• Title/Summary/Keyword: Oxidase

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Utilization of Ascidian, Halocynthia roretzi -4. Browning of Ascidian meat, Halocynthia roretzi and Its Prevention- (우렁쉥이 이용에 관한 연구 -4. 우렁쉥이 육의 갈변 및 그 방지-)

  • LEE Kang-Ho;CHO Ho-Sung;KIM Dong-Soo;HONG Byeong-Il;PARK Cheon-Soo;KIM Min-Gi
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.26 no.3
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    • pp.214-220
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    • 1993
  • Browning of ascidian, Halocynthia roretzi, meat occurres very rapidly when skinned off or cut during processing and it resulted the quality loss of fresh frozen, dehydrated or fermented products. In this study, the causes of color development and prevention of browning were experimented. The browning of ascidian meat may be occurred enzymatically by a tyrosinase contained in meat and viscera which acted specifically on L-tyrosine as a substrate rather than on catechol. Activity of the enzyme in viscera was three times higher than in meat. The optimum pH and temperature on the tyrosinase activity of crude enzyme obtained from ascidian was 6.0 and $30{\sim}35^{\circ}C$, respectively. The enzyme was inactivated heating at $80^{\circ}C$ for 3 minutes or $90{\sim}100^{\circ}C$ for 1 minute and it was inhibited by $0.1{\sim}0.5mM$ solutions at ascorbic acid, sodium hydrogen sulfite, cystein, citric acid, cyanide but only sodium hydrogen sulfite treatment was effective to retard such a high content of enzyme as in case of viscera. In practical use for processing of ascidian meat browning was retarded by dipping the viscera removed ascidian meat in 0.2M citric acid for 5 minutes or $0.2\%$ sodium hydrogen sulfite solution for 1 minute resulting in sulfur dioxide residue less than 100 ppm.

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Effects of Rhizobium Inoculation on the changes of Ureide-N and Amide-N Concentration in Stem and Root exudate of Soybean Plant (대두근류균(大豆根瘤菌) 접종(接種)이 뿌리와 줄기 즙액중(汁液中) Amide-N 및 Ureide-N 농도(濃度)에 미치는 영향(影響))

  • Ko, Jae-Young;Suh, Jang-Sun;Lee, Sang-Kyu
    • Korean Journal of Soil Science and Fertilizer
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    • v.22 no.4
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    • pp.329-336
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    • 1989
  • A series of green house experiment was conducted to find but the effect of fertilizer application and inoculation of rhizobium on the changes of amide-N, ureide-N and $NH_4-N$ concentration in stem and root exudates of soybean plant growth. The results obtained were summarized as follows ; 1. Five strains of indigenous Rhizobium japonicum-nitrogen fixing activity($C_2H_2$-reducing activity) was more than 6.4 to 20.1 nmole/hr/tube-were identified from 37 soil samples in 22 areas of farmers field throughout country. 2. These identified 5 strains of rhizobium were obtained high nitrate reductase but low ammonium and nitrite oxidase activities. Among 5 strains of rhizobium the Rhizobium japonicum RjK-134 was applied for this green house experiment. 3. Dry matter yield was increased by the combination of inoculation of Rhizobium japonicum RjK-134 with no fertilizer and without nitrogen fertilizer application. However, dry matter yield was decreased with application of N and NPK with inoculation of rhizobium. 4. The concentrations of amide-N and ureide-N were increased in xylem sap than that of root exudate and higher concentration was obtained ar 30 days after planting than flowering stage (45 days after planting). 5. The combination of NPK application with inoculation of Rhrizobium japonicum RjK-134 enhanced the increase of amide-N and ureide-N concentration in xylem sap and root exudate. 6. High ammonium-N concentration in xylem sap and root exudate were obtained in combination with without-fertilizer under no inoculation of rhizobium and N and NPK application with inoculation of rhizobium.

