• Title/Summary/Keyword: primary metabolism

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Acid Tolerance Response of Streptococcus mutans at Anaerobic Condition (Streptococcus mutans의 혐기적 산 내성도 평가)

  • Han, Yang-Keum;Song, Sang-Sun;Lee, In-Soo
    • Journal of dental hygiene science
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    • v.1 no.1
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    • pp.7-11
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    • 2001
  • Streptococcus mutans is one of the primary bacteria that cause dental caries which further result in plaque build up. Acid production resulted from carbohydrate metabolism can threaten survival of the bacteria. However some populations of S. mutans which are exposed to low acidic condition for a period of time would develop resistance and tolerance of cells to acidity that will enhance the chance of survival. Similar acid tolerances has been reported in case of Salmonella enterica serovar typhimurium, E. coli, Shigella flexneri. These acid tolerance responses(ATR) have been evolved in a similar manner as S. mutans. The protein produced in acidic condition has been proven to be important for ATR and confirmed by using chloramphenicol procedure. We hypothesize here that proteins synthesized in response to acid shock and other elements are important for ATR of cells. In this study we have confirmed that S. mutans developed acid tolerance and resistance against anaerobic condition. Mutational DNA analysis responsible for acid tolerance should be additionally required in the future. Since the development of acid tolerance that is essential for the survival of S. mutans and development of dental caries, ATR of S. mutans shoule be farther to prevent dental caries.

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Effects of Compound K on Insulin Secretion and Carbohydrate Metabolism (Compound K의 인슐린분비 및 탄수화물 대사에 미치는 영향)

  • Choi, Yun-Suk;Han, Gi-Cheol;Han, Eun-Jung;Park, Kum-Ju;Sung, Jong-Hwan;Chung, Sung-Hyun
    • Journal of Ginseng Research
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    • v.31 no.2
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    • pp.79-85
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    • 2007
  • Compound K (CK) is a final metabolite of panaxadiol ginsenosides. Although panax ginseng is known to have anti-diabetic activity, the active ingredient is not yet fully identified. Therefore, it would be interesting to know whether and how CK has an anti-diabetic activity. First, insulin secretion-stimulating activity of CK was examined using RIN-m5F cell line and primary cultured islets. CK enhanced the insulin secretion in a concentration dependent manner. This effect, however, was almost completely abolished in the presence of diazoxide, $K^+$ channel opener, indicating that the insulin secretion-stimulating activity of CK is presumably due to blockade of ATP sensitive $K^+$ channel. In addition, effects of CK on gene expressions of hepatic enzymes (phosphoenolpyruvate carboxykinase[PEPCK], glucose-6-phos-phatase[G6Pase]) and on adipocyte differentiation in H4IIE and 3T3-Ll cells, respectively, were examined. CK suppressed the induction of PEPCK and G6Pase mRNA expressions under the dexamethasone/cAMP stimulation condition. CK also reduced the $PPAR-{\gamma}$ mRNA expression and triglyceride accumulation in a dose dependent manner as compared to the control. The present study suggests that CK deserves to examine whether it shows an anti-diabetic activity in animal and human studies.

Cloning and Functional Analysis of Gene Coding for S-Adenosyl-L-Methionine Synthetase from Streptomyces natalensis (Streptomyces natalensis로부터 S-adenosyl-L-methionine synthetase 유전자의 클로닝 및 기능분석)

