• Title/Summary/Keyword: 신경세포막 특성

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원자간력현미경(AFM)을 활용한 SH-SY5Y Neuroblastoma 세포의 분화 전후 세포막의 Stiffness 변화 연구

  • Gwon, Sang-U;Yang, U-Cheol;Jeon, Gwang-Deok;Lee, Ji-Eun;Lee, Hui-Su;Jeong, Jin-Gwon;Hong, Hyeon-Min;Lee, Jeong-Min;Choe, Pu-Reum;Jang, Ju-Hui;Choe, Yun-Gyeong;Jeon, Song-Hui
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.599-599
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    • 2013
  • 최근의 원자간력현미경(AFM)은 생체물질을 대상으로 여러 구조적 형상뿐만 아니라 물리적 특성 측정이 가능하여 바이오분야에 다양이 활용되고 있다. 줄기세포의 신경세포로 분화 인지에 대한 연구와 관련하여 본 연구에서는 AFM의 한 기능인 Force-Distance curve 측정법을 활용하여 신경암세포주라 불리는 SH-SY5Y를 대상으로 분화 전과 후의 세포막의 stiffness 변화를 측정하였다. 세포막의 stiffness값은 시료표면과 맞닿은 AFM 탐침에 계속적으로 수직방향의 힘이 가해질 시 AFM 캔티레버의 구부러짐 정도로 측정된다. SH-SY5Y는 RA (retinoic acid) 처리에 의해 분화유도 되었으며, 생물학적 방법인 western blotting법을 통해 분화여부를 확인하였다. 측정영역은 AFM topography 이미지 상에서 roughness가 가장 낮은 분화 전과 후 SH-SY5Y의 핵 주변영역으로 선정하였다. 선정된 영역 내에 여러 부분의 분화 전후 세포막의 stiffness 값을 측정하여 통계화한 결과, 분화 전과 후 세포막의 stiffness 차이를 확인할 수 있었다. 분화 전 SH-SY5Y 세포막의 stiffness는 0.79445 N/m인 반면, 분화 후 SH-SY5Y 세포막의stiffness는 0.60324 N/m로 확인되었다. 이는 분화 전에 비하여 분화 후 SH-SY5Y 세포막의 stiffness가 약 24.07% 감소된 것으로 판단할 수 있다. 본 연구는 생물학적 복잡한 방법이 아닌 간단한 방법으로 세포의 stiffness의 변화 측정을 통한 세포의 분화를 판별할 수 있는 방법을 개발한 것으로 여러 줄기세포의 특정세포로 분화여부 판단에 활용할 수 있을 것으로 사료된다.

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CHANGES IN ELECTROPHYSIOLOGICAL PROPERTIES OF NEUROBIOTIN-LABELED PYRAMIDAL CELLS OF HIPPOCAMPUS RECORDED IN VIVO (마취된 흰쥐 해마신경세포에서 Neurobiotin 이온주입으로 인한 신경세포의 생리적 특성의 변화)

  • Lee, Hye-Sook;Lee, Maan-Gee-G.;Kim, Young-Jin;Choi, Byung-Ju
    • Journal of the korean academy of Pediatric Dentistry
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    • v.26 no.2
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    • pp.218-231
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    • 1999
  • Pyramidal cells in the hippocampal CA area were recorded from and filled with neurobiotin in anesthetized rats. The extent of their dendrites and the electropharmacological properties of membrane as well as the effect before and after neurobiotin injection were examined. Pyramidal cells had a high resting membrane potential, a low input resistance, and a large amplitude action potential. A afterhyperpolarization was followed a single action potential. Most pyramidal cells did not display a spontaneous firing. Pyramidal cell displayed weak inward rectification and anodal break excitation in response to negative current injection into the cell. Membrane properties of recorded neurons before and after neurobiotin injection with consecutive current injection were compared. Some properties were significantly increased after labelling(P>0.05); the duration and amplitude of sustained AHP, input resistance, and the number of action potentials for simultaneous intra- and extracellular stimulations. Neurobiotin-filled neurons showed pyramidal morphology. Cells were generally bipolar dendrite processes ramifying in stratum lacunosum-moleculare, radiatum, and oriens.