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Thrips Infesting Hot Pepper Cultured in Greenhouses and Variation in Gene Sequences Encoded in TSWV (시설재배지 고추를 가해하는 총채벌레류와 TSWV 유전자 서열 변이)

  • Kim, Chulyoung;Choi, Duyeol;Kang, Jeong Hun;Ahmed, Shabbir;Kil, Eui-Joon;Kwon, Gimyeon;Lee, Gwan-Seok;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.60 no.4
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    • pp.387-401
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    • 2021
  • Thrips infesting hot peppers were monitored in greenhouses using yellow sticky traps. In addition, the hot peppers infected with tomato spotted wilt virus (TSWV) were observed during the monitoring period. The flower thrips (Frankliniella intonsa) were initially trapped at a low density just after transplanting seedlings of hot peppers at late March. The western flower thrips (Frankliniella occidentalis) were trapped after mid April. These two thrips represented more than 98% of the total thrips attracted to the traps after May, in which F. intonsa showed higher occurrence frequency than F. occidentalis. The total number of thrips had two peaks at mid May with a small and short-term peak and at June-July with a large and long-term peak. The trapped thrips exhibited inconsistent sex ratios, suggesting a seasonal parthenogenesis. Different geographical populations were varied in cytochrome oxidase I sequences, in which local populations in Andong shared a high sequence similarity. TSWV-infected hot peppers, which might be mediated by these two thrips species, were observed and confirmed by an immunoassay kit and a molecular diagnosis using RT-PCR. In addition, the TSWV was detected in F. occidentalis collected from the infected hot peppers. Three open reading frames (NSS, N, and NSM) of the isolated TSWV genomes were sequenced and showed multiple point mutations containing missense mutations among geographical variants. When the isolated TSWV was fed to nonvirulent thrips of F. occidentalis, the virus was detected in both larvae and adults. However, the viral replication occurred in larvae, but not in adults.

Lysophosphatidic Acid Stimulates SKOV-3 Cell Migration through the Generation of Reactive Oxygen Species via the mTORC2/Akt1/NOX Signaling Axis (리소포스타티드산은 SKOV-3 난소암세포의 mTORC2/Akt1/NOX 신호전달 기전을 통해 활성산소를 형성하고 이를 통해 세포의 이동을 촉진)

  • Eun Kyoung Kim;Seo Yeon Jin;Jung Min Ha;Sun Sik Bae
    • Journal of Life Science
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    • v.33 no.2
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    • pp.129-137
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    • 2023
  • Reactive oxygen species (ROS) play an essential role in a variety of cellular physiological phenomena. The present study assessed the signaling axis that mediates the lysophosphatidic acid (LPA)-induced migration of SKOV-3 cells. Insulin-like growth factor-1 (IGF-1) stimulated SKOV-3 cell migration in a time- and dose-dependent manner. Similarly, LPA stimulated SKOV-3 cell migration and the phosphorylation of Akt in a time- and dose-dependent manner. The pharmacological inhibition of LPA receptors (LPA1/LPA3) significantly suppressed LPA-induced SKOV-3 cell migration. However, IGF-1-induced SKOV-3 cell migration was not affected by the inhibition of LPA1 and LPA3. Pharmacological inhibition of phosphoinositide 3-kinase (PI3K) or Rho-associated kinase (ROCK) significantly suppressed LPA-induced migration, whereas the inhibition of MAPK kinase (MEK) had no effect. Inhibition of PI3K or ROCK completely suppressed LPA-induced ROS generation, and suppression of nicotinamide adenine dinucleotide phosphate oxidase (NOX) or chelation of ROS by N-acetylcysteine (NAC) blocked LPA-induced SKOV-3 cell migration. LPA-induced ROS generation was suppressed by silencing Rictor or Akt1 but not Raptor or Akt2. Silencing Rictor or Akt1 significantly suppressed LPA-induced SKOV-3 cell migration, whereas silencing Raptor or Akt2 had no effect. Finally, the overexpression of the constitutively active form Akt1 (CA-Akt1) significantly enhanced the LPA-induced migration of SKOV-3 cells. Given these results, we suggest that LPA stimulates SKOV-3 cell migration by ROS generation, which is mediated by the mTORC2/Akt1/NOX signaling axis.