  • Yoo, Dong-Min;Hwang, Yong-Il;Choi, Sun-Uk
    • Journal of Life Science
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    • v.21 no.1
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    • pp.96-101
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    • 2011
  • S-Adenosyl-L-methionine synthtase (SAM-s) catalyzes the biosynthesis of SAM from ATP and L-methionine. SAM plays important roles in the primary and secondary metabolism of cells. A metK encoding a SAM-s was searched from Streptomyces natalensis producing natamycin, a predominantly a strong antifungal agent, inhibiting the growth of both yeasts and molds and preventing the formation of aflatoxin in filamentous fungi. To obtain the metK of S. natalensis, PCR using primers designed from the two highly conserved regions for metK genes of Streptomyces strains was carried out, and an intact 1.2-kb metK gene of S. natalensis was cloned by genomic Southern hybridization with PCR product as a probe. To identify the function of the cloned metK gene, it was inserted into pSET152ET for its high expression in the Streptomyces strain, and then introduced into S. lividans TK24 as a host by transconjugation using E. coli ET12567(pUZ8002). The high expression of metK in S. lividans TK24 induced actinorhodin production on R5 solid medium, and its amount in R4 liquid medium was 10-fold higher than that by exconjugant including only pSET152ET.

Regulation of hPTH Expression In Virto Using the Tetracycline Inducible Retrovirus Vector System (Tetracycline Inducible Retrovirus Vector System을 이용한 In Vitro에서의 인간 부갑상선 호르몬의 발현 조절)

  • Koo, Bon-Chul;Kwon, Mo-Sun;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.30 no.3
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    • pp.157-162
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    • 2006
  • Endogenous 84 amino acid parathyroid hormone(PTH) is synthesized as a pre-pro hormone by the chief cells of the parathyroid glands. Physiological actions of PTH include regulation of bone metabolism, renal tubular reabsorption of calcium and phosphate, and intestinal calcium absorption. In addition, PTH stimulates new bone formation by extraordinary stimulation of osteoblastic activity and decreasing calcium excretion by the kidney. In this study, we constructed and tested retrovirus vectors designed to express the human parathyroid hormone(hPTH) gene under the control of the tetracycline-inducible promoters. To increase the hPTH gene expression at turn-on state, woodchuck hepatitis virus posttranscriptional regulatory element(WPRE) sequence was also introduced into retrovirus vector at downstream region of either the hPTH gene or the sequence encoding reverse tetracycline-controlled transactivator(rtTA). Transformed primary culture cells(porcine fetal fibroblast, PFF, chicken embryonic fibroblast, CEF) were cultured in the medium supplemented with or without doxycycline(tetracycline derivative) for 48 hours, and induction efficiency was measured by comparing the hPTH gene expression level using two step RT-PCR and ELISA Higher hPTH expression($3{\tims}10^4\;pg/ml,\;5.3{\times}10^4\;pg/ml$) and tighter expression control(up to 8 fold) were observed from the vector in which the WPRE sequence was placed at downstream of the hPTH gene. The resulting tetracycline inducible vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals.

Randomized Double-blind Human Trial to Evaluate Efficacy and Safety of Rhus verniciflua Stokes (Lacca Sinica Exsiccata) and Eucommia ulmoides Oliver (Eucommiae Cortex) Extract Combination (ILF-RE) on Improvement of Liver Function (옻나무-두충추출혼합물(ILF-RE)의 간기능 개선에 대한 유효성 및 안전성을 평가하기 위한 무작위배정 이중눈가림 인체적용시험)

  • Youn, Young;Baek, Hyang-Im;Jin, Hee-Yeon;Jeong, Da-Young;Shen, Lei;Joo, Jong-Cheon;Park, Soo-Jung
    • The Korea Journal of Herbology
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    • v.35 no.1
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    • pp.45-55
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    • 2020
  • Objectives : The purpose of this study is to determine whether Rhus verniciflua Stokes with Latin name Lacca Sinica Exsiccata, and Eucommia ulmoides Oliver with Latin name Eucommiae Cortex Extract Combination (ILF-RE) improves laboratory test results in participants with liver function disorder. Methods : This study was conducted at Woosuk university Korean medicine hospital where participants with high serum alanine transaminase (ALT) levels from 45 to 135 U/L were enrolled. Subjects received ILF-RE 3.6 g (1.2 g/day as ILF-RE) or placebo 3.6 g for 12 weeks. It was confirmed that urushiol was not detected in ILF-RE. The primary outcomes were the decrement degree of serum ALT and gamma-glutamyl transferase (GGT) levels between two groups. The secondary outcomes were the decrement degree of serum aspartate transaminase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LD), total bilirubin, total cholesterol, triglyceride (TG) and fatty liver index (FLI) levels between two groups. Adverse events, skin prick tests, laboratory tests, and vital signs were observed and analyzed to confirm the safety of ILF-RE.1) Results : In the ILF-RE group, the liver function index ALT, GGT, lipid metabolism index TG, and fatty liver index FLI were significantly decreased compared to the placebo group. There was no significant difference in ILF-RE group in terms of adverse events, severe adverse events, skin prick test, laboratory test, and vital signs compared with placebo group. Conclusions : ILF-RE was found to be effective in improving liver function. In addition, no clinically significant adverse events or body changes were observed during this study.

