• 제목/요약/키워드: brain slice

검색결과 139건 처리시간 0.027초

Oxygen/Glucose Deprivation and Reperfusion Cause Modifications of Postsynaptic Morphology and Activity in the CA3 Area of Organotypic Hippocampal Slice Cultures

  • Jung, Yeon Joo;Suh, Eun Cheng;Lee, Kyung Eun
    • The Korean Journal of Physiology and Pharmacology
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    • 제16권6호
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    • pp.423-429
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    • 2012
  • Brain ischemia leads to overstimulation of N-methyl-D-aspartate (NMDA) receptors, referred as excitotoxicity, which mediates neuronal cell death. However, less attention has been paid to changes in synaptic activity and morphology that could have an important impact on cell function and survival following ischemic insult. In this study, we investigated the effects of reperfusion after oxygen/glucose deprivation (OGD) not only upon neuronal cell death, but also on ultrastructural and biochemical characteristics of postsynaptic density (PSD) protein, in the stratum lucidum of the CA3 area in organotypic hippocampal slice cultures. After OGD/reperfusion, neurons were found to be damaged; the organelles such as mitochondria, endoplasmic reticulum, dendrites, and synaptic terminals were swollen; and the PSD became thicker and irregular. Ethanolic phosphotungstic acid staining showed that the density of PSD was significantly decreased, and the thickness and length of the PSD were significantly increased in the OGD/reperfusion group compared to the control. The levels of PSD proteins, including PSD-95, NMDA receptor 1, NMDA receptor 2B, and calcium/calmodulin-dependent protein kinase II, were significantly decreased following OGD/reperfusion. These results suggest that OGD/reperfusion induces significant modifications to PSDs in the CA3 area of organotypic hippocampal slice cultures, both morphologically and biochemically, and this may contribute to neuronal cell death and synaptic dysfunction after OGD/reperfusion.

해마절편의 허혈성 $K^+$ 축적에 대한 $K^+$채널 조절 약물의 작용 (Effects of $K^+$ Channel Modulators on Extracellular $K^+$ Accumulation during Ischemia in the Rat Hippocampal Slice)

  • 최진규;전보권;류판동
    • The Korean Journal of Physiology and Pharmacology
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    • 제1권6호
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    • pp.681-690
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    • 1997
  • Loss of synaptic transmission and accumulation of extracellular $K^+([K^+]_O)$ are the key features in ischemic brain damage. Here, we examined the effects of several $K^+$channel modulators on the early ischemic changes in population spike (PS) and $[K^+]_o$ in the CA1 pyramidal layer of the rat hippocampal slice using electrophysiological techniques. After onset of anoxic aglycemia (AA), orthodromic field potentials decreased and disappeared in $3.3{\pm}0.22\;min$ $(mean{\pm}SEM,\;n=40)$. The hypoxic injury potential (HIP), a transient recovery of PS appeared at $6.0{\pm}0.25\;min$ (n=40) in most slices during AA and lasted for $3.3{\pm}0.43\;min$. $[K^+]_o$ increased initially at a rate of 0.43 mM/min (Phase 1) and later at a much faster rate (12.45 mM/min, Phase 2). The beginning of Phase 2 was invariably coincided with the disappearance of HIP. Among $K^+$ channel modulators tested such as 4-aminopyridine (0.03, 0.3 mM), tetraethylammonium (0.1 mM), NS1619 $(0.3{\sim}10\;{\mu}M)$, niflumic acid (0.1 mM), glibenclamide $(40\;{\mu}M)$, tolbutamide $(300\;{\mu}M)$ and pinacidil $(100\;{\mu}M)$, only 4-aminopyridine (0.3 mM) induced slight increase of $[K^+]_o$ during Phase 1. However, none of the above agents modulated the pattern of Phase 2 in $[K^+]_o$ in response to AA. Taken together, the experimental data suggest that 4-aminopyridine-sensitive $K^+$channels, large conductance $Ca^{2+}-activated$ $K^+$ channels and ATP-sensitive $K^+$ channels may not be the major contributors to the sudden increase of $[K^+]_o$ during the early stage of brain ischemia, suggesting the presence of other routes of $K^+$ efflux during brain ischemia.

