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Three dimensional analysis of dendrites of the CNS neurons.

CNS 神经元树突的三维分析。

基本信息

批准号：
62480099
负责人：
HAMA Kiyoshi
金额：
$ 3.52万
依托单位：
School of Human Sciences, Waseda University (1988-1989)Okazaki National Research Institutes (1987)
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1989
项目状态：
已结题

项目摘要

We have developed a method of extracting three dimensional information quantitatively from high voltage electron microscope stereo images. To improve the accuracy of the result, we introduced into Hitachi HVEM H-1250 a rotation free zoom optical system and a side entry rotation tilt stage with a feed back control mechanism for the tilt angle. The microscope is also equipped with an image intensifier in combination with a real time image processor to reduce the beam damage to the specimen.Number, length and diameter of dendritic spines of the granule cell in the rat dentate gyrus were measured with the aid of this system. 5 m thick Golgi preparations were used for the measurement.Spine densities of 3.4, 2.3, and 2.0 per 1mu m at proximal (P), middle (m), and distal (d) portion of the dendrite respectively were obtained, which were 1.6-2.0 fold of the previous light microscopical report. Since the diameter of the dendrite decreases from proximal to distal portion, the spine densities per unit surface area of the dendrite was about equal, 0.8/mu m^2, throughout the dendritic tree.Spines of various shapes , calyx shape,mushroom shape, club shape , thin thread-like etc. were found , however the large stubby spines which were reported to be 20-40 % by light microscopy were rare, less than 2%. Mean three dimensional spine length were 1.25mu m, 1.26mu m, and 1.24 mu m at p, m, and d portions respectively, which were about 1.4 times longer than those measured in two dimensions.The surface area of the spines were measured using morphological parameters , and it was found that the total surface areas of the dendrite was doubled by the addition of the spines at each dendritic portion.It became evident that the resolution of the light microscope was not sufficient for the precise morphometry of the dendritic spine.

我们发展了一种从高压电子显微镜立体图像中定量提取三维信息的方法。为了提高测量结果的准确性，我们在日立HVEM H-1250中引入了自由旋转变焦光学系统和带有倾斜角反馈控制机构的侧入旋转倾斜台。显微镜还配备了图像增强器和实时图像处理器，以减少射线对标本的损伤。利用该系统测量了大鼠齿状回颗粒细胞树突棘的数量、长度和直径。用5米厚的高尔基体标本进行测量，在树突的近端(P)、中段(M)和远端(D)，每1亩米的脊柱密度分别为3.4、2.3和2.0，是以前光学显微镜报道的1.6-2.0倍。由于树突直径从近端到远端逐渐减小，整个树突的单位表面积上的刺密度大致相等，约为0.8个/亩m^2。树枝上有各种形状的刺，如花瓣状、蘑菇状、棒状、细丝状等，而光镜下报道为20-40%的大而粗的刺很少见，不到2%。平均三维棘长分别为1.25微米、1.26微米和1.24微米，分别是二维测量长度的1.4倍。用形态参数测量了棘突的表面积，发现每个树突部分的刺使树突的总表面积增加了一倍。可见，光学显微镜的分辨率不足以精确地测量树突棘的形态。