• Title/Summary/Keyword: culture conditions

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The optimal conditions to improve retrovirus-mediated transduction efficiency to NIH 3T3 cells (레트로바이러스(retrovirus)의 NIH 3T3 세포로의 유전자 전달효율을 증가시키기 위한 적절한 조건들)

  • Lee, Jun Ah;Lee, Kang-Min;Lee, Hyun Jae;Lee, Yun Jeong;Kim, Dong Ho;Lim, Jung Sub;Park, Kyung-Duk
    • Clinical and Experimental Pediatrics
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    • v.50 no.10
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    • pp.1011-1017
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    • 2007
  • Purpose : We tried to assess the optimal conditions to improve low transduction efficiency and their effect on target cells. Methods : Cultured NIH 3T3 cells were incubated with retroviral vectors bearing an enhanced green fluorescent protein (eGFP) gene. We varied the ratio of viral vectors to target cells (1:1-1:8) and the number of transfections (${\times}1$, ${\times}2$), and compared transduction efficiencies. Also, the effects of polybrene on transduction efficiency and viability of target cells were assessed. Transduction of the eGFP gene was evaluated by observing NIH 3T3 cells under a fluorescence microscope and efficiencies were measured by the percentage of eGFP positive cells using FACscan. Results : As the ratio of retroviral vectors to target cells increased, transduction efficiency was greatly improved, from 7% (1:1) to 38% (1:4). However, transduction efficiency did not increase any more when the ratio increased from 1:4 to 1:8. Cells transfected twice showed higher transduction efficiencies than cells transfected once, at a ratio of 1:8. The eGFP gene transduced to NIH 3T3 cells sustained its expression during repeated passages. However, after the third passage (day 9), the percentage of eGFP positive cells began to decline. The degree of this decline in eGFP expression was lower in cells transfected twice than in cells transfected once (P<0.05). The addition of polybrene did not have any toxic effect on NIH 3T3 cells and greatly increased transduction efficiency (P=0.007). In addition to vector component, transduction efficiency was very sensitive to culture confluence. Cells cultured and transfected in 24-well plate showed higher transduction efficiency, although cells cultured in 6- well plate proliferated more (P=0.024). Conclusion : Our data could be used as a basis for retrovirus-based gene therapy. Further study will follow using human cells as target cells.

Growth responses of kelp species Ecklonia cava to different temperatures and nitrogen sources (온도와 질소원 종류에 따른 대형갈조류 감태(Ecklonia cava)의 생장)

  • Choi, Sun Kyeong;Kang, Yun Hee;Park, Sang Rul
    • Korean Journal of Environmental Biology
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    • v.38 no.3
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    • pp.404-415
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    • 2020
  • We investigated the seasonal variations in growth and physiological responses of the kelp species Ecklonia cava to different nitrogen sources to establish indoor culture conditions for mass production. Ecklonia cava was cultivated for 10 days in 16 combinations of seawater temperatures (15, 17, 21, and 25℃) and different nitrogen sources (control; NH-NH4+ 100 μM; NO-NO3- 100 μM; and NHNO-NH4+ 50 μM+NO3- 50 μM). The growth and growth rate of the blade were affected by temperature. The mean fresh weight and area-based daily growth rate were the highest (5.8±0.5 and 6.6±0.5% day-1, respectively) at 15℃ and the lowest (2.2±0.2 and 3.0±0.3% day-1, respectively) at 25℃. The daily growth rate was the highest in the NH and NO treatments and lowest in the control. The nitrate reductase activity of E. cava varied with water temperature (season). The highest activity was in the control (1.32±0.10 μmol NO2- g-1 dry weight h-1) and the lowest was in the NH treatment (0.25±0.02 μmol NO2- g-1 dry weight h-1). The photosynthetic pigment concentrations reached a maximum value in the NHNO treatment and a minimum value in the control. These results showed that water temperature played an important role in the cultivation of E. cava and that a single supply of NH4+ or NO3- may induce the accelerated growth of E. cava. The growth and physiological responses of E. cava to different nitrogen sources during each season provide valuable information for determining the optimal nitrogen source in E. cava cultivation under indoor conditions.

