• The Korean Journal of Physiology and Pharmacology
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• 제20권1호
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• pp.75-82
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• 2016

Paroxetine, a selective serotonin reuptake inhibitor (SSRI), has been reported to have an effect on several ion channels including human ether-a-go-go-related gene in a SSRI-independent manner. These results suggest that paroxetine may cause side effects on cardiac system. In this study, we investigated the effect of paroxetine on Kv1.5, which is one of cardiac ion channels. The action of paroxetine on the cloned neuronal rat Kv1.5 channels stably expressed in Chinese hamster ovary cells was investigated using the whole-cell patch-clamp technique. Paroxetine reduced Kv1.5 whole-cell currents in a reversible concentration-dependent manner, with an $IC_{50}$ value and a Hill coefficient of $4.11{\mu}M$ and 0.98, respectively. Paroxetine accelerated the decay rate of inactivation of Kv1.5 currents without modifying the kinetics of current activation. The inhibition increased steeply between -30 and 0 mV, which corresponded with the voltage range for channel opening. In the voltage range positive to 0 mV, inhibition displayed a weak voltage dependence, consistent with an electrical distance ${\delta}$ of 0.32. The binding ($k_{+1}$) and unbinding ($k_{-1}$) rate constants for paroxetine-induced block of Kv1.5 were $4.9{\mu}M^{-1}s^{-1}$ and $16.1s^{-1}$, respectively. The theoretical $K_D$ value derived by $k_{-1}/k_{+1}$ yielded $3.3{\mu}M$. Paroxetine slowed the deactivation time course, resulting in a tail crossover phenomenon when the tail currents, recorded in the presence and absence of paroxetine, were superimposed. Inhibition of Kv1.5 by paroxetine was use-dependent. The present results suggest that paroxetine acts on Kv1.5 currents as an open-channel blocker.

• Reproductive and Developmental Biology
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• 제33권4호
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• pp.203-209
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• 2009

The present study was performed to identify changes of plasminogen activator (PA) and plasminogen activator inhibitor (PAI) in porcine oviduct epithelial cells (POECs) during the estrous cycle. POECs obtained from ovary in pre-ovulatory (Pre-Ov), early to mid-luteal stage (Early-mid L) and post-ovulatory stage (Post-Ov). For the examine of PA activity, $1{\times}10^5$ fresh cells of POECs were cultured in DMEM/Ham F-12 containing 10% FBS and 0.2% amphotericin under humidified atmosphere of 5% $CO_2$ in air and $38^{\circ}C$. The urokinase-type PA (uPA) was observed at 7 days of POECs culture. PA activity was measured with culture prolonged of 0, 3, 6, 12 and 24 h after culture of 7 days. The PA activity were high significantly (p<0.05) at 12 h of culture, but PA activity were decreased with culture periods increased. The PA activity in POECs of Post-Ov stage were higher significantly (p<0.05) than that of Early-mid L and Pre-Ov stage. When PAI-1 and PAI-2 were added during the POECs culture, the PA were observed significant low activity (p<0.05). The PA activity and protein expression were decreased by PA inhibitor. This results suggest that PAI-1 and PAI-2 have a suppressive action on change of PA activity during the estrous cycle of pigs. Specifically, this study using PA inhibitor was effect the PA activity and PAI expression in oviduct epithelial cells in pigs.

• Reproductive and Developmental Biology
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• 제34권4호
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• pp.289-297
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• 2010

Erythropoietin (EPO) is a glycoprotein hormone secreted from primarily cells of the peritubular capillary endothelium of the kidney, and is responsible for the regulation of red blood cell production. We constructed and expressed dimeric cDNAs in Chinease hamster ovary (CHO) cells encoding a fusion protein consisting of 2 complete human EPO domains linked by a 2-amino acid linker (Ile-Asp). We described the activity of dimeric hyperglycosylated EPO (dHGEPO) mutants containing additional oligosaccharide chains and characterized the function of glycosylation. No dimeric proteins with mutation at the $105^{th}$ amino acid were found in the cell medium. Growth and differentiation of the human EPO-dependent leukemiae cell line (F36E) were used to measure cytokine dependency and in vitro bioactivity of dHGEPO proteins. MIT assay at 24 h increased due to the survival of F36E cells. The dHGEPO protein migrated as a broad band with an average molecular mass of 75 kDa. The mutant, dHGEPO, was slightly higher than the wild-type (WT) dimeri-EPO band. Enzymatic N-deglycosylation resulted in the formation of a narrow band with a molecular mass twice of that of of monomeric EPO digested with an N-glycosylation enzyme. Hematocrit values were remarkably increased in all treatment groups. Pharmacokinetic analysis was also affected when 2.5 IU of dHGEPO were intravenously injected into the tails of the mice. The biological activity and half-life of dHGEPO mutants were enhanced as compared to the corresponding items associated the WT dimeric EPO. These results suggest that recombinant dHGEPO may be attractive biological and therapeutic targets.