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Fluoxetine Modulates Corticostriatal Synaptic Transmission through Postsynaptic Mechanism

  • Cho, Hyeong-Seok;Choi, Se-Joon;Kim, Ki-Jung;Lee, Hyun-Ho;Cho, Young-Jin;Kim, Seong-Yun;Sung, Ki-Wug
    • The Korean Journal of Physiology and Pharmacology
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    • 제10권1호
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    • pp.31-38
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    • 2006
  • Fluoxetine, widely used for the treatment of depression, is known to be a selective serotonin reuptake inhibitor (SSRI), however, there are also reports that fluoxetine has direct effects on several receptors. Employing whole-cell patch clamp techniques in rat brain slice, we studied the effects of fluoxetine on corticostriatal synaptic transmission by measuring the change in spontaneous excitatory postsynaptic currents (sEPSC). Acute treatment of rat brain slice with fluoxetine ($10{\mu}M$) significantly decreased the amplitude of sEPSC ($8.1{\pm}3.3$%, n=7), but did not alter its frequency ($99.1{\pm}4.7$%, n=7). Serotonin ($10{\mu}M$) also significantly decreased the amplitude ($81.2{\pm}3.9$%, n=4) of sEPSC, but did not affect its frequency ($105.8{\pm}8.0$, n=4). The effect of fluoxetine was found to have the same trend as that of serotonin. We also found that the inhibitory effect of fluoxetine on sEPSC amplitude ($93.0{\pm}1.9$%, n=8) was significantly blocked, but not serotonin ($84.3{\pm}1.6$%, n=4), when the brain slice was incubated with p-chloroamphetamine ($10{\mu}M$), which depletes serotonin from the axon terminals and blocks its reuptake. These results suggest that fluoxetine inhibits corticostriatal synaptic transmission through postsynaptic, and that these effects are exerted through both serotonin dependent and independent mechanism.

MR영상의 3차원 가시화 및 분석을 위한 뇌영역의 자동 분할 (Automatic Brain Segmentation for 3D Visualization and Analysis of MR Image Sets)

  • 김태우
    • 한국정보처리학회논문지
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    • 제7권2호
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    • pp.542-551
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    • 2000
  • 본 논문에서는 MR 영상의 3차원 가시화 및 분석을 위한 단일 채널 MR 영상의 자동 뇌영역 분할 방법을 제안한다. 이 방법은 4단계의 2차원 및 3차원 처리에 의하여 뇌윤곽을 찾아낸다. 1,2단계에서는 곡선 적합을 이용한 자동 문턱치화에 의하여 머리마스크와 초기 뇌마스크를 생성한다. 3단계에서 입방보간으로 초기 뇌마스크의 3차원 볼륨을 생성하여 형태학적 연산, 연결부위 레이블링에 의하여 중기 뇌마스크를 생성한다. 최종적으로 곡선 적합에 의한 자동 문턱치화를 이용하여 뇌마스크를 정련한다. 제안한 알고리즘은 영상의 슬라이스 방향을 고려할 필요가 없고 영상이 뇌 전체를 포함하지 않아도 되며, T1, T2, PD, SPGR등 다양한 종류의 MR 영상의 자동적인 뇌영역의 분할에 유용하다. 실험에서 20세트 MR 영상에 대하여 수동분할을 기준으로 0.97 이상의 유지도를 보였다.

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개의 발에 염증 유발 후 전침이 통증역치 및 뇌줄기 큰솔기핵의 5-Hydroxytryptamine 합성에 미치는 효과 (The Effect of Electroacupuncture on Pain Threshold and 5.Hydroxytryptamine Synthesis Induced by Nociception in the Raphe Magnus Nucleus of Dog Brain Stem)

  • 조미숙
    • The Journal of Korean Physical Therapy
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    • 제23권1호
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    • pp.59-66
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    • 2011
  • Purpose: The purpose of this study was to investigate the effect of electroacupuncture on pain threshold and 5-HT synthesis in raphe magnus nucleus of dog brain stem following nociception induced by 1% carageenan. Method: Twenty male dogs were randomly divided into two groups, a control group and an electroacupuncture group with the following parameters: ST36, LI11 and SP9 with 120Hz and 0.5mA. The dogs were sacrificed at 30, 60, 90, 120, 150 and 180 min after nociception using 1% carageenan injection into the plantar side of the dog's left leg. The pain threshold was recorded by an Analgesia-meter. After making brain slice sections, the tissues were immunostained with 5-HT antisera (1:2,500). Results: The pain threshold for the electroacupuncture group was higher than for the control group. The number of 5-HT immunoreactive neurons in the electroacupuncture group was greater than that of the control group. Differences between the two groups regarding the number of 5-HT immunoreactive neurons at 30 min and 60 min (p<0.05), at 120 min and 150 min (p<0.01), and at 8 days (p<0.05) were significant. Conclusion: The increases in pain threshold and number of 5-HT immunoreactive neurons in the raphe magnus nucleus of dog brain stem showed that electroacupuncture can attenuate pain transduction through the descending tract.

