• Journal of Microbiology and Biotechnology
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• 제1권3호
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• pp.151-156
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• 1991

In order to construct a yeast strain having high ethanol tolerance together with good lactose fermentation ability, the protoplast fusion using Saccharomyces cerevisiae STV 89 and Kluyveromyces fragilis CBS 397 was carried out. Auxotrophic mutants of K. fragilis were obtained as a selection marker by treatment of ethylmethane sulfonate. The best mutant for protoplast fusion was selected based on the capabilities of ${\beta}-galactosidase$ production and lactose fermentation. The protoplast fusion using polyethylene glycol and calcium chloride solution led to the fusion frequence of $3{\times}10^{-6}$ and a number of fusants were obtained. Among these fusants, a fusant F-3-19 showed the best results in terms of ethanol tolerance, ${\beta}-galactosidase$ activity and lactose fermentation. The performance of lactose fermentation and ethanol tolerance by this fusant were better than those of K. fragilis. Study on the ethanol tolerance having relation to fatty acid composition and intracellular ethanol concentration revealed that the fusant F-3-19 had a higher unsaturated fatty acids content and accumulated less amount of intracellular ethanol compared with a parent of K. fragilis.

• The Korean Journal of Physiology and Pharmacology
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• 제3권4호
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• pp.439-445
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• 1999

This study is to investigate the mechanism of inhibitory effect of imipramine on the calcium utilization in single cells isolated from canine detrusor. 2 mm thick smooth muscle chops were incubated in 0.12% collagenase solution at $36^{circ}C,$ and aerated with 95% $O_2/5%\;CO_2,$ and then cell suspension was examined. Acetylcholine (ACh) evoked a concentration-dependent contraction of the isolated detrusor cells in normal physiologic salt solution (PSS), and the ACh-induced contraction was significantly inhibited by imipramine. In $Ca^{2+}-free$ PSS, ACh-induced contraction was less than those in normal PSS and it was not affected by the pretreatment with imipramine. $Ca^{2+}-induced$ contraction in $Ca^{2+}-free$ PSS was supressed by imipramine, but addition of A 23187, a calcium ionophore, overcomed the inhibitory effect of imipramine. High potassium-depolarization (40 mM KCl) evoked cell contraction, which was inhibited by imipramine. Caffeine, a releasing agent of the stored $Ca^{2+}$ from sarcoplasmic reticulum, evoked a contraction of the cells that was not blocked by the pretreatment with imipramine. These results suggest that imipramine inhibits the influx of calcium in the detrusor cells through both the receptor-operated- and voltage-gated-calcium channels, but does not affect the release of calcium from intracellular storage site.

• The Korean Journal of Physiology and Pharmacology
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• 제12권3호
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• pp.95-99
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• 2008

Calcium ($Ca^{2+}$) is an intracellular second messenger associated with neuronal plasticity of the central nervous system. The calcium-binding proteins regulate the $Ca^{2+}$-mediated signals in the cytoplasm and buffer the calcium concentration. This study examined temporal changes of three calcium-binding proteins (calretinin, calbindin and parvalbumin) in the medial vestibular nucleus (MVN) during vestibular compensation after unilateral labyrinthectomy (UL) in rats. Rats underwent UL, and the changes in the expression of these proteins at 2, 6, 12, 24, 48, and 72 h were examined by immuno-fluorescence staining. The expression levels of all three proteins increased immediately after UL and returned to the control level by 48 h. However, the level of calretinin showed changes different from the other two proteins, being expressed at significantly higher level in the contralateral MVN than in the ipsilateral MVN 2 h after UL, whereas the other two proteins showed similar expression levels in both the ipsilateral and contralateral MVN. These results suggest that the calcium binding proteins have some protective activity against the increased $Ca^{2+}$ levels in the MVN. In particular, calretinin might be more responsive to neuronal activity than calbindin or parvalbumin.

• The Korean Journal of Physiology and Pharmacology
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• 제20권4호
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• pp.387-397
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• 2016

Neurofibrillary tangles (NFTs) of microtubule-associated protein tau are a pathological hallmark of Alzheimer's disease (AD). Endoplasmic reticulum (ER) stress has been known to be involved in the pathogenesis of AD. However, the exact role of ER stress in tau pathology has not yet been clearly elucidated. In present study, the possible relationship between tau pathology and ER stress was examined in terms of sorcin, which is a calcium binding protein and plays an important role in calcium homeostasis. Our previous yeast two hybrid study showed that sorcin is a novel tau interacting protein. Caspase-3-cleaved tau (T4C3) showed significantly increased tau-sorcin interaction compared to wild type tau (T4). Thapsigargin-induced ER stress and co-expression of constitutively active $GSK3{\beta}$ ($GSK3{\beta}-S9A$) also exhibited significantly increased tau-sorcin interactions. T4C3-expressing cells showed potentiated thapsigargin -induced apoptosis and disruption of intracellular calcium homeostasis compared to T4-expressing cells. Overexpression of sorcin significantly attenuated thapsigargin-induced apoptosis and disruption of calcium homeostasis. In contrary, siRNA-mediated knock-down of sorcin showed significantly increased thapsigargin-induced apoptosis and disruption of calcium homeostasis. These data strongly suggest that sequestration of sorcin by aberrant forms of tau compromises the function of sorcin, such as calcium homeostasis and cellular resistance by ER stress, which may consequently result in the contribution to the progression of AD.