• Journal of Life Science
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• 제11권2호
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• pp.103-107
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• 2001

The equine chorionic gonadotropin (eCG) subunits $\alpha$ and ${\beta}$ are transcribed from different genes and associate noncovalently to form the bioactive eCG heterodimer. Dimerization is rate limiting for eCG secretion, and dissociation leads to hormone inactivation. The correct conformation of the heterodimer is alto important for efficient secretion, hormone-specific post-translational modifications, receptor binding and signal transduction. To determine whether ${\alpha}$ and ${\beta}$ subunits can be synthesized as a single polypeptide chain (tethered-eCG) and also display biological activity, the tethered-eCG molecule by fusing the carboxyl terminus of the eCG ${\beta}$-subunit to the amino terminus of the af-subunit was construe-ted and transfected into chinese hamster ovary (CHO-Kl) cells. LH- and FSH-like activities were assayed in terms of testosterone production and aromatase activity in primary cultured rat Leydig cells and granulosa cells, respectively. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG ${\alpha}$/${\beta}$ and native eCG. FSH-like activity of the tethered-eCG was also shown similarly in comparison with the native and wild type eCG ${\alpha}$/${\beta}$. Our data for the first time suggest that the tethered-eCG can be expressed efficiently and the produced product by the CHO-K1 cells is fully LH- and FSH-like activities in rat in vitro bioassay system. Our results also suggest that this molecular can imply particular models of FSH-like activity not LH-like activity in the eCG. Taken together, these data indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion.

• 한국자원식물학회지
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• 제3권1호
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• pp.31-40
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• 1990

Pericarp and seedcoat removal treatments were tested to determine their effectiveness in the hard-to-germinate botanical seeds of jerusalem artichoke. Fresh seed of five Helianthus tuberosus L.varieties were (A)untreated (B)water soaked overnight (C) soaked overnight followed by renoval of pericarp or (D) soaked overnight followed by renoval of pericarp and seedcoat. The results indicate that treatments which removed the pericarp and seedcoat were the very effective, giving germination of over 90%. A considerable increase in germination did not follow only the peri-carp removal treatments. The factors inhibitory to germination of fresh jerusalem artichoke seed is associated withthe seedcoat. The removal treatment of pericarp and seed-cost is recommended despite its complexity because it givesa high germination percentage and varies least from varietyto variety. Heliarthus tuberosus L., a biomass potential crop, is a member of the family compositae. The genus Helianthushas provided man with two food plants, the sunflower (H.annus) and the jerusalem artichoke or topinambour (H. tube-rosus) . (3) The jerusalem artichoke grown for its tubers, has always been an extremely minor crop, but it is stillgrown in many places as a food for man or livestock and forthe production of alcohoL. Though tubers are used for pro-pagation jerusalem artichoke also flower and produce seedin head-like in florescences. Flowers are developed acro-petaLly on flattened receptacles such that outermost flowers are oldest. Each of these epigynous flowers may develop an achene-type fruit in which outer layers of the overy wallpersist, while inner layers become disorganized. Insidethe ovary wall of mature fruit, there is a papery seedcoat, probably composed of compact cells from endosperm, integu-ments, and nucellus.In general, the efforts to improving this crop havebeen hampered by the hard-to-germinate botanical seed.Seeds did not germinate for at least IL months after harvest.Fresh seeds of some varieties require one year more to gar-minate. (5) Since the time factor between generations isof concern in a prospective breeding program of jerusalemartichoke , these observations led to investigation of thenature of delayed seed germination in jerusalem artichokeas a biomass potential crop.