Trend and Further Research of Rice Quality Evaluation (쌀의 품질평가 현황과 금후 연구방향)

  • Son, Jong-Rok;Kim, Jae-Hyun;Lee, Jung-Il;Youn, Young-Hwan;Kim, Jae-Kyu;Hwang, Hung-Goo;Moon, Hun-Pal
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.47
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    • pp.33-54
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    • 2002
  • Rice quality is much dependent on the pre-and post harvest management. There are many parameters which influence rice or cooked rice qualitys such as cultivars, climate, soil, harvest time, drying, milling, storage, safety, nutritive value, taste, marketing, eating, cooking conditions, and each nations' food culture. Thus, vice evaluation might not be carried out by only some parameters. Physicochemical evaluation of rice deals with amy-lose content, gelatinizing property, and its relation with taste. The amylose content of good vice in Korea is defined at 17 to 20%. Other parameters considered are as follows; ratio of protein body-1 per total protein amount in relation to taste, and oleic/linoleic acid ratio in relation to storage safety. The rice higher Mg/K ratio is considered as high quality. The optimum value is over 1.5 to 1.6. It was reported that the contents of oligosaccharide, glutamic acid or its derivatives and its proportionalities have high corelation with the taste of rice. Major aromatic compounds in rice have been known as hexanal, acetone, pentanal, butanal, octanal, and heptanal. Recently, it was found that muco-polysaccharides are solubilized during cooking. Cooked rice surface is coated by the muco-polysaccharide. The muco-polysaccharide aye contributing to the consistency and collecting free amino acids and vitamins. Thus, these parameters might be regarded as important items for quality and taste evaluation of rice. Ingredients of rice related with the taste are not confined to the total rice grain. In the internal kernel, starch is main component but nitrogen and mineral compounds are localized at the external kernel. The ingredients related with taste are contained in 91 to 86% part of the outside kernel. For safety that is considered an important evaluation item of rice quality, each residual tolerance limit for agricultural chemicals must be adopted in our country. During drying, rice quality can decline by the reasons of high drying temperature, overdrying, and rapid drying. These result in cracked grain or decolored kernel. Intrinsic enzymes react partially during the rice storage. Because of these enzymes, starch, lipid, or protein can be slowly degraded, resulting in the decline of appearance quality, occurrence of aging aroma, and increased hardness of cooked rice. Milling conditions concerned with quality are paddy quality, milling method, and milling machines. To produce high quality rice, head rice must contain over three fourths of the normal rice kernels, and broken, damaged, colored, and immature kernels must be eliminated. In addition to milling equipment, color sorter and length grader must be installed for the production of such rice. Head rice was examined using the 45 brand rices circulating in Korea, Japan, America, Australia, and China. It was found that the head rice rate of brand rice in our country was approximately 57.4% and 80-86% in foreign countries. In order to develop a rice quality evaluation system, evaluation of technics must be further developed : more detailed measure of qualities, search for taste-related components, creation and grade classification of quality evaluation factors at each management stage of treatment after harvest, evaluation of rice as food material as well as for rice cooking, and method development for simple evaluation and establishment of equation for palatability. On policy concerns, the following must be conducted : development of price discrimination in conformity to rice cultivar and grade under the basis of quality evaluation method, fixation of head rice branding, and introduction of low temperature circulation.

New horizon of geographical method (인문지리학 방법론의 새로운 지평)