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ELECTROPHYSIOLOGICAL AND MORPHOLOGICAL CHARACTERISTICS OF FACIAL NUCLEUS IN RAT (흰쥐 안면신경핵 세포의 전기생리학적 및 형태학적 특성)

  • Choi, Byung-Ju;Cho, Jin-Hwa;Bae, Yong-Chul;Kim, Young-Jin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.27 no.3
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    • pp.400-409
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    • 2000
  • This study used in vivo intracellular and extracellular field potential recording to evaluate the intrinsic membrane properties and connection pattern within facial nucleus. 1. There were four subdivisions of medial, intermediate, lateral, and dorsolateral in facial nucleus. 2. Principal cells in the facial nucleus was recorded from and filled with neurobiotin in anesthetized rats. The extent of their dendrites and the characteristics of cell body were examined. 3. Principal cells had a large amplitude action potential and afterhyperpolarization was followed a single action potential. 4. The response from facial motonucleus to electrical stimulation of the facial nerve was mainly a monophasic wave, with a latency of 1 msec, which was assumed to reflect antidromic activation of facial motoneurons. In some of rats the response in addition showed late components at a latency of about 7-8 msec, but its amplitude was small. 5 Most of cells exhibited accommodation of spike discharge upon depolarization of membrane by 0.8 nA for 400 ms. Our results support the hypothesis that there normally are weak connections between different parts of the facial motonucleus to explain pathophysiology of hemifacial spasm and facial naive paralysis.

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A Study on the Electronic Potential Occurrance and Delevery Specific Property by ionic Density Transformation (이온농도 변화에 따른 뉴런의 전위발생과 전달특성에 관한 연구)

  • 김석환;류광렬;허창우
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 1999.11a
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    • pp.509-515
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    • 1999
  • 신체를 구성하는 여러 가지 요소 중에서 인간이 일상생활을 하는 데에는 신진대사를 일정하게 유지하기 위한 인체내의 여러 가지 전달물질들이 그 구성에 맞게 역할을 수행하여야 한다. 그러나, 만약 그 구성 비율이 바뀔 때에는 인체 내에 이상이 발생하게 된다. 그 대표적인 예로는 운동을 심하게 하였을 경우 갈증과 근육의 피로감이 몰려오게 되는데 이는 세포내의 이온농도의 변화에 의한 것이며 이때 물, 스포츠 이온 음료 등을 섭취한다. 순간적으로 그 증상이 회복되지는 않지만 신경계 내에서 회복하는데 에는 많은 시간이 소요된다. 생리학, 임상적으로 나트륨과 물은 신진대사에 밀접한 연관이 있다. 인체의 나트륨은 음식물의 섭취량과 나트륨의 생리작용사이의 균형에 좌우된다. 건강한 상태에서 나트륨의 생리적인 손실은 무시할 만 하며 나트륨 생리작용은 음식물의 섭취량과 밀접한 관련이 있다. 칼륨은 단지 전체 신경계 용질의 일부이기 때문에 이것은 세포의 체적과 세포 용질의 삼투현상을 결정하는 주요한 것이다. 더욱더 칼륨은 신진대사 과정의 많은 중요한 변수가 되며, 세포밖의 칼륨은 전체중에 매우 작은 일부이긴 하지만 신경근육 작용에 매우 큰 영향을 준다. 세포안과 밖의 칼륨의 농도 비율은 흥분성 조직내의 막 전위의 주요한 결정요소가 된다. 본연구에서는 세포막 설계 후 시뮬레이션을 통한 정상상태의 세포막 전위의 활동전위 전달 특성과 Na, K의 이온 농도를 변화시킬 경우의 세포막 전위의 활동저위 전달 특성곡선을 비교 관찰하여 운동 생리학적인 신경세포내의 전달특성 메카니즘을 이해, 분석하였다.서의 속도를 최소한으로 줄이면서 스케줄링에 의해 시간 문제를 해결하는 방법을 제시하도록 한다. 이는 기초과학 수준이 높은 북방권 국가들의 과학자들이 주로 활용되고 있다는 점에서도 잘 알 수 있으며 우리의 과학기술 약점을 보완하는 원천으로써 외국인 연구 인력이 대안이 되고 있음을 시사한다. 본 연구에서는 한국 연구 조직에서 일하는 외국인 연구자들의 동기 및 성과에 영향을 미치는 많은 요인들을 확인할 수 있었다. 상관관계, 분산분석, 회귀분석 등을 통해 활용 성과에 미치는 영향 요인들을 도출하였다. 설문 분석을 통하여 동기 및 성과 사이에는 강한 상관관계가 존재하는 것을 확인할 수 있었으며 이는 전통적인 동기 이론들과 부합한다. 대부분의 변수가 동기 및 성과에 동시에 영향을 미치는 것으로 조사되었으며 그중에서도 조직 협력 문화, 외국인 연구자의 의사소통 및 협력성, 외국인 연구자의 연구 능력 관련 변수들 및 연구 프로젝트의 기술수명주기, 외국인 연구자의 기존 기술지식의 흡수 등이 가장 중요한 변수로 나타났다. 이는 우리가 주로 중국 및 러시아 과학자들을 활용하여 상업화하는 외국인 연구인력 활용 패턴과도 일치하는 결과이다. 즉 우호적인 조직문화를 가지고 있는 연구 조직에서, 이미 과학기술 지식을 많이 가지고 있고 연구 능력도 높은 외국인 과학기술자를, 한국에서 기술이 태동 또는 성장하고 있는 연구 분야에서 활용하는 것이 가장 성과가 좋다는 사실을 확인시켜 주고 있다. 국내에서 최초로 수행된 본 연구는 외국인 연구 인력의 활용 성과가 매우 높으며, 우리의 과학기술혁신시스템을 보완하는