뇌전이 환자의 조영 증강 후 지연 FLAIR 영상의 유용성 (Patients with brain metastases the usefulness of contrast-enhanced FLAIR images after delay)

  • 변재후;박명환;이진완
    • 대한디지털의료영상학회논문지
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    • 제16권1호
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    • pp.13-19
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    • 2014
  • Purpose: FLAIR image is beneficial for the diagnosis of various bran diseases including ischemic CVS, brain tumors and infections. However the border between the legion of brain metastasis and surrounding edema may not be clear. Therefore, this study aims to investigate the practical benefits of delayed imaging by comparing the image from a patient with brain metastasis before a contrast enhancement and the image 10 minutes after a contrast enhancement. Materials and methods: Of the 92 people who underwent MRI brain metastases in suspected patients 13 people in three patients there is no video to target the 37 people confirmed cases, and motion artifacts brain metastases in our hospital June-December 2013, 18 people measurement position except for the three incorrect patient (male: 11 people, female: 7 people, average age: 60 years) in the target, test equipment, 3.0T MR System (ACHIEVA Release, Philips, I was 8ChannelSENSE Head Coil use Best, and the Netherlands). TR 11000 ms, TE 125 ms, TI2800 ms, Slice Thickness 5 mm, gap 5 mm, is a Slice number 21, the parameters of the 3D FFE, T2 FLAIR variable that was used to test, TR 8.1 ms, TE 3.7 ms, Slice number 240 I set to. The experiment was conducted by acquiring the FLAIR prior to contrast enhancement (heretofore referred to as Pre FLAIR), and acquiring the 3D FFE CE five minutes after the contrast enhancement, and recomposing the images in an axial plane of S/T 3mm, G 0mm (heretofore referred to as MPR TRA CE). Using the FLAIR 10 minutes after the contrast enhancement (heretofore referred to as Post FLAIR) and Pi-View, a retrospective study was conducted. Using MRIcro on the image of a patient confirmed for his diagnosis, the images before and after the contrast media, as well as the CNR and SNR of the MPR TRA CE images of the lesion and the site absent of lesion were compared and analyzed using a one-way analysis of variance. Results: CNR for Pre FLAIR and Post FLAIR were 34.35 and 60.13, respectively, with MPR TRA CE at 23.77 showing no significant difference (p<0.050). Post-experiment analysis shows a difference between Pre FLAIR and Post FLAIR in terms of CNR (p<0.050), but no difference in CNR between Post FLAIR and MPR TRA CE (p>0.050), indicating that the contrast media had an effect only on Pre FLAIR and Post FLAIR. The SNR for the normal site Pre FLAIR was 106.43, and for the lesion site 140.79. Post FLAIR for the normal site was 107.79, and for the lesion site 167.91. MPR TRA CE for the normal site was 140.23 and for the lesion site 183.19, showing significant difference (p<0.050), and post-experiment analysis shows that there was a difference in SNR only on the lesion sites for Pre FLAIR and Post FLAIR (p<0.050). There was no difference in SNR between the normal site and lesion site for Post FLAIR and MPR TRA CE, indicating no effect from the contrast media (p>0.050). Conclusions: This experiment shows that Post FLAIR has a higher contrast than Pre FLAIR, and a higher SNR for lesions, It was not not statistically significant and MPR TRA CE but CNR came out high. Inspection of post-contrast which is used in a high magnetic field is frequently used images of 3D T1 but, since the signal of the contrast medium and the blood flow is included, this method can be diagnostic accuracy is reduced, it is believed that when used in combination with Post FLAIR, and that can provide video information added to the diagnosis of brain metastases.