Loss of EMP2 Inhibits Melanogenesis of MNT1 Melanoma Cells via Regulation of TRP-2

  • Enkhtaivan, Enkhmend;Kim, Hyun Ji;Kim, Boram;Byun, Hyung Jung;Yu, Lu;Nguyen, Tuan Minh;Nguyen, Thi Ha;Do, Phuong Anh;Kim, Eun Ji;Kim, Kyung Sung;Huy, Hieu Phung;Rahman, Mostafizur;Jang, Ji Yun;Rho, Seung Bae;Lee, Ho;Kang, Gyeoung Jin;Park, Mi Kyung;Kim, Nan-Hyung;Choi, Chang Ick;Lee, Kyeong;Han, Hyo Kyung;Cho, Jungsook;Lee, Ai Young;Lee, Chang Hoon
    • Biomolecules & Therapeutics
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    • v.30 no.2
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    • pp.203-211
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    • 2022
  • Melanogenesis is the production of melanin from tyrosine by a series of enzyme-catalyzed reactions, in which tyrosinase and DOPA oxidase play key roles. The melanin content in the skin determines skin pigmentation. Abnormalities in skin pigmentation lead to various skin pigmentation disorders. Recent research has shown that the expression of EMP2 is much lower in melanoma than in normal melanocytes, but its role in melanogenesis has not yet been elucidated. Therefore, we investigated the role of EMP2 in the melanogenesis of MNT1 human melanoma cells. We examined TRP-1, TRP-2, and TYR expression levels during melanogenesis in MNT1 melanoma cells by gene silencing of EMP2. Western blot and RT-PCR results confirmed that the expression levels of TYR and TRP-2 were decreased when EMP2 expression was knocked down by EMP2 siRNA in MNT1 cells, and these changes were reversed when EMP2 was overexpressed. We verified the EMP2 gene was knocked out of the cell line (EMP2 CRISPR/Cas9) by using a CRISPR/Cas9 system and found that the expression levels of TRP-2 and TYR were significantly lower in the EMP2 CRISPR/Cas9 cell lines. Loss of EMP2 also reduced migration and invasion of MNT1 melanoma cells. In addition, the melanosome transfer from the melanocytes to keratinocytes in the EMP2 KO cells cocultured with keratinocytes was reduced compared to the cells in the control coculture group. In conclusion, these results suggest that EMP2 is involved in melanogenesis via the regulation of TRP-2 expression.

Overcoming taxonomic challenges in DNA barcoding for improvement of identification and preservation of clariid catfish species

