• 제목/요약/키워드: Calcium entry

검색결과 58건 처리시간 0.028초

Differential Vasorelaxant Effects of KR-30075, a New Cyclic AMP-phosphodiesterase Inhibitor, on Guinea-pig Pulmonary, Bovine Coronary and Renal Arteries

  • Jung, Yee-Suk;Kwon, Kwang-Il;Zee, Ok-Pyo
    • Archives of Pharmacal Research
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    • 제13권2호
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    • pp.136-141
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    • 1990
  • The vasorelaxant effects of KR-30075 in guinea-pig pulmonary, bovine coronary and renal arterial strips contracted by either$K^+$depolarization, phenylephrine, or prostaglandin $F_{2a}$($PGF_{2a}$) were evaluated. KR-30075 was more potent than imazodan as a vasorelaxant against $PGF_{2a}$-induced contractions in bovine coronary and renal arteries, whereas against$K^+$induced contractions KR-30075 was less potent than imazodan in guieapig pulmonary arteries and more potent in bovine coronary arteries. KR-30075 was more potent against contractions induced by phenylephrine or $PGF_{2a}$ than the contractions induced by $K^+$ This profile of activity for KR-30075 was similar to that of imazodan and dissimilar from the calcium entry blocking agent nifedipine. There was no vascular selectivity of KR-30075 between bovine coronary and renal arterial strip preparations. In conclusion, this study shows that KR-30075 represents the vasorelaxant effects on guinea-pig pulmonary, bovine coronary and renal arteries without specific vascular selectivity. The vasorelaxant profile of KR-30075, with different sources of vascular smooth muscle, is unlike that of calcium entry blocking agent and more similar to the profile of the agent that inhibit cyclic nucleotide phosphodiesterase.

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Regulation of the expression and function of TRPCs and Orai1 by Homer2 in mouse pancreatic acinar cells

  • Kang, Jung Yun;Kang, Namju;Yang, Yu-Mi
    • International Journal of Oral Biology
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    • 제46권3호
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    • pp.134-139
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    • 2021
  • Under physiological conditions, calcium (Ca2+) regulates essential functions of polarized secretory cells by the stimulation of specific Ca2+ signaling mechanisms, such as increases in intracellular Ca2+ concentration ([Ca2+]i) via the store-operated Ca2+ entry (SOCE) and the receptor-operated Ca2+ entry (ROCE). Homer proteins are scaffold proteins that interact with G protein-coupled receptors, inositol 1,4,5-triphosphate (IP3) receptors, Orai1-stromal interaction molecule 1, and transient receptor potential canonical (TRPC) channels. However, their role in the Ca2+ signaling in exocrine cells remains unknown. In this study, we investigated the role of Homer2 in the Ca2+ signaling and regulatory channels to mediate SOCE and ROCE in pancreatic acinar cells. Deletion of Homer2 (Homer2-/-) markedly increased the expression of TRPC3, TRPC6, and Orai1 in pancreatic acinar cells, whereas these expressions showed no difference in whole brains of wild-type and Homer2-/- mice. Furthermore, the response of Ca2+ entry by carbachol also showed significant changes to the patterns regulated by specific blockers of SOCE and ROCE in pancreatic acinar cells of Homer2-/- mice. Thus, these results suggest that Homer2 plays a critical role in the regulatory action of the [Ca2+]i via SOCE and ROCE in mouse pancreatic acinar cells.