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뇌종야의 관류 자기공명영상: 예비보고 (Perfusion MR Imaging of the Brain Tumor: Preliminary Report)

  • 김홍대;장기현;성수옥;한문희;한만청
    • Investigative Magnetic Resonance Imaging
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    • 제1권1호
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    • pp.119-124
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    • 1997
  • 목적: 뇌종양에서 자기공명 뇌혈류량지도(MR cerebral blood volume map)의 유용성을 평가하고자 하였다. 대상 및 방법: 15예의 두개강내 종괴(다형성 교모세포종 2예, 저등급 교종 3예, 뇌농양 2예, 뇌수막종 3예, 신경세포종 1예, 배아종 1예, 방사선괴사 1예, 전이암 1예) 에서 관류 MR영상을 수술전에 시행하였다. 환자의 평균연령은 42세였고(22-68), 남자 10명, 여자 5명이었다. MR영상기기는 1.5T unit(Signa, GE Medical Systems, Milwaukee, Wisconsin)를 사용하였다. 조영제 주입후 조영제의 일차 통과시 나타나는 자화율을 얻었다. (조영제는 최오의 MR 촬영시작후 10초부터 시작하여, 총량 15cc 의 Gadopentate dimeglumine(Magnevist)를 약 2ml/sec의 속도로 손으로 주입하였다). 각 환자마다 160초 동안 6 slice에서 slice당 80 image씩 총 480 image를 얻었으며 interleaved single shot gradient EPI기법을 사용하였다. 영상변수는 TR 2000ms, TE 50ms, FOV $240{\times}240mm,{\;}matrix{\;}size{\;}128{\times}128$, slice thickness/gap 5/ 2.5mm, flip angle $90^{\circ}$로 하였다. 얻은 영상데이터는 GE workstation으로 전송한 후, 자체적으로 개발한 software에 의해 각 kvoxel마다 시간경과에 따른 신호크기의 로그변화곡선(${\Delta}R2^{*}=-In(S/S_0$)의 적분값을 구하여 국소뇌혈류량(rCBV)영상을 구성하였다. 이의 시각적 해석을 용이하게 하기 위해 정상 뇌백질의 관류정도를 기준으로 하여 상대적인 RGB 색수치로 변환하여, color rCBV map을 얻었다. 관류의 정도와 조영증강정도를 중심으로 관류 MR 영상소견과 조직학적 소견을 관련지어 분석하였다. 결과: 조영증강 T1강조MR영상에서 환상조영증강을 보이는 다형성 교보세포종 2예에서는 변연부 외륜이 고관류를, 중심부의 괴사부위는 저관류로 나타났다. 저등급 교종은 경계가 불분명한 저관류부위로 보였다. 뇌농양 2예는 변연부 외륜이 경도의 고관류를, 중심부는 저관류로 나타났다. 뇌수막종은 미만성의 균일한 중등도 혹은 고도의 고관류로 보였으며, 임파종과 배아종은 경계가 명확한 저관류부위로 나타났다. 신경세포종은 종괴\ulcorner 일부에 중등도 혹은 고도의 고관류부위가 관찰되었고, 전이암은 다수병변중 일부에서 중등도의 고관류를 보였다. 방사선괴사는 저관류부위내에 국소적 고관류부위를 보였다. 결론: 관류 MR영상은 뇌종양의 관류상태를 비교적 잘 반영하며, 조직학적 특성을 예측하는데에 도움을 주 수 있을 것으로 기대된다. 뇌종야에서의 관류MR영상의 분명한 역할을 규명하기 위해서는 앞으로 더 많은 임상적 연구가 필요할 것으로 생각된다.

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VRML을 이용한 3차원 Brain-endoscopy와 2차원 단면 영상 (3D Brain-Endoscopy Using VRML and 2D CT images)

  • 김동욱;안진영;이동혁;김남국;김종효;민병구
    • 대한의용생체공학회:학술대회논문집
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    • 대한의용생체공학회 1998년도 추계학술대회
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    • pp.285-286
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    • 1998
  • Virtual Brain-endoscopy is an effective method to detect lesion in brain. Brain is the most part of the human and is not easy part to operate so that reconstructing in 3D may be very helpful to doctors. In this paper, it is suggested that to increase the reliability, method of matching 3D object with the 2D CT slice. 3D Brain-endoscopy is reconstructed with 35 slices of 2D CT images. There is a plate in 3D brain-endoscopy so as to drag upward or downward to match the relevant 2D CT image. Relevant CT image guides the user to recognize the exact part he or she is investigating. VRML Script is used to make the change in images and PlaneSensor node is used to transmit the y coordinate value with the CT image. The result is test on the PC which has the following spec. 400MHz Clock-speed, 512MB ram, and FireGL 3000 3D accelerator is set up. The VRML file size is 3.83MB. There was no delay in controlling the 3D world and no collision in changing the CT images. This brain-endoscopy can be also put to practical use on medical education through internet.