  • Piangjai Chalermwong;Thitipong Panthum;Pish Wattanadilokcahtkun;Nattakan Ariyaraphong;Thanyapat Thong;Phanitada Srikampa;Worapong Singchat;Syed Farhan Ahmad;Kantika Noito;Ryan Rasoarahona;Artem Lisachov;Hina Ali;Ekaphan Kraichak;Narongrit Muangmai;Satid Chatchaiphan6;Kednapat Sriphairoj;Sittichai Hatachote;Aingorn Chaiyes;Chatchawan Jantasuriyarat;Visarut Chailertlit;Warong Suksavate;Jumaporn Sonongbua;Witsanu Srimai;Sunchai Payungporn;Kyudong Han;Agostinho Antunes;Prapansak Srisapoome;Akihiko Koga;Prateep Duengkae;Yoichi Matsuda;Uthairat Na-Nakorn;Kornsorn Srikulnath
    • Genomics & Informatics
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    • v.21 no.3
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    • pp.39.1-39.15
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    • 2023
  • DNA barcoding without assessing reliability and validity causes taxonomic errors of species identification, which is responsible for disruptions of their conservation and aquaculture industry. Although DNA barcoding facilitates molecular identification and phylogenetic analysis of species, its availability in clariid catfish lineage remains uncertain. In this study, DNA barcoding was developed and validated for clariid catfish. 2,970 barcode sequences from mitochondrial cytochrome c oxidase I (COI) and cytochrome b (Cytb) genes and D-loop sequences were analyzed for 37 clariid catfish species. The highest intraspecific nearest neighbor distances were 85.47%, 98.03%, and 89.10% for COI, Cytb, and D-loop sequences, respectively. This suggests that the Cytb gene is the most appropriate for identifying clariid catfish and can serve as a standard region for DNA barcoding. A positive barcoding gap between interspecific and intraspecific sequence divergence was observed in the Cytb dataset but not in the COI and D-loop datasets. Intraspecific variation was typically less than 4.4%, whereas interspecific variation was generally more than 66.9%. However, a species complex was detected in walking catfish and significant intraspecific sequence divergence was observed in North African catfish. These findings suggest the need to focus on developing a DNA barcoding system for classifying clariid catfish properly and to validate its efficacy for a wider range of clariid catfish. With an enriched database of multiple sequences from a target species and its genus, species identification can be more accurate and biodiversity assessment of the species can be facilitated.

L-AHG-mediated Suppression of M1 Polarization and Pro-inflammatory Signaling Pathways in LPS-stimulated RAW264.7 Macrophages (LPS에 의해 자극된 RAW264.7 대식세포에서 L-AHG에 의한 M1 분극화 및 친염증 신호 경로의 억제)

  • Won Young Jang;Shin Young Park;Ki Youn Kim;Do Youn Jun;Young-Seuk Bae;Young Ho Kim
    • Journal of Life Science
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    • v.34 no.7
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    • pp.443-452
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    • 2024
  • This study aimed to examine the influence of 3,6-anhydroxygalactose (L-AHG) on the pro-inflammatory M1 polarization and pro-inflammatory responses observed in the RAW264.7 mouse macrophage cell line following stimulation with lipopolysaccharides (LPS). L-AHG exhibited a significant and dose-dependent inhibition of inducible nitric oxide synthase (iNOS) expression, a hallmark of M1 polarization, and subsequent NO production in LPS-stimulated RAW264.7 cells. Furthermore, the LPS-induced upregulation of cyclooxygenase-2 (COX-2), which drives the production of prostaglandin E2, an inflammatory mediator, was also inhibited by L-AHG. L-AHG did not affect the LPS-triggered Toll-like receptor 4 (TLR4)-mediated pro-inflammatory signaling pathway, which culminated in the activation of transforming growth factor-β-activated kinase 1 (TAK1). However, it was observed to inhibit the generation of reactive oxugen species (ROS) in a dose-dependent manner, as well as the TAK1-driven activation of JNK and p38 MAPK. Given that the active p38 MAPK is known to contribute to the assembly of active nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, which catalyzes the intracellular generation of pro-inflammatory ROS in LPS-stimulated macrophages, the dose-dependent reduction in the LPS-induced ROS generation by L-AHG may be mainly due to the prevention of TAK1-driven activation of p38 MAPK. Together, these results demonstrate that the L-AHG-mediated inhibition of the TAK1-JNK/p38 MAPK activation phase of the pro-inflammatory signaling pathway in LPS-stimulated RAW264.7 cells by L-AHG represents a promising mechanism for suppressing M1 polarization and pro-inflammatory responses in macrophages.