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The Role of G protein in the Activation of Phospholipase C from Bovine Brain (소의 뇌조직 Phospholipase C의 활성화에 미치는 G-단백질의 역할)

  • Kim, Jung-Hye;Lee, Dong-Jin;Byun, Yeung-Ju
    • Journal of Yeungnam Medical Science
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    • v.9 no.2
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    • pp.288-301
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    • 1992
  • The objective of the present study was to identify the characteristics of phospholipase C (PLC) isozymes purified from bovine brain and to investigate their interrelationship with G protein. The purified PLC isozymes ${\beta}$, ${\gamma}$ and ${\delta}$ were obtained and the characteristics of PLC activity on various concentrations of free $Ca^{2+}$ were observed. The activity of PLC was increased with increasing $Ca^{2+}$ concentration and the activity PLC ${\delta}$ was increased higher in the presence of phosphatidyl choline(PC) than in the abscence of PC. For vesicle formation as the structure of cell membrane, cholic acid and deoxycholic acid as detergent on phosphatidylinositol bisphosphate($PIP_2$) substrate containing PC were used, and then the activity of PLC isozymes were increased with increasing concentration of cholate, from 0.2% to 1% and were increased slightly in deoxycholate. In the $PIP_2$ containing phospholipid and glycolipid as brain extract, the activity of PLC isozymes were checked in 0.2%-1% cholic acid. The activities of PLC isoyzmes were continuously increased up to 1% cholic acid. The quantitation of PLC isozymes from several bovine organs by radioimmunoassay was made. Brain was the most sufficient organ in terms of amount of PLC ${\beta}$and ${\delta}$. A large amount of PLC ${\delta}$ was existed in adrenal gland. The binding capacity of GTPrS and G protein was observed and other observations of the binding effect of GTPrS-G protein and PLC monoclonal Ab-Protein A from tissue homogenate with PLC were made. From the observation the binding capacity was revealed the range of 0.11%-1.49%. The effects of each type of G protein on the percent activity of purified PLC isozymes were observed. From the observation, activities of isozymes were increased in $Go{\alpha}$ & Gmix, and the activities of PLC ${\beta}$ and ${\delta}$ were increased in $G{\beta}{\gamma}$ and $Gi{\alpha}$. Activities of PLC ${\beta}$ and ${\gamma}$ were decreased in $Gt{\alpha}$ but PLC ${\delta}$ increased.

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Ameliorating effect of the ethyl acetate fraction of Pteridium aquilinum on glucose-induced neuronal apoptosis (포도당으로 유도된 신경세포 손상에 대한 고사리 아세트산에틸 분획물의 개선 효과)

  • Park, Seon Kyeong;Guo, Tian Jiao;Kim, Jong Min;Kang, Jin Yong;Park, Sang Hyun;Kang, Jeong Eun;Kwon, Bong Seok;Lee, Chang Jun;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.49 no.4
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    • pp.430-437
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    • 2017
  • The protective effect of Pteridium aquilinum on high glucose-induced cytotoxicity was examined in vitro to investigate the relationship between diabetic condition and neuronal dysfunction. The ethyl acetate fraction of P. aquilinum (EFPA), with total phenolic content of 265.08 mg gallic acid equivalent/g, showed higher 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)/2,2-diphenyl-1-picrylhydrazyl radical scavenging activities and lipid peroxidation inhibitory effect than any other fraction. In addition, EFPA showed a significant reduction in the inhibitory effect on ${\alpha}$-glucosidase activity ($IC_{50}$ value=$205.26{\mu}g/mL$) compared to the acarbose positive control. The anti-oxidative effect in PC12 cells, protective effects on high glucose-induced oxidative stress in neuronal cells, and neurotoxicity were measured using 2',7'-dichlorofluorescin diacetate, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide, and lactate dehydrogenase assays, respectively. EFPA showed conspicuous inhibitory effect on cellular reactive oxygen species production and neuronal cell apoptosis. Finally, kaempferol-3-glucoside was identified as the main phenolic compound of EFPA using high performance liquid chromatography.

