• The Korean Journal of Physiology and Pharmacology
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• v.25 no.5
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• pp.449-457
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• 2021

The sleep-wake cycle is regulated by the alternating activity of sleep- and wake-promoting neurons. The dorsal raphe nucleus (DRN) secretes 5-hydroxytryptamine (5-HT, serotonin), promoting wakefulness. Melatonin secreted from the pineal gland also promotes wakefulness in rats. Our laboratory recently demonstrated that daily changes in nitric oxide (NO) production regulates a signaling pathway involving with-no-lysine kinase (WNK), Ste20-related proline alanine rich kinase (SPAK)/oxidative stress response kinase 1 (OSR1), and cation-chloride co-transporters (CCC) in rat DRN serotonergic neurons. This study was designed to investigate the effect of melatonin on NO-regulated WNK-SPAK/OSR1-CCC signaling in wake-inducing DRN neurons to elucidate the mechanism underlying melatonin's wake-promoting actions in rats. Ex vivo treatment of DRN slices with melatonin suppressed neuronal nitric oxide synthase (nNOS) expression and increased WNK4 expression without altering WNK1, 2, or 3. Melatonin increased phosphorylation of OSR1 and the expression of sodium-potassium-chloride co-transporter 1 (NKCC1), while potassium-chloride co-transporter 2 (KCC2) remained unchanged. Melatonin increased the expression of tryptophan hydroxylase 2 (TPH2, serotonin-synthesizing enzyme). The present study suggests that melatonin may promote its wakefulness by modulating NO-regulated WNK-SPAK/OSR1-KNCC1 signaling in rat DRN serotonergic neurons.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.3
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• pp.249-255
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• 2015

Wnk kinase maintains cell volume, regulating various transporters such as sodium-chloride cotransporter, potassium-chloride cotransporter, and sodium-potassium-chloride cotransporter 1 (NKCC1) through the phosphorylation of oxidative stress responsive kinase 1 (OSR1) and STE20/SPS1-related proline/alanine-rich kinase (SPAK). However, the activating mechanism of Wnk kinase in specific tissues and specific conditions is broadly unclear. In the present study, we used a human salivary gland (HSG) cell line as a model and showed that $Ca^{2+}$ may have a role in regulating Wnk kinase in the HSG cell line. Through this study, we found that the HSG cell line expressed molecules participating in the WNK-OSR1-NKCC pathway, such as Wnk1, Wnk4, OSR1, SPAK, and NKCC1. The HSG cell line showed an intracellular $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) increase in response to hypotonic stimulation, and the response was synchronized with the phosphorylation of OSR1. Interestingly, when we inhibited the hypotonically induced $[Ca^{2+}]_i$ increase with nonspecific $Ca^{2+}$ channel blockers such as 2-aminoethoxydiphenyl borate, gadolinium, and lanthanum, the phosphorylated OSR1 level was also diminished. Moreover, a cyclopiazonic acid-induced passive $[Ca^{2+}]_i$ elevation was evoked by the phosphorylation of OSR1, and the amount of phosphorylated OSR1 decreased when the cells were treated with BAPTA, a $Ca^{2+}$ chelator. Finally, through that process, NKCC1 activity also decreased to maintain the cell volume in the HSG cell line. These results indicate that $Ca^{2+}$ may regulate the WNK-OSR1 pathway and NKCC1 activity in the HSG cell line. This is the first demonstration that indicates upstream $Ca^{2+}$ regulation of the WNK-OSR1 pathway in intact cells.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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• 2009.11a
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• pp.76-76
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• 2009

PZT세라믹스는 높은 압전특성과 우수한 큐리온도($400^{\circ}C$)를 보유하고 있어 오래시간에 걸쳐 주목받고 있다. 현재 압전변압기, 액츄에이터, 센서등의 압전소자는 PZT를 이용하여 제작하고 있지만 PZT는 고온 소결시 PbO의 휘발이 환경오염을 초래하며 인체의 유해하다는 연구결과가 나왔다. 이에 최근에는 PbO가 포함 되지않은 무연(lead-free)계 압전세라믹스가 주목받고 있다. 무연 압전 세라믹스의 종류로는 Bi-layer-structured ceramics, Bi-perovskite type ceramics, NKN base ceramics 가 존재하고 있다. 그 중 $(Na_{0.5}K_{0.5})NbO_3(NKN)$ 세라믹스는 높은 큐리온도와($400^{\circ}C$)와 높은 전기기계 결합계수(약 36%)를 보유하고 있어 많은 연구가 이루어 지고 있다. 하지만 NKN은 PZT에 비하여 치밀성이 낮으며 일반적인 산화물 소결방법으로는 밀도를 높이기가 어려운 단점이 존재한다. 이를 개선하기 위한 방법으로 hot pressing와 spak plasma sintering, RTGG와 같은 방법으로 밀도를 높일수 있지만 비용이 많이 들어 일반적으로 사용이 어렵다. 다른 방법으로 NKN에 첨가물을 넣는 방법을 사용하고 있는데 방법으로 $LiNbO_3$, $LiTiO_3$, $LiSbO_5$를 첨가하여 개선하는 방법이 있다. 본 실험은 첨가물을 넣는 방식으로 비화학양론적 $(Na_{0.5}K_{0.5})_{0.97}(Nb_{0.9}Ta_{0.1})O_3(NKNT)$조성에 CuO를 mol%로 변화주어 유전 및 압전 특성을 조사하였다.

• The Korean Journal of Physiology and Pharmacology
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• v.22 no.1
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• pp.91-99
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• 2018

Protein phosphatase 1 (PP1) is involved in various signal transduction mechanisms as an extensive regulator. The PP1 catalytic subunit (PP1c) recognizes and binds to PP1-binding consensus residues (FxxR/KxR/K) in NBCe1-B. Consequently, we focused on identifying the function of the PP1-binding consensus residue, $^{922}FMDRLK^{927}$, in NBCe1-B. Using site-directed mutagenesis and co-immunoprecipitation assays, we revealed that in cases where the residues were substituted (F922A, R925A, and K927A) or deleted (deletion of amino acids 922-927), NBCe1-B mutants inhibited PP1 binding to NBCe1-B. Additionally, by recording the intracellular pH, we found that PP1-binding consensus residues in NBCe1-B were not only critical for NBCe1-B activity, but also relevant to its surface expression level. Therefore, we reported that NBCe1-B, as a substrate of PP1, contains these residues in the C-terminal region and that the direct interaction between NBCe1-B and PP1 is functionally critical in controlling the regulation of the ${HCO_3}^-$ transport. These results suggested that like IRBIT, PP1 was another novel regulator of ${HCO_3}^-$ secretion in several types of epithelia.