Store-operated calcium entry in the satellite glial cells of rat sympathetic ganglia

  • Sohyun Kim;Seong Jun Kang;Huu Son Nguyen;Seong-Woo Jeong
    • The Korean Journal of Physiology and Pharmacology
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    • 제28권1호
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    • pp.93-103
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    • 2024
  • Satellite glial cells (SGCs), a major type of glial cell in the autonomic ganglia, closely envelop the cell body and even the synaptic regions of a single neuron with a very narrow gap. This structurally unique organization suggests that autonomic neurons and SGCs may communicate reciprocally. Glial Ca2+ signaling is critical for controlling neural activity. Here, for the first time we identified the machinery of store-operated Ca2+ entry (SOCE) which is critical for cellular Ca2+ homeostasis in rat sympathetic ganglia under normal and pathological states. Quantitative realtime PCR and immunostaining analyses showed that Orai1 and stromal interaction molecules 1 (STIM1) proteins are the primary components of SOCE machinery in the sympathetic ganglia. When the internal Ca2+ stores were depleted in the absence of extracellular Ca2+, the number of plasmalemmal Orai1 puncta was increased in neurons and SGCs, suggesting activation of the Ca2+ entry channels. Intracellular Ca2+ imaging revealed that SOCE was present in SGCs and neurons; however, the magnitude of SOCE was much larger in the SGCs than in the neurons. The SOCE was significantly suppressed by GSK7975A, a selective Orai1 blocker, and Pyr6, a SOCE blocker. Lipopolysaccharide (LPS) upregulated the glial fibrillary acidic protein and Toll-like receptor 4 in the sympathetic ganglia. Importantly, LPS attenuated SOCE via downregulating Orai1 and STIM1 expression. In conclusion, sympathetic SGCs functionally express the SOCE machinery, which is indispensable for intracellular Ca2+ signaling. The SOCE is highly susceptible to inflammation, which may affect sympathetic neuronal activity and thereby autonomic output.

Exposure to 835 MHz RF-EMF decreases the expression of calcium channels, inhibits apoptosis, but induces autophagy in the mouse hippocampus

  • Kim, Ju Hwan;Sohn, Uy Dong;Kim, Hyung-Gun;Kim, Hak Rim
    • The Korean Journal of Physiology and Pharmacology
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    • 제22권3호
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    • pp.277-289
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    • 2018
  • The exponential increase in the use of mobile communication has triggered public concerns about the potential adverse effects of radiofrequency electromagnetic fields (RF-EMF) emitted by mobile phones on the central nervous system (CNS). In this study, we explored the relationship between calcium channels and apoptosis or autophagy in the hippocampus of C57BL/6 mice after RF-EMF exposure with a specific absorption rate (SAR) of 4.0 W/kg for 4 weeks. Firstly, the expression level of voltage-gated calcium channels (VGCCs), a key regulator of the entry of calcium ions into the cell, was confirmed by immunoblots. We investigated and confirmed that pan-calcium channel expression in hippocampal neurons were significantly decreased after exposure to RF-EMF. With the observed accumulation of autolysosomes in hippocampal neurons via TEM, the expressions of autophagy-related genes and proteins (e.g., LC3B-II) had significantly increased. However, down-regulation of the apoptotic pathway may contribute to the decrease in calcium channel expression, and thus lower levels of calcium in hippocampal neurons. These results suggested that exposure of RF-EMF could alter intracellular calcium homeostasis by decreasing calcium channel expression in the hippocampus; presumably by activating the autophagy pathway, while inhibiting apoptotic regulation as an adaptation process for 835 MHz RF-EMF exposure.

MDCK세포의 tight junction 형성이 Toxoplusmu gondii의 숙주세포 침투에 미치는 효과 (Tight junctional inhibition of entry of Toxoplasma gondii into MDCK cells)