• Clinical and Experimental Reproductive Medicine
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• 제26권1호
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• pp.67-73
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• 1999

The insulin-like growth factor (IGF)s are believed to one of several growth factors that play an adjunctive role in ovarian follicular development. These factors circulate bound to a family of IGF-binding protein (IGFBP)s. It is known that circulating IGFBPs are involved in the transport of IGFs to tissues and modulate IGFs actions at local tissue. The purposes of this study were to evaluate changes in serum IGFBPs profiles during normal ovulatory menstrual cylce and to compare serum IGFBPs profiles in periovulatory phase of between normal ovulatory menstrual cylce and controlled hyperstimulated cycle. Fasting blood samples were obtained from 15 normal healthy women throughout normal ovulatory menstural cyle and on the day of aspiration of oocyte from 10 patients undergoing ovarian hyperstimuation for in vito fertilization-embryo transfer. Serum IGFBP-1 - IGFBP-4 were measured by western ligand blot and immunoprecipitation. Serum $17{\beta}$-estradiol was determined by radioimmunoassay. Type and molecular weight of serum IGFBP did not changed during normal ovulatory menstural cycle. No significant variation in the relative proportion and level of each IGFBP was found throughout normal ovulatory menstural cyle. Also, the relative proportion and level of each IGFBP did not correlated with serum $17{\beta}$-estradiol level. There was no significant difference in the relative proportion and level of each serum IGFBP between on the day of ovulation in normal ovulatory menstrual cylce and on the day of aspiration of oocyte in controlled hyperstimulated cycle. Our data indicate that IGFBPs have regulatory functions in ovary through an paracrine and autocrine rather than endocrine mechanism during normal ovulatory menstural cycle.

• The Korean Journal of Physiology and Pharmacology
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• 제10권2호
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• pp.71-77
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• 2006

The goal of this study was to analyze the effects of genistein, a widely used tyrosine kinase inhibitor, on cloned Shaw-type $K^+$ currents, Kv3.1 which were stably expressed in Chinese hamster ovary (CHO) cells, using the whole-cell configuration of patch-clamp techniques. In whole-cell recordings, genistein at external concentrations from 10 to $100{\mu}M$ accelerated the rate of inactivation of Kv3.1 currents, thereby concentration-dependently reducing the current at the end of depolarizing pulse with an $IC_{50}$ value of $15.71{\pm}0.67{\mu}M$ and a Hill coefficient of $3.28{\pm}0.35$ (n=5). The time constant of activation at a 300 ms depolarizing test pulses from -80 mV to +40 mV was $1.01{\pm}0.04$ ms and $0.90{\pm}0.05$ ms (n=9) under control conditions and in the presence of $20{\mu}M$ genistein, respectively, indicating that the activation kinetics was not significantly modified by genistein. Genistein $(20{\mu}M)$ slowed the deactivation of the tail current elicited upon repolarization to -40 mV, thus inducing a crossover phenomenon. These results suggest that drug unbinding is required before Kv3.1 channels can close. Genistein-induced block was voltage-dependent, increasing in the voltage range $(-20\'mV{\sim}0\'mV)$ for channel opening, suggesting an open channel interaction. Genistein $(20{\mu}M)$ produced use-dependent block of Kv3.1 at a stimulation frequency of 1 Hz. The voltage dependence of steady-state inactivation of Kv3.1 was not changed by $20{\mu}M$ genistein. Our results indicate that genistein blocks directly Kv3.1 currents in concentration-, voltage-, time-dependent manners and the action of genistein on Kv3.1 is independent of tyrosine kinase inhibition.

