首页|大鼠海马结构有髓神经纤维体视学方法的建立

大鼠海马结构有髓神经纤维体视学方法的建立

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目的:建立定量研究海马结构及其内有髓神经纤维的无偏体视学方法. 方法:选择6 mo龄正常雌性Long-Ev-ans大鼠5只,运用卡瓦列里原理计算海马结构的总体积.根据随机抽样原则在海马结构内切取5~6个组织块,用"球切法"保证满足三维空间内各向同性抽样原则后,切取60 um厚的超薄切片,在透射电子显微镜下从每张切片随机拍摄15个视野.使用无偏计数框计算海马结构内有髓神经纤维的长度密度;使用等距测点框计算海马结构内有髓神经纤维的体积密度.用长度密度,体积密度分别乘以海马结构的体积,获得海马结构内有髓神经纤维的总长度和总体积.测量与纤维断面的最长轴相垂直的所有横轴中最长的一条横径为其直径. 结果:得到大鼠海马结构的平均体积为(85.9 ±14.0)mm3;海马结构内有髓神经纤维的总长度为(9.1±3.8)km;总体积为(4.3 ±1.2)mm3;平均直径为(0.7 ±0.1)μm. 结论:本研究建立的体视学方法可为定量研究正常老年及各种神经系统退行性疾病状态下海马结构内有髓神经纤维的改变提供依据.
Establishment of a stereological method for myelinated fibers in hippocampal formation of rat brain
AIM: To establish a stereological method for the quantitative studies of hippocampal formation and myelinated fibers in the hippocampal formation. METHODS: Five 6-month female Long-Evans rats were used. The volume of hippocampal formation was estimated using the Cavalieri's principle. Five or six tissue blocks were obtained from the hippocampal formation in a uniform random fashion. Isotropic, uniform and random (IUR) sections were ensured by isector technique and one 60 nm-thicknesses section was cut from each block. From each section, 15 fields of vision were randomly photographed using transmission electron microscope. The length density of the myelinated fibers in the hippocampal formation was estimated using an unbiased counting frame, the volume density of the myelinated fibers in the hippocampal formation was estimated using equidistant point counting grid and the total length and total volume of the myelinated fibers in the hippocampal formation were estimated by multiplying the length density and volume density by the total volume of hippocampal formation, respectively. The diameter of fibers sampled with the unbiased counting frame was estimated by measuring the profile diameter perpendicular to its longest axis. RESULTS: The average volume of rat hippocampal formation was (85.9 ± 14.0)mm3 and the total length of the myelinated fibers in the hippocampal formation was (9.1 ± 3.8) km. The total volume of the myelinated fibers in the hippocampal formation was (4.3 ± 1.2) mm3 and the average diameter of the myelinated fibers in the hippocampal formation was (0.74 ± 0.07) μm. CONCLUSION: The method established in this study provides a useful tool for the quantitative studies of the myelinated fiber changes of hippocampal formation in normal aging process and in various neurodegenerative diseases.

hippocampal formationmyelinated fibersratstereoiogy

卢伟、黄春霞、杨妹、李琛、张伟、徐瑲、唐勇

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重庆医科大学,第一附属医院神经内科,重庆,400016

重庆医科大学,生理学教研室,重庆,400016

重庆医科大学,组织胚胎学教研室,重庆,400016

海马结构 有髓神经纤维 大鼠 体视学

国家自然科学基金国家自然科学基金重庆市自然科学基金重庆市自然科学基金

3044008230572075CSTC2005BB5034

2009

第四军医大学学报
第四军医大学

第四军医大学学报

CSTPCDCSCD北大核心
影响因子:0.599
ISSN:1000-2790
年,卷(期):2009.30(18)
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