  • ;Choi, Byung-Doo
    • Journal of the Korean Geographical Society
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    • v.38
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    • pp.15-36
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    • 1988
  • In this paper, I consider the development of methods in contemporary human geography in terms of a dialectical relation of action and structure, and try to draw a new horizon of method toward which geographical research and spatial theory would develop. The positivist geography which was dominent during 1960s has been faced both with serious internal reflections and strong external criticisms in the 1970s. The internal reflections that pointed out its ignorance of spatial behavior of decision-makers and its simplication of complex spatial relations have developed behavioural geography and systems-theoretical approach. Yet this kinds of alternatives have still standed on the positivist, geography, even though they have seemed to be more real and complicate than the previous one, The external criticisms that have argued against the positivist method as phenomenalism and instrumentalism suggest some alternatives: humanistic geography which emphasizes intention and action of human subject and meaning-understanding, and structuralist geography which stresses on social structure as a totality which would produce spatial phenomena, and a theoretical formulation. Human geography today can be characterized by a strain and conflict between these methods, and hence rezuires a synthetic integration between them. Philosophy and social theory in general are in the same in which theories of action and structural analysis have been complementary or conflict with each other. Human geography has fallen into a further problematic with the introduction of a method based on so-called political ecnomy. This method has been suggested not merely as analternative to the positivist geography, but also as a theoretical foundation for critical analysis of space. The political economy of space with has analyzed the capitalist space and tried to theorize its transformation may be seen either as following humanistic(or Hegelian) Marxism, such as represented in Lefebvre's work, or as following structuralist Marxism, such as developed in Castelles's or Harvey's work. The spatial theory following humanistic Marxism has argued for a dialectic relation between 'the spatial' and 'the social', and given more attention to practicing human agents than to explaining social structures. on the contray, that based on structuralist Marxism has argued for social structures producing spatial phenomena, and focused on theorising the totality of structures, Even though these two perspectives tend more recently to be convergent in a way that structuralist-Marxist. geographers relate the domain of economic and political structures with that of action in their studies of urban culture and experience under capitalism, the political ecnomy of space needs an integrated method with which one can overcome difficulties of orthhodox Marxism. Some novel works in philosophy and social theory have been developed since the end of 1970s which have oriented towards an integrated method relating a series of concepts of action and structure, and reconstructing historical materialism. They include Giddens's theory of structuration, foucault's geneological analysis of power-knowledge, and Habermas's theory of communicative action. Ther are, of course, some fundamental differences between these works. Giddens develops a theory which relates explicitly the domain of action and that of structure in terms of what he calls the 'duality of structure', and wants to bring time-space relations into the core of social theory. Foucault writes a history in which strategically intentional but nonsubjective power relations have emerged and operated by virtue of multiple forms of constrainst wihthin specific spaces, while refusing to elaborate any theory which would underlie a political rationalization. Habermas analyzes how the Western rationalization of ecnomic and political systems has colonized the lifeworld in which we communicate each other, and wants to formulate a new normative foundation for critical theory of society which highlights communicatie reason (without any consideration of spatial concepts). On the basis of the above consideration, this paper draws a new norizon of method in human geography and spatial theory, some essential ideas of which can be summarized as follows: (1) the concept of space especially in terms of its relation to sociery. Space is not an ontological entity whch is independent of society and has its own laws of constitution and transformation, but it can be produced and reproduced only by virtue of its relation to society. Yet space is not merlely a material product of society, but also a place and medium in and through which socety can be maintained or transformed.(2) the constitution of space in terms of the relation between action and structure. Spatial actors who are always knowledgeable under conditions of socio-spatial structure produce and reproduce their context of action, that is, structure; and spatial structures as results of human action enable as well as constrain it. Spatial actions can be distinguished between instrumental-strategicaction oriented to success and communicative action oriented to understanding, which (re)produce respectively two different spheres of spatial structure in different ways: the material structure of economic and political systems-space in an unknowledged and unitended way, and the symbolic structure of social and cultural life-space in an acknowledged and intended way. (3) the capitalist space in terms of its rationalization. The ideal development of space would balance the rationalizations of system space and life-space in a way that system space providers material conditions for the maintainance of the life-space, and the life-space for its further development. But the development of capitalist space in reality is paradoxical and hence crisis-ridden. The economic and poltical system-space, propelled with the steering media like money, and power, has outstriped the significance of communicative action, and colonized the life-space. That is, we no longer live in a space mediated communicative action, but one created for and by money and power. But no matter how seriously our everyday life-space has been monetalrized and bureaucratised, here lies nevertheless the practical potential which would rehabilitate the meaning of space, the meaning of our life on the Earth.