Independent Prognostic Value of the Fascin Expression in Patients with Esophageal Cancer (식도암에 있어서 Fascin의 발현과 예후와의 상관관계에 대한 연구)

  • Choi, Pill-Jo;Jeong, Sang-Seok;Bang, Jung-Heui;Cho, Kwang-Jo;Woo, Jong-Soo;Roh, Mee-Sook
    • Journal of Chest Surgery
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    • v.41 no.1
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    • pp.74-81
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    • 2008
  • Background: Fascin is an actin-bundling protein that induces membrane protrusions and it increases cell motility in various transformed cells. Esophageal cancer is one of the most lethal malignancies, and it exhibits extensive local invasion or frequent regional lymph node metastasis even after curative surgery. We investigate the expression of fascin by performing immunohistochemistry to evaluate the clinical characteristics and prognostic significance of its expression in esophageal cancer patients. Material and Method: Immunochemistry for fascin was performed on 76 tumor samples from 76 patients who underwent esophageal cancer operations. The expression levels of fascin in the 76 esophageal cancer tissues were compared with those in the corresponding normal esophageal epithelium. The fascin-positive samples were defined as those showing more than 75% of fascin-positive cells. Result: Overall, a fascin positive expression was detected in 39 (51.3%) out of the total 76 cases. The tumors with positive fascin expression tended to more frequently show a higher stage (p=0.030), and a higher T-factor (p=0.031). The prognosis of the fascin negative group was significantly better than that of the fascin positive group (p=0.004). Multivariate analysis revealed that lymphovascular invasion and the fascin expression were independent prognostic factors. Conclusion: Fascin was expressed in 513% of the esophageal cancer tissues and a positive expression of fascin was associated with more advanced tumor progression and recurrence. Our study suggests that the fascin expression may be an independent prognostic factor for an unfavorable clinical course few those patients suffering with esophageal cancer.

The Relationship between Intracellular Protein Kinase C Concentration and Invasiveness in U-87 Malignant Glioma Cells (교모세포종 세포주 U-87에서 세포내 PKC 농도와 종양침습성과의 상관 관계)

  • Ji, Cheol;Cho, Kyung-Keun;Lee, Kyung Jin;Park, Sung Chan;Cho, Jung Ki;Kang, Joon Ki;Choi, Chang Rak
    • Journal of Korean Neurosurgical Society
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    • v.30 no.3
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    • pp.263-271
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    • 2001
  • Objective : Glioblastomas, the most common type of primary brain tumors, are highly invasive and cause massive tissue destruction at both the tumor invading edges and in areas that are not in direct contact with glioma cells. As a result, patients with high-grade gliomas are faced with a poor prognosis. Such grim statistics emphasize the need to better understand the mechanisms that underlie glioma invasion, as these may lead to the identification of novel targets in the therapy of high grade gliomas. Protein kinase C(PKC) is a family of serine/threonine kinases and an important signal transduction enzyme that conveys signals generated by ligand-receptor interaction at the cell surface to the nucleus. PKC appears to be critical in regulating many aspects of glioma biology. The purpose of this study was to assess accurately the role of PKC in the invasion regulation of human gliomas based on hypothesis that protein kinase C(PKC) is functional in the process of glial tumor cell invasion. Method : To test this hypothesis, U-87 malignant glioma cell line intracellular PKC levels were up and down regulated and their invasiveness was tested. Intracellular PKC level was characterized using PKC activity assays. Invasion assays including barrier migration and spheroid confrontation were used to study the relationship between PKC concentration and invasiveness. Result : The cell line which were treated by PKC inhibitor tamoxifen and hypericin exhibited decreased PKC activity and decreased invasive abilities dose dependently both in matrigel invasion assay and tumor spheroid fetal rat brain aggregates(FRBA) confrontation assay. However, the cell line that was treated by PKC activator 12-O-tetradecanylphorbol-13acetate(TPA) did not exhibit increases in either PKC activity or invasive ability. Conclusion : These studies suggest that PKC may be a useful molecular target for the chemotherapy of glioblastoma and other malignancies and that a therapeutic approach based on the ability of PKC inhibitors may be helpful in preventing invasion.

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