• Archives of Pharmacal Research
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• 제18권5호
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• pp.295-300
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• 1995

These studies were designed to examine the effects of ginsenosides on glutamate neurotansmission. In primary cultures of rat cerebellar granule cells, ginsenosides (Rb1, Rc did not Rg1, $500\mug/ml$) increased glutamate release which was measured by HPLC. but HPLC, but Re did not shwo an elevation of glutamate release. However, all of these ginsenosides down-regulated N-methyl-D-aspartate (NMDA)-induced glutamate release. Rc strongly increased glutamate release and elevated intracellular clcium concentrations $([Ca_{2+}]_i)$ which was measured by ratio fluorometry with FURA-2AM. These results indicate that ginsenosides have a homeostatic effect on glutamate neurotransmission, and there is a structure-function relationship among the ginsenosides tested.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 2001년도 학술 발표회 진행표 및 논문초록
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• pp.49-49
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• 2001

Sphingosine 1-phosphate is a metabolite of complex sphingolipids that acts as both a second messenger and as a high-affinity ligand for cell surface receptor. Since the possible involvement of sphingosine 1-phosphate has not been investigated in adipocyte, we examined the response of intracellular calcium ([Ca$^{2＋}$]$_{i}$) and intracellular cAMP ([cAMP]$_{i}$ and the effect of sphingosine 1-phosphate on adipocyte function using rat primary adipocyte.(omitted)ted)

• 대한약리학회지
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• 제28권1호
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• pp.81-89
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• 1992

항우울제에 속하는 imipramine은 유뇨증에 대한 치료제로서도 널리 사용되고 있다. Imipramine이 유뇨증 치료제로서의 명백한 치료효과를 보이기는 하나, 그 작용기전에 대해서는 아직 논란이 많다. 그 중 가장 유력한 설은 말초성 자율신경에 대한 작용으로서 무스카린성 길항작용과 교감신경말단에서의 catecholamine 재섭취 방해작용 및 직접적인 방광근 이완 작용 등이 제시되고 있다. 또 최근에는 방광 평활근에 대한 직접작용에 칼슘 이동 억압 작용이 중요한 역할을 한다고 주장되고 있다. 이에 본 실험에서는 흰쥐의 적출방광 배뇨근 절편을 사용하여 imipramine의 항유뇨작용과 calcium동원과의 관련성을 추구하여 다음과 같은 결과를 얻었다. 1. 배뇨근 절편은 전기장자극에 의해 주파수의존적으로 수축하였는데, 이 전기장자극 유발 수축반응은 imipramine에 의해 농도 의존적으로 억제되었고 고농도에서는 완전히 소실되었으나, atropine에 의해서는 고농도에서도 소실되지 않았다. 2. Imipramine은 자발수축 및 기본 장력에 대하여 이들을 농도 의존적인 양상으로 억제하였으며, 이 억제작용은 diltiazem에 의한 억제와 유사하였다. 그러나 atropine은 배뇨근절편의 기본장력이나 자발수축에 아무런 영향도 미치지 않았다. 3. $^*Imipramine$은 bethanechol 유발수축과 adenosine triphosphate (ATP) 유발수축을 농도의존적으로 억압하였다. 4. Imipramine은 칼슘배제용액에서 칼슘 첨가에의한 수축성의 회복을 억제하였는데, 이러한 작용은 diltiazem보다 그 효력이 낮았으나 그 작용양상은 유사하였다. 5. Imipramine에 의해 감소되었던 배뇨근절편의 기본장력은 calcium ionophore인 A23187에 의해 회복되었다. 칼슘배제 영양액에 장시간 노출됨으로써 기본장력은 소실되었으며, imipramine을 전처치한 경우는 A23187첨가에 의해 장력이 회복되지 않았으나, 세포내칼슘 유리억제제인 trimethoxybenzoic acid 8-(diethylamino)octyl ester [TMB-8]을 전처치한 경우 A23187은 배뇨근의 기본장력을 현저히 회복시켰다. 이상의 결과로 미루어 보아 imipramine의 배뇨근 수축 억제작용의 기전에는 무스카린성 및 퓨린성 수용체봉쇄작용도 관여하나, 주된 기전은 평활근세포에 직접 작용하여 세포외 칼슘의 유입을 억제하는 것으로 사료된다.