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Effect of Crude Polysaccharide Isolated from Mageoli Mash under Different Fermentation Conditions on Immune-Stimulating Activity (발효 조건이 막걸리 술덧 유래 다당의 면역자극 활성에 미치는 효과)

  • Rhee, Young Kyoung;Nam, So-Hyun;Kim, Hye-Ryun;Cho, Chang-Won;Lee, Young-Chul;Kim, Young-Chan;Hong, Hee-Do
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.12
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    • pp.1835-1842
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    • 2014
  • Various microorganisms are involved in the fermentation of Mageoli, Korean traditional liquor. Among them, the most predominant microorganism is yeast, including Saccharomyces cerevisae. This study investigated the effects of crude polysaccharides separated from Mageoli mash (CP-M) on activation of peritoneal macrophages in cell culture medium. Four types of yeasts, S. cerevisae 89-1-1, 98-2, 268-3, and 113-4, were used. When peritoneal macrophages were treated with $10{\mu}g/mL$ of CP-M fermented with S. cerevisae 113-4, the concentration of nitric oxide (NO) in the medium was highest ($33.3{\mu}M$). The concentrations of IL-6 and -12 were also highest at 116.3 pg/mL and 59.8 pg/mL, respectively. In the case of CP-M cultured at 15, 20, and $30^{\circ}C$ after inoculation with S. cerevisae 113-4 and fermented at different temperatures, production of NO and IL-6 by peritoneal macrophages did not change compared to the control. For CP-M obtained after fermentation at $25^{\circ}C$, however, NO concentration increased 2.7~3.3 fold and IL-6 concentration by 5.7 fold compared to the control. Furthermore, the effect of fermentation period on expression of CP-M functionality was examined. NO production by CP-M of the 5th day mash was highest, increasing 2.2 fold compared to 0-day CP-M. However, there were no significant differences in concentration of IL-6 or -12 according to different fermentation periods, although CP-M showed a large decrease after 10 days. The results show that fermentation conditions with the highest activity were observed in CP-M inoculated with S. cerevisae 113-4 and fermented at $25^{\circ}C$ for 5 days. CP-M consisted of 78.6% neutral sugar, 11.6% acidic polysaccharide, and 9.8% protein. In particular, the sugar composition of neutral sugar consisted of mannose (47.8%), glucose (29.6%), and galactose (12.7%). Based on the results, CP-M is assumed to be an extracellular polysaccharide originating in yeast with high mannose content.

Biological Characteristics and Growth of the Korean Freshwater Rotifer, Brachionus calyciflorus at Various Temperatures (한국 담수산 윤층 Brachionus calyciflorus의 생물학적 특징과 온도별 성장)

  • 강언종;이배익;김응오
    • Journal of Aquaculture
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    • v.10 no.4
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    • pp.449-456
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    • 1997
  • This study was concuted to determine the optimal conditions for raising the freshwater rotifer, Brachinus calyciflorus. The authors presented some biological informatin obtained from incubation experiment under the various controlled temperatures. Lorica size of the rotifer was divided into two groups : the length and the width for the S-type was $141.0\pm16.7\mu m$($110.1-182.5\;\mu m, n=44$)and $107.0\pm20.3\mu m\;(75.3-152.3\mu m, n=44)$, and those for the L-type was $262.8\pm15.2\mu m\;(234.4-288.6\mu m,\;n=20)\;and\;182.6\pm13.4\mu m (159.8-207.0\mu m,\;n=20$), respectively. The number of eggs being attached on the female varied from 1 to 11 at various culture conditions. Egg type was divided into two groups, large and small. Large and small egg was measured in its major axis as 85a.7-107.8$\mu$m and 55.1-65.2$\mu$m for S-type, and 104.9-121.8 $\mu$m and 62.8-89.1$\mu$m for L-type respectively. The maximum density was reached at 4th day after incubation. The density was 583.9 rotifers/$m\ell$ for $25^{\circ}C$-experimental. group and 421.3 rotifers/$m\ell$ for $22^{\circ}C$-experimental. group respectively. In the case of $28^{\circ}C$-experimental. group, it suddenly decreased into 4.7 rotifers/$m\ell$ at 1st day after incubations and did not recover to its initial density. The maximum rate of increase of populatin per day was reached 0.802 for $22^{\circ}C$-experimental. group at day 2 and fluctuated thereafter. For $25^{\circ}C$-experimental. group it increased to 0.964 at day 3 of incubation and then declined. And the egg ratio of female was reached the maximum of 0.614 for 22$^{\circ}C$- at 3rd day and 0.772 for $25^{\circ}C$-experimental. group at 4th day of incubation.