Effects of oxypeucedanin hydrate isolated from Angelica dahurica on myoblast differentiation in association with mitochondrial function (백지에서 추출한 oxypeucedanin hydrate의 미토콘드리아 기능 관련 근생성 효과)

  • Eun-Ju Song;Ji-Won Heo;Jee Hee Jang;Yoon-Ju Kwon;Yun Hee Jeong;Min Jung Kim;Sung-Eun Kim
    • Journal of Nutrition and Health
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    • v.57 no.1
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    • pp.53-64
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    • 2024
  • Purpose: Mitochondria play a crucial role in preserving skeletal muscle mass, and damage to mitochondria leads to muscle mass loss. This study investigated the effects of oxypeucedanin hydrate, a furanocoumarin isolated from Angelica dahurica radix, on myogenesis and mitochondrial function in vitro and in zebrafish models. Methods: C2C12 myotubes cultured in media containing 0.1, 1, 10, or 100 ng/mL oxypeucedanin hydrate were immunostained with myosin heavy chain (MHC), and then multinucleated MHC-positive cells were counted. The expressions of markers related to muscle differentiation, muscle protein degradation, and mitochondrial function were determined by quantitative reverse transcription polymerase chain reaction. To investigate the effects of oxypeucedanin hydrate on mitochondrial dysfunction, Tg(Xla.Eef1a1:mito-EGFP) zebrafish embryos were treated with 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI) with or without oxypeucedanin hydrate and analyzed for mito-EGFP intensity and mitochondrial length. Results: Oxypeucedanin hydrate significantly increased MHC-positive multinucleated myotubes (≥ 3 nuclei) and increased the expression of the myogenic marker myosin heavy chain 4. However, it decreased the expressions of muscle-specific RING finger protein 1 and muscle atrophy f-box (markers of muscle protein degradation). Furthermore, oxypeucedanin hydrate enhanced the expressions of markers of mitochondrial biogenesis (peroxisome proliferator-activated receptor-gamma coactivator 1 alpha, transcription factor a mitochondrial, succinate dehydrogenase complex flavoprotein subunit A, and cytochrome c oxidase subunit 1) and mitochondrial fusion (optic atrophy 1). However, it reduced the expression of dynamin-related protein 1 (a mitochondrial fission regulator). Consistently, oxypeucedanin hydrate reduced FOLFIRI-induced mitochondrial dysfunction in the skeletal muscles of zebrafish embryos. Conclusion: The study indicates that oxypeucedanin hydrate promotes myogenesis by improving mitochondrial function, and thus, suggests oxypeucedanin hydrate has potential use as a nutritional supplement that improves muscle mass and function.

Estimation of Adult Season of Occurrence and Annual Generation Numbers of the Asiatic Pink Stem Borer, Sesamia inferens (Walker, 1856) (Lepidoptera: Noctuidae) (벼밤나방(Sesamia inferens (Walker, 1856))(나비목: 밤나방과) 성충 발생시기와 연중 세대수 추정)

  • Eun Young Kim;Young-Mi Park;Soon Do Bae;Gwan-Seok Lee;Chae-Hoon Paik;Do-Ik Kim;Wonhoon Lee;Jin Kyo Jung;Bo Yoon Seo
    • Korean journal of applied entomology
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    • v.62 no.4
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    • pp.227-243
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    • 2023
  • The Asiatic pink stem borer, Sesamia inferens (Walker, 1856) (Lepidoptera: Noctuidae) is a pest that attacks rice stems. In this study, we estimated the annual generation of insect in several regions of the Republic of Korea. Adult trapping using a sex pheromone trap detected the occurrence of S. inferens adults in the three northernmost areas around 38° latitude and showed that the insect inhabits all of Korea. In most areas investigated, the seasons of the adult generations estimated using the single-sine degree-day model did not deviate from the corresponding observed seasons of adult occurrence. We estimated that the overwintering larvae hypothetically-estimated using the model could be originated from the last generation of adults. When larvae collected in paddy fields during the autumn season in a few middle and southern areas were reared at 25℃, ca. 70% of pupae did not show additional larval molting before their pupation. All larvae collected in early March in a southern area (Goseong, Gyeongsangnam-do) pupated without additional larval molting when reared at 25℃. Based on these results, we presumed that S. inferens could overwinter as mainly the last instar larval stage in the area. Taken together, we conclude that S. inferens primarily has two generations per year in areas around 38° latitude, and three generations in the areas between 35.3° and 37.3° latitude. In addition, approximately 35% of insects captured by the sex pheromone trap were species other than S. inferens, as determined by analyzing the nucleotide sequences of the cytochrome c oxidase 1 gene. These species were not morphologically misidentified as S. inferens.