  • 남호우;윤지혜
    • Parasites, Hosts and Diseases
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    • 제28권4호
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    • pp.197-206
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    • 1990
  • MDCK세포간에 형성된 tight junction이 Toxoplasma gondii이 숙주세포 침투에 미치는 영향에 대하여 고찰 하기 위해 여러 조건의 세포배양을 시행하였다. MDCK세포를 25-well배양기 내의 18-mm cover glass에 $1{\times}10^5,{\;}3{\times}10^5{\;}및{\;}5{\times}10^5$ 개로 분주한 후 배양 1, 2, 3 및 4일에 다수의 Toxoplasma를 첨가하여 1시간 동안 배양한 다음 Giemsa 용액으로 염색한 후 관찰하였을 때, 각 실험군에서 침투한 원충의 숫자는 숙주세포가 포화밀도를 이루면서 급격히 감소하였다. 배양액 내의 calcium 농도를 일반적인 1.8mM에서 $5{\;}{\mu}M$로 낮추어 포화밀도의 MDCK 세포간에 tight junction 형성을 억제하였을 때, 원충의 침투가 약 2배 증가하였으며(p<0.05) 포화밀도 이전의 배양에서는 원충의 침투가 감소하였다. Trypsin-EDTA를 처리하여 포화밀도의 배양에서 tight junction을 소화 시킨 경우, 원충의 침투가 약 2.5배 증가하였으며 (p<0.05) 포화밀도 이전의 배양에서는 급격히 감소하였다. 이상의 결과들로 볼 때, 포화밀도의 MDCK세포간에 형성된 tight junction이 Toxoplasma의 침투를 억제하는 것을 알 수 있었다. 이는 Toxoplasma가 숙주세포로 침투할 때 숙주세포의 막구조에 대한 특이성이 있음을 시사 하며, 상피세포에서는 tight junction 위의 막(apical membrane) 보다 옆 및 아래의 막(basolateral membrane)상의 구조를 이용하는 것으로 추정되었다.

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Presenilin Modulates Calcium-permeant, Magnesium-Nucleotide regulated channel, I(MgNUM)

  • Shin, Sun-Young;Jeong, Soon-Youn;Uhm, Dae-Yong;Sungkwon Chung
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2003년도 정기총회 및 학술발표회
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    • pp.47-47
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    • 2003
  • The presenilin 1 (PS1) or PS2 is an essential component of the ${\gamma}$-secretase complex, which mediates the intramembrane proteolysis of selected type-I membrane, including the ${\beta}$-amyloid precursor protein (APP) to yield A${\beta}$. Familial Alzheimer's disease (FAD)-associated mutations in presenilins give rise to an increased production of a highly amyloidogenic A${\beta}$42. In addition to their well-documented proteolytic function, the presenilins play a role in calcium signaling. We have previously reported that presenilin FAD mutations cause highly consistent alterations in intracellular calcium signaling pathways, which include deficits in capacitative calcium entry (CCE), the refilling mechanism for depleted internal calcium stores. However, molecular basis for the presenilin-mediated modulation of CCE remains to be elucidated. In the present study, whole-cell patch clamp method was used to identify a specific calcium-permeable ion channel current(s) that is responsible for the CCE deficits associated with FAD-linked PS1 mutants. Unexpectedly, both voltage-activated and conventional store depletion-activated calcium currents I(CRAC), were absent in HEK293 cells, which were stably transfected either with wild-type or FAD mutant (L286V, M146L, and delta E9) forms of PS1. Recently, magnesium-nucleotide-regulated metal cation current, or I(MagNum), has been described and appears to share many common properties with I(CRAC) including calcium permeability and inhibitor sensitivity (e.g. 2-APB). We have detected I(MagNum) in all 293 cells tested. Interestingly, FAD mutant 293 cells developed only about half of currents compared to PS1 wild type cells.

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Caffeine and 2-Aminoethoxydiphenyl Borate (2-APB) Have Different Ability to Inhibit Intracellular Calcium Mobilization in Pancreatic Acinar Cell