MicroRNA analysis reveals the role of miR-214 in duck adipocyte differentiation

  • Wang, Laidi;Hu, Xiaodan;Wang, Shasha;Yuan, Chunyou;Wang, Zhixiu;Chang, Guobin;Chen, Guohong
    • Animal Bioscience
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    • v.35 no.9
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    • pp.1327-1339
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    • 2022
  • Objective: Fat deposition in poultry is an important factor in production performance and meat quality research. miRNAs also play important roles in regulating adipocyte differentiation process. This study was to investigate the expression patterns of miRNAs in duck adipocytes after differentiation and explore the role of miR-214 in regulating carnitine palmitoyltransferases 2 (CPT2) gene expression during duck adipocyte differentiation. Methods: Successful systems for the isolation, culture, and induction of duck primary fat cells was developed in the experiment. Using Illumina next-generation sequencing, the miRNAs libraries of duck adipocytes were established. miRanda was used to predict differentially expressed (DE) miRNAs and their target genes. The expression patterns of miR-214 and CPT2 during the differentiation were verified by quantitative real-time polymerase chain reaction and western blot. Luciferase reporter assays were used to explore the specific regions of CPT2 targeted by miR-214. We used a miR-214 over-expression strategy in vitro to further investigate its effect on differentiation process and CPT2 gene transcription. Results: There were 481 miRNAs identified in duck adipocytes, included 57 DE miRNA candidates. And the 1,046 targets genes of DE miRNAs were mainly involved in p53 signaling, FoxO signaling, and fatty acid metabolism pathways. miR-214 and CPT2 showed contrasting expression patterns before and after differentiation, and they were selected for further research. The expression of miR-214 was decreased during the first 3 days of duck adipocytes differentiation, and then increased, while the expression of CPT2 increased both in the transcriptional and protein level. The luciferase assay suggested that miR-214 targets the 3'untranslated region of CPT2. Overexpression of miR-214 not only promoted the formation of lipid droplets but also decreased the protein abundance of CPT2. Conclusion: Current study reports the expression profile of miRNAs in duck adipocytes differentiated for 4 days. And miR-214 has been proved to have the regulator potential for fat deposition in duck.

Effect of Lythrum salicaria Extract on Body Fat Reduction: A Protocol for a Randomized, Double-Blinded, Placebo-Controlled Clinical Trial (체지방 감소에 대한 털부처꽃 추출물의 효과: 무작위배정, 이중눈가림, 대조군 비교 인체적용시험 프로토콜)

