 50 mM KCI. Pretreatment with protein kinase C inhibitors staurosporin (100 nM) and GF109203X (300 nM), or the tyrosine kinase inhibitor genistein ($50\;{\mu}M$) did not significantly affect the inhibitory effects of octyl gallate on the ATP-induced response. Treatment with octyl gallate markedly inhibited the ATP-induced currents. Therefore, we conclude that octyl gallate inhibits ATP-induced $[Ca^{2+}]_i$ increase in PC12 cells by inhibiting both non-selective P2X receptor-mediated influx of $Ca^{2+}$ from extracellular space and P2Y receptor-induced release of $Ca^{2+}$ from intracellular stores in protein kinase-independent manner. In addition, octyl gallate inhibits the ATP-induced $Ca^{2+}$ responses by inhibiting the secondary activation of voltage-gated $Ca^{2+}$ channels.

• 대한의생명과학회지
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• 제15권3호
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• pp.199-205
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• 2009

This study was designed to investigate direct effects of [D-$Pen^2$, D-$Pen^5$]-enkephalin, a $\delta$-opioid receptor agonist on the neuronal activity of medial vestibular nuclear (MVN) neurons by whole-cell configuration patch clamp experiments. The spike frequency of MVN neuron was increased to $9.50{\pm}0.55$ (P<0.05) and $10.56{\pm}0.66$ (P<0.05) by 5 and $10{\mu}M$ [D-$Pen^2$, D-$Pen^5$]-enkephalin from the control level of $8.05{\pm}0.55$ spikes/sec, respectively (n=18). The resting membrane potential of the neurons was increased to $-37.86{\pm}0.92$ and $-36.97{\pm}0.97$ (P<0.05) from $-38.74{\pm}1.13\;mV$ by 5 and $10{\mu}M$ [D-$Pen^2$, D-$Pen^5$]-enkephalin, respectively. The amplitude of afterhyperpolarization was decreased to $23.78{\pm}0.65$ and $21.67{\pm}0.89$ (P<0.05) from $23.73{\pm}0.53\;mV$ by 5 and $10{\mu}M$ [D-$Pen^2$, D-$Pen^5$]-enkephalin, respectively. The spike width was changed to $2.22{\pm}0.08$ and $2.24{\pm}0.07$ from $2.20{\pm}0.08\;mV$ by 5 and $10{\mu}M$ [D-$Pen^2$, D-$Pen^5$]-enkephalin, respectively. After pretreatment of naltrindole, a highly selective 8-opioid receptor antagonist, [D-$Pen^2$, D-$Pen^5$]-enkephalin did not change firing rate, resting membrane potential, afterhyperpolarization amplitude, and spike width of MVN neurons. The above experimental results suggest that [D-$Pen^2$, D-$Pen^5$]-enkephalin increases the neuronal activity of MVN neurons via inhibition of calcium-dependent potassium currents underlying the afterhyperpolarization.

• Journal of Advanced Marine Engineering and Technology
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• 제39권7호
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• pp.779-785
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• 2015

일반적으로 선박 및 해양구조물, 그리고 심해 설비의 가장 적절한 부식 방지법으로는 음극방식법이 널리 사용되고 있다. 해수 중에 이러한 음극방식법이 적용될 경우, 음극전류가 용존 해 있는 산소를 환원시킴으로써 음극분극 된 설비 표면에 수산화 이온을 다량 발생시키게 되고, 이로 인하여 계면에서의 pH는 증가하게 된다. 또한 탄산이온의 농도 역시 증가하게 되어, 바닷물 속에 용해되어 있는 마그네슘과 칼슘 이온들이 이들 수산화 이온 및 탄산 이온과 결합함으로써, 수산화마그네슘 및 탄산칼슘을 주성분으로 하는 무기물 층이 설비 표면에 석출하게 되는데 이렇게 형성된 피막을 일반적으로 "석회질 피막"이라고 한다. 이러한 무기물 층은 해수라는 부식환경에서 모재를 보호하는 물리적 장벽으로서의 역할을 함은 물론 음극방식을 할 때 요구되는 전류밀도를 감소시키는 역할을 하게 된다. 한편 이러한 해수 중에서의 석회질 피막의 형성은 전위, 전류, pH, 온도, 시간 등을 포함한 많은 변수들에 의해 변화하게 되는데, 이에 본 연구에서는 여러 가지 변수들 중 특히나 해수의 온도 및 시험편 표면 조건에 따른 무기물 층의 피막구조변화 및 특성변화를 살펴봄으로서 환경친화적인 코팅막의 개발에 대한 설계지침을 제공 할 수 있었다.

• 공업화학
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• 제23권5호
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• pp.474-479
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• 2012

막결합 축전식 탈염(MCDI) 기술을 이용하여 $Na^{+}$와 $Ca^{2+}$ 이온이 혼합된 용액에서 $Ca^{2+}$ 이온의 선택적 제거 가능성을 연구하였다. 양이온교환막인 CMX막에 대한 $Ca^{2+}$ 이온의 선택성을 확인하기 위해 흡착평형 실험을 실시하였다. 그리고 MCDI 셀을 이용해 혼합용액(5 meq/L NaCl + 2 meq/L $CaCl_{2}$)에 대한 탈염실험을 수행하였다. 흡착평형 실험결과 용액과 CMX막에서 $Ca^{2+}$ 이온의 당량분율은 각각 28.6, 87.2%를 보여 CMX막이 $Ca^{2+}$ 이온에 대해 높은 선택성을 갖는 것을 확인하였다. MCDI 셀에 일정한 전류를 인가하면서 셀 전위가 1.0 V에 도달할 때까지 탈염실험을 실시하였다. 그 결과 흡착된 이온의 총량은 인가한 전류밀도에 큰 영향을 받지 않고 일정하였다. 그러나 흡착된 이온 중 $Ca^{2+}$ 이온의 비율은 전류밀도에 반비례하였으며 200, 300, 500, $700\;A/m^{2}$의 전류밀도에서 각각 81.4, 78.4, 77.0, 74.5%로 나타났다. 이러한 결과는 낮은 전류밀도에서 CMX막에 흡착된 $Ca^{2+}$ 이온의 비율이 높았기 때문인 것으로 판단된다.