  • Choi, Kyung-Jin;Kim, Kab-Sung;Kim, Se-Hoon;Kim, Dong-Kwan;Park, Hyung-Seo
    • The Korean Journal of Physiology and Pharmacology
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    • 제14권2호
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    • pp.105-111
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    • 2010
  • Inositol 1,4,5-trisphosphate receptors ($InsP_3Rs$) modulate $Ca^{2+}$ release from intracellular $Ca^{2+}$ store and are extensively expressed in the membrane of endoplasmic/sarcoplasmic reticulum and Golgi. Although caffeine and 2-aminoethoxydiphenyl borate (2-APB) have been widely used to block $InsP_3Rs$, the use of these is limited due to their multiple actions. In the present study, we examined and compared the ability of caffeine and 2-APB as a blocker of $Ca^{2+}$ release from intracellular $Ca^{2+}$ stores and $Ca^{2+}$ entry through store-operated $Ca^{2+}$ (SOC) channel in the mouse pancreatic acinar cell. Caffeine did not block the $Ca^{2+}$ entry, but significantly inhibited carbamylcholine (CCh)-induced $Ca^{2+}$ release. In contrast, 2-APB did not block CCh-induced $Ca^{2+}$ release, but remarkably blocked SOC-mediated $Ca^{2+}$ entry at lower concentrations. In permeabilized acinar cell, caffeine had an inhibitory effect on InsP3-induced $Ca^{2+}$ release, but 2-APB at lower concentration, which effectively blocked $Ca^{2+}$ entry, had no inhibitory action. At higher concentrations, 2-APB has multiple paradoxical effects including inhibition of Ins$P_3$-induced $Ca^{2+}$ release and direct stimulation of $Ca^{2+}$ release. Based on the results, we concluded that caffeine is useful as an inhibitor of $InsP_3R$, and 2-APB at lower concentration is considered a blocker of $Ca^{2+}$ entry through SOC channels in the pancreatic acinar cell.

Regulatory mechanisms of the store-operated Ca2+ entry through Orai1 and STIM1 by an adaptor protein in non-excitable cells

  • Kang, Jung Yun;Yang, Yu-Mi
    • International Journal of Oral Biology
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    • 제47권3호
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    • pp.33-40
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    • 2022
  • Store-operated Ca2+ entry (SOCE) represents one of the major Ca2+ entry routes in non-excitable cells. It is involved in a variety of fundamental biological processes and the maintenance of Ca2+ homeostasis. The Ca2+ release-activated Ca2+ (CRAC) channel consists of stromal interaction molecule and Orai; however, the role and action of Homer proteins as an adaptor protein to SOCE-mediated Ca2+ signaling through the activation of CRAC channels in non-excitable cells still remain unknown. In the present study, we investigated the role of Homer2 in the process of Ca2+ signaling induced by the interaction between CRACs and Homer2 proteins in non-excitable cells. The response to Ca2+ entry by thapsigargin-mediated Ca2+ store depletion remarkably decreased in pancreatic acinar cells of Homer2-/- mice, as compared to wild-type cells. It also showed critical differences in regulated patterns by the specific blockers of SOCE in pancreatic acinar cells of Homer2-/- mice. The response to Ca2+ entry by the depletion in Ca2+ store markedly increased in the cellular overexpression of Orai1 and STIM1 as compared to the overexpression of Homer2 in cells; however, this response was remarkably inhibited by the overexpression of Orai1, STIM1, and Homer2. These results suggest that Homer2 has a critical role in the regulatory action of SOCE activity and the interactions between CRAC channels.

SIMULTANEOUS EXPRESSION OF HUMAN CYTOCHROME P450 3A5 AND NADPH-CYTOCHROME P450 REDUCTASE IN CHINESE HAMSTER OVARY CELL USING INTERNAL RIBOSOME ENTRY SITE

  • Kang, Hyuck-Joon;Kang, Jin-Sun;Dong, Mi-Sook;Park, Chang-Hwan
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
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    • pp.189-189
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    • 2001
  • For a continuous expression of human cytochrome p450 3A5 (CYP3A5) and NADPH-cytochrome P450 reductase (CYPR) proteins, bicistronic construct (CYP3A5BC-LNCX2) was made using internal ribosome entry site (IRES). As for mammalian cell expression, we used pLNCX2 retroviral vector; and using calcium phosphate, plasmid transfer was achieved in 293GPG cell and transduced in Chinese hamster ovary (CHO) cell.(omitted)

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