  • Hye-Jin Park;In Heo;Yea-Jin Park;Hyo-Jin An;Su Shin;Yun-Yeop Cha
    • Journal of Korean Medicine for Obesity Research
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    • v.24 no.1
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    • pp.87-93
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    • 2024
  • Objectives: Obesity is a globally prevalent public health issue. Hence, there is a need for the development of safer and more effective anti-obesity drugs. Lythrum salicaria, a traditional medicinal herb used for centuries, has been reported to improve lipid metabolism and fat accumulation. It also has a low toxicity profile. Therefore, its potential as a functional ingredient in health functional foods needs to be evaluated. Methods: In this randomized, double-blind, placebo-controlled clinical trial, 90 participants will be randomly assigned to either the experimental or control group. Each subject will orally receive L. salicaria extract (1,350 mg/day) (500 mg L. salicaria+850 mg lactose as vehicle) or lactose (1,350 mg/day) as a hard capsule formula for 84 days (12 weeks). The primary outcome will be body fat mass (kg), which will be assessed using dual-energy x-ray absorptiometry (DXA) (performed only at visits 2 and 4). Secondary outcomes include body mass index, body weight, waist-to-hip ratio, body fat percentage (%) measured using DXA, lean body mass (kg) measured using DXA (assessed only at visits 2 and 4), lipids (total cholesterol, triglyceride, high-density lipoprotein cholesterol, and calculated low-density lipoprotein cholesterol), free fatty acid, high sensitivity C-reactive protein, adiponectin, and leptin. Conclusions: This protocol will be implemented after approval of Institutional Review Board of Pusan National University Korean Medicine Hospital (approval number: PNUKHIRB-2022-08-002) and registration with the Korean National Clinical Research Information Service (CRIS) (CRIS-KCT0008060). The results of this trial will provide potential of L. salicaria as a new anti-obesity functional food with fat-reducing effects and low toxicity.

Ecosysteme de I′Etang de Berre (Mediterranee nord-occidentale) : Caracteres Generales Physiques, Chimiques et Biologiques