• The Korean Journal of Physiology and Pharmacology
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• 제17권3호
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• pp.223-228
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• 2013

The calcium-activated $K^+$ ($BK_{Ca}$) channel is one of the potassium-selective ion channels that are present in the nervous and vascular systems. $Ca^{2+}$ is the main regulator of $BK_{Ca}$ channel activation. The $BK_{Ca}$ channel contains two high affinity $Ca^{2+}$ binding sites, namely, regulators of $K^+$ conductance, RCK1 and the $Ca^{2+}$ bowl. Lysophosphatidic acid (LPA, 1-radyl-2-hydroxy-sn-glycero-3-phosphate) is one of the neurolipids. LPA affects diverse cellular functions on many cell types through G protein-coupled LPA receptor subtypes. The activation of LPA receptors induces transient elevation of intracellular $Ca^{2+}$ levels through diverse G proteins such as $G{\alpha}_{q/11}$, $G{\alpha}_i$, $G{\alpha}_{12/13}$, and $G{\alpha}s$ and the related signal transduction pathway. In the present study, we examined LPA effects on $BK_{Ca}$ channel activity expressed in Xenopus oocytes, which are known to endogenously express the LPA receptor. Treatment with LPA induced a large outward current in a reversible and concentration-dependent manner. However, repeated treatment with LPA induced a rapid desensitization, and the LPA receptor antagonist Ki16425 blocked LPA action. LPA-mediated $BK_{Ca}$ channel activation was also attenuated by the PLC inhibitor U-73122, $IP_3$ inhibitor 2-APB, $Ca^{2+}$ chelator BAPTA, or PKC inhibitor calphostin. In addition, mutations in RCK1 and RCK2 also attenuated LPA-mediated $BK_{Ca}$ channel activation. The present study indicates that LPA-mediated activation of the $BK_{Ca}$ channel is achieved through the PLC, $IP_3$, $Ca^{2+}$, and PKC pathway and that LPA-mediated activation of the $BK_{Ca}$ channel could be one of the biological effects of LPA in the nervous and vascular systems.

• 한국수정란이식학회지
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• 제21권1호
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• pp.35-43
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• 2006

이온 통로 및 이온 농도의 변화는 수정 현상을 포함한 다양한 세포 기능에 중요한 역할을 한다. 그러나 이러한 이온의 변화가 포유동물 배의 발달과정에 어떻게 관여하는지에 대해서는 알려진 바가 적다. 본 연구에서는 생쥐난자가 수정 이후 배 발달 과정을 거치는 동안 나타나는 칼슘과 포타슘 이온의 변화를 전기생리학적 실험 기법과 공초점 현미경을 이용하여 조사하였다. 수정 시에 나타나는 일시적인 세포내 칼슘 농도 변화는 활성 전류(수정 전류)와 함께 동반되었다. 그러나 수정과 같은 극적인 현상이나 자극이 없는 시기에는 세포내 칼슘 농도가 배 발달 시기와 상관없이 일정한 수준으로 유지되었다. 이것은 세포내외의 칼슘 농도의 보상현상으로도 설명할 수 있을 것이다. 배 발달이 진행됨에 따라 난관액의 포타슘 농도는 계속 증가하여 8세포기 배에서는 난자보다 26% 증가하였다. 상실배, 포배기에서는 포타슘 농도가 감소하였다. 배 발달이 진행됨에 따라 주로 포타슘 이온에 의해 조절되는 막 전압은 탈분극되고, 칼슘 이온의 세포 안으로의 유입은 점점 감소하였다. 생쥐 난자에 5 mM의 칼슘을 처리하였을 때 막 전압은 일시적인 과분극 현상을 보이다가 회복되었다. 칼슘 유입에 따른 막 전압 변화에 관여하는 포타슘 통로를 확인하기 위하여 포타슘 통로 차단제를 전 처리한 후 칼슘을 처리한 결과, 칼슘만을 단독으로 처리한 결과와 유의한 차이를 보이지 않았다. 막 전압의 과분극 현상은 잘 알려진 포타슘 통로 차단제인 TEA에 억제되지 않았다. 그리고 small conductance $Ca^{2+}$-activated 포타슘 통로 차단제 인 apamin에 의해서도 억제되지 않았다. 따라서 생쥐 난자에서 과분극을 유발시키는 포타슘 통로는 TEA와 apamin에 억제되지 않는 다른 포타슘 통로로 생각된다. 이상의 결과로부터 배 발달 동안 변화되는 칼슘과 포타슘 이온은 수정 및 초기 배 발달에 중요한 인자로써 작용할 것으로 생각되며, two-pore domain 포타슘 통로가 난자의 막 전압 조절에 관여할 가능성을 제시한다.