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Studies on the Manufacture of Concentrated Feed by the Use of Farm Product Waste Materials (농산물(農産物) 폐물(廢物)을 이용(利用)한 농후사료(濃厚飼料) 제조(製造)에 관(關)한 연구(硏究))

  • Kim, Sam-Soon;Lee, Ji-Yul;Park, Sung-Oh;Kim, Ki-Joo
    • The Korean Journal of Mycology
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    • v.1 no.2
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    • pp.15-23
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    • 1973
  • Mold producing cellulase were isolated from rotten woods, and identified as the three species: Aspergillus niger van Tieghem, Aspergillus schiemanni Thom and Trichoderma viride Pers. In this paper, culture conditions in the media and characteristics of these strains were investigated. Using these strains, we have conducted a research concerning the utilization of farm product waste materia's. 1. Optimum conditions for the cellulase formation were as follows. KM 10-1; pH 5.2-5.5, $35^{\circ}C$, incubation time 6 days. OL 11-1; pH 5.5, $30-35^{\circ}C$, incubation time 6 days. SH 9-2; pH 5.5, $30^{\circ}C$, incubatoin time 6 days. 2. Their cellulase activities in their optimum condition were as follows: KM 10-1; CMC-LP 78.5% CMC-SP 4.0 glucose mg/gm of the cultures/min. OL 11-1; CMC-LP 89.9%, CMC-SP 4.9 glucose mg/gm of the cultures/min. SH 9-2; CMC-I.P 77.4%, CMC-SP 3.9 glucose mg/gm of the cultures/min. 3. Hydrolysis of animal feed containing a large quantity (23-30%) of cellulose by means of the crude enzyme in the selected strains resolved 30% of the cellulose contained in the animal feed.

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Purification of Human HtrA1 Expressed in E. coli and Characterization of Its Serine Protease Activity (E. coli에서 발현된 human HtrA1 단백질의 정제와 HtrA1의 serine protease 활성 조건에 관한 연구)

  • Kim, Kyung-Hee;Kim, Sang-Soo;Kim, Goo-Young;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.16 no.7 s.80
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    • pp.1133-1140
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    • 2006
  • Human HtrA1 (High temperature requirement protein A1) is a homologue of the E. coli periplasmic serine protease HtrA. A recent study has demonstrated that HtrA1 is a serine protease involved in processing of insulin like growth factor binding protein (ICFBP), indicating that it serves as an important regulator of IGF activity. Additionally, several lines of evidence suggest a striking correlation between proteolytic activity of HtrA1 serine protease and the pathogenesis of several diseases; however, physiological roles of HtrA1 remain to be elucidated. We used the pGEX bacterial expression system to develop a simple and rapid method for purifying HtrA1, and the recombinant HtrA1 protein was utilized to investigate the optimal conditions in executing its proteolytic activity. The proteolytically active HtrA1 was purified to approximately 85% purity, although the yield of the recombinant HtrA1 protein was slightly low $460{\mu}g$ for 1 liter E. coli culture). Using in vitro endoproteolytic cleavage assay, we identified that the HtrA1 serine protease activity was dependent on the enzyme concentration and the incubation time and that the best reaction temperature was $42^{\circ}C$ instead of $37^{\circ}C$. We arbitrary defined one unit of proteolytic activity of the HtrA1 serine protease as 200nM of HtrA1 that cleaves half of $5{\mu}M\;of\;{\beta}-casein$ during 3 hr incubation at $37^{\circ}C$. Our study provides a method for generating useful reagents to investigate the molecular mechanisms by which HtrA1 serine protease activity contributes in regulating its physiological function and to identify natural substrates of HtrA1.