  • Kim, Ki-Tai
    • Korean Journal of Environmental Biology
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    • v.22 no.2
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    • pp.247-258
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    • 2004
  • Climatological, hydrological and planktonical research studies, measurements of primary production and photosynthetic efficiency from December 1976 to December 1978 have been carried out in two brackish lakes: Lake Etang de Berre and Lake Etang de Vaine located in the French Mediterranean coast, in the region of Carry-le-Rouet located on the north-west Mediterranean near Marseilles, and in fresh water inflows from 4 Rivers (Touloubre, Durance, Arc, Durancole) to Lake Etang de Berre. Physico-chemical parameters were measured for this study: water temperature, salinity, density, pH, alcalinity, dissolved oxygen (% saturation), phosphate, nitrate, nitrite, silicate etc. Diverse biological parameters were also studied: photosynthetic pigments, phaeopigments, specific composition and biomass of phytoplankton, primary pelagic production etc. Climatical factors were studied: air-temperature, solar-radiation, evaporation, direction (including strength) of winds, precipitation and freshwater volume of the four rivers. The changes in Lake ‘Etang de Berre’ ecosystem depend on the quality of the water in the Durance River, and on the effects of seawater near the entrance of the Caronte Canal. The water quality of the lake varies horizontally and vertically as a result of atmospheric phenomena, maritime currents and tides. The distribution of water temperatures is generally heterogeneous. Southeasterly winds and the Northeasterly Mistral wind are important in the origins of circulated and mixed water masses. These winds are both frequent and strong. They have, as a result, a great effect on the water environment of Lake Etang de Berre. In theory, the annual precipitation in this region is well over eight times the water mass of the lake. The water of the Durance River flows into Lake Etang de Berre through the EDF Canal, amounting to 90% of the precipitation. However, reduction of rainfall in dry seasons has a serious effect on the hydrological characteristics of the lake. The temperature in the winter is partially caused by the low temperature of fresh water, particularly that of the Durance River. The hydrological season of fresh and brackish water is about one month ahead of the hydrological season of sea water in its vicinity. The salinity of Lake Etang de Berre runs approximately 3$\textperthousand$, except at lower levels and near the entrance to the Caronte Canal. However, when the volume of the Durance River water is reduced in the summer and fall, the salinity rises to 15$\textperthousand$. In the lake, the ratio of fresh water to sea water is six to one (6:1). The large quantities of seston conveyed by rivers, particularly the Durance diversion, strongly reduce the transparency in the brackish waters. Although the amount of sunshine is also notable, transparency is slight because of the large amount of seston, carried chiefly by Tripton in the fresh water of the Durance River. Therefore, photosynthesis generally occurs only in the surface layer. The transparency progressively increases from freshwater to open seawater, as mineral particles sink to the bottom (about 1.7kg $m^{-2}a^{-1}$ on the average in brackish lakes). The concentration of dissolved oxygen and the rate of oxygen saturation in seawater (Carry-le-Rouet) ranged from 5.0 to 6.0 $m\ell$ㆍ.$1^{-1}$, and from 95 to 105%, respectively. The amount of dissolved oxygen in Etang de Berre oscillated between 2.9 and 268.3%. The monographs of phosphate, nitrate, nitrite and silicate were published as a part of a study on the ecology of phytoplankton in these environments. Horizontal and vertical distributions of these nutriments were studied in detail. The recent diversion of the Durance River into Lake Etang de Berre has effected a fundamental change in this formerly marine environment, which has had a great impact in its plankton populations. A total of 182 taxa were identified, including 111 Bacillariophyceae, 44 Chlorophyceae, and 15 Cyanophyceae. The most abundant species are small freshwater algae, mainly Chlorophyceae. The average density is about $10^{8}$ cells $1^{-1}$ in Lake Etang de Berre, and about double that amount in Lake Etang de Vaine. Differences in phytoplankton abundance and composition at the various stations or at various depths are slight. Cell biovolume V (equivalent to true biomass), plasma volume VP (‘useful’ biomass) and, simultaneously. the cell surface area S and S/V ratio through the measurement of cell dimensions were computed as the parameters of phytoplankton productivity and metabolism. Pigment concentrations are generally very high on account of phytoplankton blooms by Cyanophyceae, Chlorophyceae and Cryptophyceae. On the other hand, in freshwaters and marine waters, pigment concentrations are comparatively low and stable, showing slight annual variation. The variations of ATP concentration were closely related to those of chlorophyll a and phytoplankton blooms only in marine waters. The carbon uptake rates ranged between 38 and 1091 mg$Cm^{-2}d^{-1}$, with an average surface value of 256 mg; water-column carbon-uptake rates ranged between 240 and 2310 mg$Cm^{-2}d^{-1}$, with an average of 810, representing 290 mg$Cm^{-2}$, per year 45 000 tons per year of photosynthetized carbon for the whole lake. Gross photosynthetic production measured by the method of Ryther was studied over a 2-year period. The values obtained from marine water(Carry-le-Rouet) ranged from 23 to 2 337 mg$Cm^{-2}d^{-1}$, with a weighted average of 319, representing about 110 gCm$^{-2}$ per year. The values in brakish water (Etang de Berre) ranged from 14 to 1778 mg$Cm^{-2}d^{-1}$, with a weighted average of 682, representing 250 mg$Cm^{-2}$ per year and 38 400 tons per year of photosynthesized carbon for the whole lake.

Diagnostic Efficacy of FDG-PET Imaging in Solitary Pulmonary Nodule (고립성폐결절의 진단시 FDG-PET의 임상적 유용성에 관한 연구)