• 한국자원식물학회지
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• 제28권2호
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• pp.235-242
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• 2015

국내 도입된 하이부쉬 블루베리의 품종 선발의 기초 자료로 활용하기 위하여 블루베리의 형태적 및 품종별 화기 특성에 관해 연구하였다. 블루베리의 수술의 형태를 보면 화사는 판형이었고 모용이 잘 발달되어 있으며, 개약 시 약벽이 열개되지 않고 돌출된 두 개의 관으로 꽃가루가 배출되어 독특하였다. 화분은 3월 하순경에 감수 분열된 후에 분리되지 않은 채로 성숙한 사분자 화분(tetrad pollen)이었다. 약당 사분자 화분립 수는 400~1,300개였으며, 화분립 수가 많은 품종이 발아율도 높은 편이었다. 블루베리의 암술은 5개의 심피로 구성되어 있으며, 화주는 갈라진 부분이 없이 하나였다. 자방당 배주 수는 39~67개 정도였으며, 각 품종별로 변이 계수가 11.6~31.0%로 큰 편이었다. ‘Bluejay’와 ‘Sharpblue’간의 자가 및 타가교배 결과 자가교배보다 타가교배에서 종자 형성률이 높았다.

• 한국발생생물학회지:발생과생식
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• 제25권3호
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• pp.133-143
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• 2021

In contrast to the human lutropin receptor (hLHR) and rat LHR (rLHR), very few naturally occurring mutants in other mammalian species have been identified. The present study aimed to delineate the mechanism of signal transduction by three constitutively activating mutants (designated M410T, L469R, and D590Y) and two inactivating mutants (D383N and Y546F) of the eel LHR, known to be naturally occurring in human LHR transmembrane domains. The mutants were constructed and measured cyclic adenosine monophosphate (cAMP) accumulation via homogeneous time-resolved fluorescence assays in Chinese hamster ovary (CHO)-K1 cells. The activating mutant cells expressing eel LHR-M410T, L469R, and D590Y exhibited a 4.0-, 19.1-, and 7.8-fold increase in basal cAMP response without agonist treatment, respectively. However, inactivating mutant cells expressing D417N and Y558F did not completely impaired signal transduction. Specifically, signal transduction in the cells expressing activating mutant L469R was not occurred with a further ligand stimulation, showing that the maximal response exhibited approximately 53% of those of wild type receptor. Our results suggested that the constitutively activating mutants of the eel LHR consistently occurred without agonist treatment. These results provide important information of LHR function in fish and regulation with regard to mutations of highly conserved amino acids in glycoprotein hormone receptors.

• 한국임상약학회지
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• 제15권2호
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• pp.118-126
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• 2005

In our study, osteoporosis was induced by ovariectomized in female Sprague-Daweley (SD) rats, and the prevention and treatment efficacy of the climacteric disease the ostmenopausal type I pattern was examined by using the experimental substance soybeans and arowroot. No clinical signs and mortality after continuous oral administration of experimental substances for 14 weeks were not observed, however, the weight of experimental animals underwent ovariectomized was increased significantly in comparison with control group (p<0.01). The atrophy of the uterus due to T3 group was 23.64% after 7 weeks, 24.44% after 14 weeks (in comparison with N.C. group), and it was statistically significantly increased. In regard to the change of blood cells, it was observed that platelets were statistically significantly reduced in the ovariectomized group, and administered orally experimental substances continuously for 7 weeks, in all groups administered experimental substances, it was found that platelets had the tendency to be increased more than N. C. group. In regard to He change of blood biochemistry, removed the ovary, the concentration of ALP showed the tendency to be increased than control group, and particularly in T3 group, it was increased significantly. In regard to the concentration of cholesterol, in comparison with negative control group, it was reduced 68% at 7 weeks and 35% at 14 weeks. After ovariectomized, the amount of estrogen was found to be reduced by 21.37% in comparison with control group, it showed the tendency to be increased by 4.49% in T1 group and 7.62% in T2 group, the concentration of estrogen in each group showed the tendency to be increased than negative control group, and in T3 group, it was increased to 100.46% and 117.65% in T4 group, and it was increased more than control group. Based on the above experimental results, in the experimental animals female rats, because of the hormonal imbalance induced by ovariectomized, a large mount of fat is accumulated in the body and due to it osteoporosis, obesity, hypertension, hyperlipidemia, fat of the liver, arteriosclerosis, diabetes, and other metabolic diseases were developed. Hence, when the experimental substance Extraction of ferment arrowroot was orally administered continuously for 14 weeks, it was thought that a certain proportion of the hormonal balance was maintained that functioned as a substance interfering the accumulation of fat, and it was considered to be of help in the treatment of not only osteoporosis Type I, but also for the prevention and treatment of various endocrinal diseases.