Inhibitory Effects of Amitriptyline, Sertraline and Chlorpromazine on the Thrombin-induced Aggregation of Platelets (Thrombin성 혈소판응집에 대한 Amitriptyline, Sertraline 및 Chlorpromazine의 억제작용)

  • Choi, Sang-Hyun;Lee, Young-Jae;Shin, Kyung-Ho;Chun, Yeon-Sook;Chun, Boe-Gwun
    • The Korean Journal of Pharmacology
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    • v.31 no.3
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    • pp.299-311
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    • 1995
  • Platelets resemble monoaminergic neurons in several respects, i.e. the uptake of 5-HT and its inhibition, the subcellular storage and release of 5-HT, and the metabolism of aromatic amines brought about by monoamine oxidase. And the 5-HT content of rabbit platelets is well known to be about 40 times higher than that of human platelets. Therefore, this study was carried out to investigate the influences of amitriptyline (AMT) and sertraline (SRT) on the aggregation, contents of signaling second messengers, and protein phosphorylations of rabbit platelets in response to thrombin, 0.25 unit/ml, comparing with those of chlorpromazine (CPZ). Thrombin-induced aggregation was inhibited by SRT $(IC50:4.37{\times}10^{-5}\;M)$, CPZ $(IC50:5.76{\times}10^{-5}\;M)$, and AMT $(IC50:1.15{\times}10^{-4}\;M)$, respectively, and the aggregation by A23187 $(1.0\;{\mu}M)$ or PMA (320 nM) was also inhibited by SRT, CPZ, and AMT. AMT, SRT, and CPZ had little affects on basal contents of platelet $TXB_2$ and $PGE_2$, but all of them inhibited the thrombin-induced increase of $TXB_2$. Thrombin did not change the platelet contents of cAMP and cGMP. CPZ, AMT, and SRT produced the slight decrease of basal cAMP content, and their effects were not affected by thrombin-treatment. But SRT and AMT moderately increased the basal cGMP content, and the cGMP content of thrombin-stimulated platelets was gradually increased by the pretreatment with SRT, AMT, and CPZ. Particularly, the SRT-dependent increase of the cGMP content was notable. Platelet $Ins(1,4,5)P_3$ content was rapidly increased up to a plateau within 10 sec after thrombin-stimulation, AMT, SRT, and CPZ increased the basal $Ins(1,4,5)P_3$ content, and the thrombin-dependent increase was enhanced by pretreatment with CPZ and AMT, but was blunted by SRT. Platelet $[Ca^{2+}]_i$, was rapidly increased up to a peak level within 20 sec after thrombin-stimulation. The increase of $[Ca^{2+}]_i$ was sisnificantly inhibited by AMT, SRT, and CPZ. Thrombin- or PMA-induced phosphorylations of platelet $41{\sim}43\;kDa$ and 20 kDa proteins were significantly inhibited by AMT, SRT, and CPZ. These results suggest that the antiplatelet activities of AMT and CPZ may be considerably attributed to the inhibition of protein kinase C activity, and the activity of SRT may be associated with the inhibitory effect on the thrombin-induced increase of $Ins(1,4,5)P_3$ and the increasing effect on the cGMP content of ptatelets. Therefore, it seems to be evident that AMT and SRT may produce their antidepressant activity, at least, partly through the inhibition of protein kinase C activity or the increase of resting $Ins(1,4,5)P_3$, content and in case of SRT, to a lesser extent, via the increase of cGMP in the brain.

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