• The Korean Journal of Physiology and Pharmacology
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• 제12권2호
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• pp.43-49
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• 2008

Flavonoids have been shown to affect calcium signaling in neurons. However, there are no reports on the effect of apigenin on glutamate-induced calcium signaling in neurons. We investigated whether apigenin affects glutamate-induced increase of free intracellular $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) in cultured rat hippocampal neurons, using fura-2-based digital calcium imaging and microfluorimetry. The hippocampal neurons were used between 10 and 13 days in culture from embryonic day 18 rats. Pretreatment of the cells with apigenin ($1{\mu}M$ to $100{\mu}M$) for 5 min inhibited glutamate ($100{\mu}M$, 1 min) induced $[Ca^{2+}]_i$ increase, concentration-dependently. Pretreatment with apigenin ($30{\mu}M$) for 5 min significantly decreased the $[Ca^{2+}]_i$ responses induced by two ionotropic glutamate receptor agonists, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic (AMPA, $10{\mu}M$, 1 min) and N-methyl-D-aspartate (NMDA, $100{\mu}M$, 1 min), and significantly inhibited the AMPA-induced peak currents. Treatment with apigenin also significantly inhibited the $[Ca^{2+}]_i$ response induced by 50 mM KCl solution, decreased the $[Ca^{2+}]_i$ responses induced by the metabotropic glutamate receptor agonist, (S)-3,5-dihydroxy-phenylglycine (DHPG, 100 $[Ca^{2+}]_i$, 90 s), and inhibited the caffeine (10 mM, 2 min)-induced $[Ca^{2+}]_i$ responses. Furthermore, treatment with apigenin ($30{\mu}M$) significantly inhibited the amplitude and frequency of 0.1 mM $[Mg^{2+}]_o$-induced $[Ca^{2+}]_i$ spikes. These data together suggest that apigenin inhibits glutamate-induced calcium signaling in cultured rat hippocampal neurons.

• Molecules and Cells
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• 제25권3호
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• pp.390-396
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• 2008

Calreticulin (CRT) is one of the major $Ca^{2+}$ binding chaperone proteins of the endoplasmic reticulum (ER) and an unusual luminal ER protein. Postnatally elevated expression of CRT leads to impaired development of the cardiac conductive system and may be responsible for the pathology of complete heart block. In this study, the molecular mechanisms that affect $Ca^{2+}$-dependent signal cascades were investigated using CRT-overexpressing cardiomyocytes. In particular, we asked whether calreticulin plays a critical role in the activation of $Ca^{2+}$-dependent apoptosis. In the cells overexpressing CRT, the intracellular calcium concentration was significantly increased and the activity of PKC and level of SECAR2a mRNA were reduced. Phosphorylation of Akt and ERKs decreased compared to control. In addition the activity of the anti-apoptotic factor, Bcl-2, was decreased and the activities of pro-apoptotic factor, Bax, p53 and caspase 8 were increased, leading to a dramatic augmentation of caspase 3 activity. Our results suggest that enhanced CRT expression in mature cardiomyocytes disrupts intracellular calcium regulation, leading to calcium-dependent apoptosis.

• 대한본초학회지
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• 제29권4호
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• pp.83-89
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• 2014

Objectives : The purpose of this study was to investigate effects of Houttuyniae Herba water extract (HC) on calcium release and production of various inflammatory mediators such as nitric oxide (NO), interferon-inducible protein (IP)-10, platelet derived growth factor (PDGF)-BB, keratinocyte-derived chemokine (KC), vascular endothelial growth factor (VEGF), interleukin (IL)-4, and IL-5 in lipopolysaccharide (LPS)-induced RAW 264.7 mouse macrophages. Methods : NO production was measured by Griess reagent assay. Intracellular calcium level was measured with Fluo-4 assay. Levels of cytokines were measured by High-throughput multiplex bead array cytokine assay based on xMAP (multi-analyte profiling beads) technology. Results : HC significantly decreased NO production for 24 hrs incubation at the concentrations of 10, 25, 50, 100, and $200{\mu}/mL$ in LPS-induced RAW 264.7 (P < 0.05). HC significantly decreased production of IP-10, KC, VEGF, and PDGF-BB for 24 hrs incubation at the concentrations of 50, 100, and $200{\mu}/mL$ in LPS-induced RAW 264.7 (P < 0.05). HC also significantly decreased intracellular calcium release for 24 hrs incubation at the concentrations of 25, 50, 100, and $200{\mu}/mL$ in LPS-induced RAW 264.7 (P < 0.05). But HC did not show any significant effect on production of IL-4 and IL-5 in LPS-induced RAW 264.7. Conclusions : The results suggested that HC has anti-inflammatory property related with its inhibition on the production of NO, IP-10, KC, VEGF, and PDGF-BB in LPS-induced macrophages via calcium pathway.