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Low Non-NMDA Receptor Current Density as Possible Protection Mechanism from Neurotoxicity of Circulating Glutamate on Subfornical Organ Neurons in Rats

  • Chong, Wonee;Kim, Seong Nam;Han, Seong Kyu;Lee, So Yeong;Ryu, Pan Dong
    • The Korean Journal of Physiology and Pharmacology
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    • 제19권2호
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    • pp.177-181
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    • 2015
  • The subfornical organ (SFO) is one of circumventricular organs characterized by the lack of a normal blood brain barrier. The SFO neurons are exposed to circulating glutamate ($60{\sim}100{\mu}M$), which may cause excitotoxicity in the central nervous system. However, it remains unclear how SFO neurons are protected from excitotoxicity caused by circulating glutamate. In this study, we compared the glutamate-induced whole cell currents in SFO neurons to those in hippocampal CA1 neurons using the patch clamp technique in brain slice. Glutamate ($100{\mu}M$) induced an inward current in both SFO and hippocampal CA1 neurons. The density of glutamate-induced current in SFO neurons was significantly smaller than that in hippocampal CA1 neurons (0.55 vs. 2.07 pA/pF, p<0.05). To further identify the subtype of the glutamate receptors involved, the whole cell currents induced by selective agonists were then compared. The current densities induced by AMPA (0.45 pA/pF) and kainate (0.83 pA/pF), non-NMDA glutamate receptor agonists in SFO neurons were also smaller than those in hippocampal CA1 neurons (2.44 pA/pF for AMPA, p<0.05; 2.34 pA/pF for kainate, p< 0.05). However, the current density by NMDA in SFO neurons was not significantly different from that of hippocampal CA1 neurons (1.58 vs. 1.47 pA/pF, p>0.05). These results demonstrate that glutamate-mediated action through non-NMDA glutamate receptors in SFO neurons is smaller than that of hippocampal CA1 neurons, suggesting a possible protection mechanism from excitotoxicity by circulating glutamate in SFO neurons.

5-Hydroxytryptamine Inhibits Glutamatergic Synaptic Transmission in Rat Corticostriatal Brain Slice

  • Cho, Hyeong-Seok;Choi, Se-Joon;Kim, Ki-Jung;Lee, Hyun-Ho;Kim, Seong-Yun;Cho, Young-Jin;Sung, Ki-Wug
    • The Korean Journal of Physiology and Pharmacology
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    • 제9권5호
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    • pp.255-262
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    • 2005
  • Striatum is involved in the control of movement and habitual memory. It receives glutamatergic input from wide area of the cerebral cortex as well as an extensive serotonergic (5-hydroxytryptamine, 5-HT) input from the raphe nuclei. In our study, the effects of 5-HT on synaptic transmission were studied in the rat corticostriatal brain slice using in vitro whole-cell recording technique. 5-HT inhibited the amplitude as well as frequency of spontaneous excitatory postsynaptic currents (sEPSC) significantly, and neither ${\gamma}-aminobutyric$ acid (GABA)A receptor antagonist bicuculline (BIC), nor $N-methyl-_{D}-aspartate$ (NMDA) receptor antagonist, $_{DL}-2-amino-5-phosphonovaleric$ acid (AP-V) could block the effect of 5-HT. In the presence non-NMDA receptor antagonist, 2,3-dioxo-6-nitro-1,2,3,4-tetrahydrobenxo[f] quinoxaline-7-sulfonamide (NBQX), the inhibitory effect of 5-HT was blocked. We also figured out that 5-HT change the channel kinetics of the sEPSC. There was a significant increase in the rise time during the 5-HT application. Our results suggest that 5-HT has an effect on both pre- and postsynaptic site with decreasing neurotransmitter release probability of glutamate and decreasing the sensitivity to glutamate by increasing the rise time of non-NMDA receptor mediated synaptic transmission in the corticostriatal synapses.