Studies on the Improvement and Utilization of Pasture in the Foerst II. The effect of shde on the intake palptability and grazing behavior (임간초지의 개량 및 이용에 관한 연구 II. 채식기호성 및 방목습성에 미치는 비음의 영향)

  • 이인덕;윤익석;이조윤;신용국
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.5 no.3
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    • pp.207-211
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    • 1985
  • This study was conducted to determine the effects of shade (full day light; 100, 80, 60, 40 and 20%) and fertilization levels (high; $N\;30kg-P_2O_5\;35kg-K_2O\;30kg$, moderate; $N\;20kg-P_2O_5\;25kg-K_2O\;20kg$, and low; $N\;10kg-P_2O_5\;15kg-K_2O\;10kg$) of forest pasture on intake palatability of Korean native goat, and the effects of shade under the conditions of 50% shaded forest pasture and full day light pastures on grazing behavior. This experiment was done at the experimental field of Chungnam National University, in Taejon, from 1983 to 1984. The results obtained are summarized as follows: 1. The dry matter intake, R.I.I. and Touch number of Korean native goat were affected by the shade degrees and fertilization levels, generally the increase of the fertilization level and high shading degree increased the NO_3-N$ contents in the plants, but decreased the soluble carbohydrate contents. It means that the decrease of the utilization of the shade culture grasses is due to the decreased intake palatability of forest pastures, particularly the conditions high fertilization level and above 60% of shading level seemed to have a nitrate stress. 2. Grazing time of forest pastures by Korean native goat tended to be spend more time than full day light pastures, also tendency of the major grazing activity tended to be graze continuously without difference of variation of grazing time within a day. Considering of the characteristies os shade, the intensive utilization of forest pastures seemed to be most effective during the high temperature season in August.

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A Study on the Effects of Sirtuin 1 on Dendritic Outgrowth and Spine Formation and Mechanism in Neuronal Cells (신경세포에서 sirtuin 1이 수상돌기 성장과 가시형성에 미치는 영향 및 기전에 관한 연구)

  • Seo, Mi Kyoung;Kim, Hye Kyeong;Baek, Song Young;Lee, Jung Goo;Urm, Sang-Hwa;Park, Sung Woo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.31 no.9
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    • pp.806-817
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    • 2021
  • Increasing evidence suggests that depression is associated with impairments in neural plasticity. Sirtuin 1 plays an important role in neural plasticity, and the activation of mechanistic target of rapamycin complex 1 (mTORC1) signaling is known to improve neural plasticity. In this study, we aimed to determine whether sirtuin 1 affects dendrite outgrowth and spine formation through mTORC1 signaling. Resveratrol (sirtuin 1 activator; 1 and 10 μM) and sirtinol (sirtuin 1 inhibitor; 1 and 10 μM) were treated in primary cortical culture with and without dexamethasone (500 μM). Levels of sirtuin 1, phospho-extracellular signal regulated protein kinase 1/2 (ERK1/2), phospho-mTORC1, and phospho-p70 ribosomal protein S6 kinase (p70S6K) were evaluated using Western blot analysis. Dendritic outgrowth and spine density were assessed using immunostaining. Resveratrol significantly increased levels of sirtuin 1 expression and phosphorylation of ERK1/2 (a downstream target of sirtuin 1), mTORC1, and p70S6K (a downstream target of mTORC1) in a concentration-dependent manner under dexamethasone conditions. Resveratrol also significantly increased dendritic outgrowth and spine density. Conversely, sirtinol significantly decreased levels of sirtuin 1 expression and phosphorylation of ERK1/2, mTORC1, and p70S6K in a concentration-dependent manner under normal conditions. Moreover, sirtinol significantly decreased dendritic outgrowth and spine density. Consistent with the results of sirtinol, sirtuin 1 knockdown using sirtuin 1 siRNA transfection significantly decreased dendritic outgrowth and spine density as well as phosphorylation levels of ERK1/2 and mTORC1. These data suggest that sirtuin 1 enhances dendritic outgrowth and spine density by activating mTORC1 signaling.