  • Cheon, Eun Mee;Kim, Byung-Tae;Kwon, O. Jung;Kim, Hojoong;Chung, Man Pyo;Rhee, Chong H.;Han, Yong Chol;Lee, Kyung Soo;Shim, Young Mog;Kim, Jhingook;Han, Jungho
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.6
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    • pp.882-893
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    • 1996
  • Background : Over one-third of solitary pulmonary nodules are malignant, but most malignant SPNs are in the early stages at diagnosis and can be cured by surgical removal. Therefore, early diagnosis of malignant SPN is essential for the lifesaving of the patient. The incidence of pulmonary tuberculosis in Korea is somewhat higher than those of other countries and a large number of SPNs are found to be tuberculoma. Most primary physicians tend to regard newly detected solitary pulmonary nodule as tuberculoma with only noninvasive imaging such as CT and they prefer clinical observation if the findings suggest benignancy without further invasive procedures. Many kinds of noninvasive procedures for confirmatory diagnosis have been introduced to differentiate malignant SPNs from benign ones, but none of them has been satisfactory. FOG-PET is a unique tool for imaging and quantifying the status of glucose metabolism. On the basis that glucose metabolism is increased in the malignant transfomled cells compared with normal cells, FDG-PET is considered to be the satisfactory noninvasive procedure which can differentiate malignant SPNs from benign SPNs. So we performed FOG-PET in patients with solitary pulmonary nodule and evaluated the diagnostic accuracy in the diagnosis of malignant SPNs. Method : 34 patients with a solitary pulmonary nodule less than 6 cm of irs diameter who visited Samsung Medical Center from Semptember, 1994 to Semptember, 1995 were evaluated prospectively. Simple chest roentgenography, chest computer tomography, FOG-PET scan were performed for all patients. The results of FOG-PET were evaluated comparing with the results of final diagnosis confirmed by sputum study, PCNA, fiberoptic bronchoscopy, or thoracotomy. Results : (I) There was no significant difference in nodule size between malignant (3.1 1.5cm) and benign nodule(2.81.0cm)(p>0.05). (2) Peal SUV(standardized uptake value) of malignant nodules (6.93.7) was significantly higher than peak SUV of benign nodules(2.71.7) and time-activity curves showed continuous increase in malignant nodules. (3) Three false negative cases were found among eighteen malignant nodule by the FDG-PET imaging study and all three cases were nonmucinous bronchioloalveolar carcinoma less than 2 em diameter. (4) FOG-PET imaging resulted in 83% sensitivity, 100% specificity, 100% positive predictive value and 84% negative predictive value. Conclusion: FOG-PET imaging is a new noninvasive diagnostic method of solitary pulmonary nodule thai has a high accuracy of differential diagnosis between malignant and benign nodule. FDG-PET imaging could be used for the differential diagnosis of SPN which is not properly diagnosed with conventional methods before thoracotomy. Considering the high accuracy of FDG-PET imaging, this procedure may play an important role in making the dicision to perform thoracotomy in diffcult cases.

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Serotonin Synthesis and Metabolism in Dissociated Cultures of Fetal Rat Brainstem (흰쥐 태아 뇌간의 일차 세포배양에서 Serotonin의 합성 및 대사에 대한 연구)

  • Kim, Yung-Hi;Song, Dong-Keun;Wie, Myung-Bok;Song, Joon-Ho;Choi, Yeun-Sik
    • The Korean Journal of Pharmacology
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    • v.26 no.1
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    • pp.1-6
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    • 1990
  • We established an in vitro system of central serotonergic neurons by culturing dissociated rat embryonic (El4) brainstem cells to 14 days in vitro and monitored the serotonergic neuronal growth by measuring 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) contents in the cells with hish performance liquid chromatography with electrochemical detection (HPLC-EC). We studied also tile effects of various drugs on the contents of 5-HT and 5-HIAA, confirming in vivo reports. The 5-HT content (13 ng/mg protein) and 5-HT turnover rate (17 pmol/mg protein/h) at 14 days in vitro were in good agreement with those reported in the adult rat brain. The 5-HT content was more easily depleted with p-chlorophenylalanine, a tryptophan hydroxylase inhibitor than with NSD 1015 (3-hydroxybenzylhydrazine), an aromatic L-amino acid decarboxylase (AADC) inhibitor. Incubation of the cultures with tryptophan or 5-hydroxytryptophan increased the rate of serotonin formation implying that neither tryptophan hydroxylase nor AADC is saturated with its amino acid substrate in this in vitro system . The 5-HT content was depleted by reserpine. The 5-HT and 5-HIAA contents were increased and decreased, respectively, by monoamine oxidase inhibitors. All the above results indicate that the biochemical properties of the central serotonergic neurons in this culture system reflect reliably those of central serotonergic neurons in vivo. We suggest that measuring 5-HT and 5-HIAA contents in the primary cultured dissociated brainstem-cells with HPLC-EC is useful in the study of pharmacology as well as toxicolgy of the central serotonergic neurons.

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