Immunohistochemical and Ultrastructual Study of Brain Injury Produced by Repeated Impacts.

反复撞击造成的脑损伤的免疫组织化学和超微结构研究。

• 批准号: 09670450
• 负责人: AOKI Yasuhiro
• 金额: $ 2.11万
• 依托单位: Iwate Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670450/
• 关键词: brain injury; axonal injury; immunohistochemistry; animal experiment; cell culture; ultramicrostructure; 免疫組織化学; fluid percussion モデル; 三叉神経節; fluid percussionモデル

To investigate mechanical effect of repeated minor impact to the pathobiology of the brain, 2 experimental head injury models were designed. In vivo, adult rats were injured utilizing weight-drop device consisting of brass weight(450g) falling through a acrylic guide tube(1m). After 3 insults at an interval of 24 hours, the brain were perfused with 4% Paraformaldehyde in phosphate buffer solution and removed. Histopathological and immunohistochemical studies disclosed neuronal injury in the cerebral cortex, and brain edema in the corpus callosum. Despite the lack of diffuse axonal changes, this model would be suitable for studying neuronal, changes associated with minor head trauma. In vitro model of axonal injury using cultured cells was designed to introduce traumatic alterations on neuronal processes and to identify mechanisms responsible for the formation of focal swellings by observation with phase-contrast and transmission electron microscopes. The culture dishes were oscillated … More one-dimensionally and horizontally 1, 2 or 3 times; repeating at intervals of an hour, 5 seconds a time; on a heat controlled shaker. The amplitude and frequency of the oscillation was 40 mm and 1 Hz, respectively. The injured processes showed two forms: the terminal increase in diameter of the processes and beading depending on injured portions. The microscopic finding suggests that the stress would destroy a cytoskeletal network, and then cause the spherical deformation of the processes. Ultramicroscopically, the beads contained compactly and sparsely misaligned cytoskeletons and prominently gathered densecore vesicles. The cytoskeletal reconstruction within the regenerated growth cones was also noted in the terminal swellings and beads. The destruction of these linkers would be responsible to initiate the cytoskeletal destruction. The initiating point of the cytoskeletal alterations, however, must be confirmed using three dimensional imaging of samples without soluble proteins and immunohistochemical study. Less

为了探讨重复轻微撞击对大脑病理生物学的机械效应，设计了2种实验性头部损伤模型。在体内，使用由黄铜重物（450 g）通过丙烯酸导管（1 m）下落组成的重物下落装置对成年大鼠进行损伤。在间隔24小时的3次损伤后，用4%多聚甲醛的磷酸盐缓冲溶液灌注脑并取出。组织学和免疫组织化学研究显示大脑皮质神经元损伤，胼胝体脑水肿。尽管缺乏弥漫性轴突变化，该模型将适合于研究与轻微头部创伤相关的神经元变化。采用体外培养的神经元轴突损伤模型，通过相差显微镜和透射电镜观察，对神经元突起进行创伤性改变，并确定局灶性轴突损伤形成的机制。振荡培养皿 ...更多信息 一维和水平1、2或3次;以1小时的间隔重复，每次5秒;在热控制的振荡器上。振荡的幅度和频率分别为40 mm和1 Hz。损伤的突起表现为两种形式：突起末端直径增大和串珠状突起。显微镜观察结果表明，应力会破坏细胞骨架网络，从而导致突起的球形变形。在超微结构上，微珠含有紧密的和稀疏的错位的细胞骨架和显著聚集的致密核囊泡。再生的生长锥内的细胞骨架重建也注意到在终端selling和珠。这些连接体的破坏将负责启动细胞骨架破坏。然而，细胞骨架改变的起始点必须使用无可溶性蛋白的样品的三维成像和免疫组化研究来确认。少

项目成果

Nakayama Y, Aoki Y.: "tudies on the mechanisms responsible for formation of focal swelling on neuronal processes using a novel in-vitro model of axonal injury."Journal of Neurotrauma. (in press). (2000)

Nakayama Y，Aoki Y.：“使用新型轴突损伤体外模型研究神经元过程中局灶性肿胀形成的机制。”神经创伤杂志。

Nakayama Y, Aoki Y: "Mechanism responsible for the formation of focal swelling on injured neuronal processes: Using a novel in vitro model of axonal injury."Forensic Science International (Suppl.). (in press). (2000)

Nakayama Y，Aoki Y：“负责在受损神经元过程中形成局灶性肿胀的机制：使用轴突损伤的新型体外模型。”国际法医科学（增刊）。

Nakayama Y, Niitsu H, Aoki Y: "The ultrastructural alterations of processes of the trigeminal ganglion cell in culture by repeated acceleration and decceleration."6th Indo Pacific Congress on Legal Medicine and Forensic Sciences. 991-994 (1999)

Nakayama Y、Niitsu H、Aoki Y：“反复加速和减速培养中三叉神经节细胞过程的超微结构改变。”第六届印度太平洋法律医学和法医学大会。

Nakayama Y,Niitsu H,Aoki Y.: "The ultrastractural alterations of processes of the trigeminal ganglion cell in culture by repeated acceleration and decceleration"6th Indo Pacific Congress on Legal Medicine and Forensic Sciences. 991-994 (1999)

Nakayama Y，Niitsu H，Aoki Y.：“通过重复加速和减速培养中三叉神经节细胞过程的超结构改变”第六届印度太平洋法律医学和法医学大会。

Nakayama Y,Aoki Y.: "Mechanism responsible for the formation of focal swelling on injured neuronal processes:Using a novel in vitro model of axonal injury"Forensic Science International(Suppl.). (in press). (2000)

Nakayama Y，Aoki Y.：“损伤神经元过程中局灶性肿胀形成的机制：使用轴突损伤的新型体外模型”国际法医科学（增刊）。