【作者】 韩笑；

【导师】 吕广明；

【作者基本信息】 南通大学 ， 人体解剖与组织胚胎学， 2008， 硕士

【摘要】 第一部分犬脊髓的解剖学观察和测量目的:观察犬脊髓基本形态,测量获取犬脊髓应用解剖学数据。方法:1.通过对12只成年家犬脊髓冰冻切片进行Nissl染色,观察脊髓整体观及各脊髓节段横断面的形态; 2.测量并统计脊髓各节段横截面横径,矢状径,灰质面积和白质面积及其比值,灰质百分比和压缩比。结果:犬脊髓位于椎管内,其长度与椎管长度不一致,分为颈(C)、胸(T)、腰(L)、骶(S)、尾(Co)几个节段(C8,T13,L7,S3,Co5),二者的位置关系大体如下:C1-7与同序数椎骨的椎体相对应;C8平对C7-8椎骨之间;T1-6比同序数椎骨高1个椎体;T7-8比同序数椎骨高半个椎体;T9-L3与同序数椎骨的椎体相对应;L4-7对应第5-6腰椎体高度;骶段和尾段相当于第6-7腰椎体高度。犬脊髓全长有两处梭型膨大:颈膨大(C5-T1)和腰膨大(L4-7)。各脊髓节段横断面横径、矢状径和灰质面积、白质面积的变化趋势基本相同。各脊髓节段横径变化较矢状径大,其中C1为脊髓全长横径最大的节段,C6为脊髓全长矢状径最大的节段。在颈膨大和腰膨大部位灰、白质比、灰质百分比和压缩比均较大。结论:本实验通过测量统计脊髓各节段横截面横径,矢状径,灰、白质面积及其比值,灰质百分比和压缩比等脊髓特征性参数,为犬脊髓的损伤再生研究提供了形态学基础。第二部分犬脊髓皮质脊髓束的BDA顺行示踪及三维可视化研究目的:提供家犬皮质脊髓束在脊髓中的走行和定位,探讨数字三维重建技术在犬皮质脊髓束中的应用。方法:选取8只成年家犬,运用生物素化葡聚糖胺(biotinylated dextran amine, BDA)免疫荧光技术顺行追踪犬脊髓皮质脊髓束,对其在脊髓横切面上进行解剖定位。基于犬脊髓连续冰冻切片获得脊髓和皮质脊髓束的二维图像,在Photoshop软件环境下完成切片图像的配准、分割和灰度化,利用3D-DOCTOR 4.0软件对脊髓及皮质脊髓束进行三维重建并可视化。结果:1.大脑皮层运动区锥体细胞及其轴突标记为绿色荧光,锥体细胞轴突呈绿色条带状下行组成锥体束。在延髓锥体中可见绿色荧光标记区域,与周围结构分界明显。延髓下端锥体交叉处可见绿色荧光标记信号经延髓中央管腹侧,越边交叉至对侧,进入脊髓外侧索。脊髓颈、胸、腰、骶段可见有绿色荧光标记区域位于外侧索后部内侧,紧邻灰质后角,境界清晰。同时发现绿色荧光标记区域在脊髓外侧索下行的过程中,位置逐渐背移,至胸4节段已可见部分绿色荧光标记区域分布于脊髓后角区域,绿色荧光标记区域范围逐渐变小,在骶段仅可见少量绿色荧光标记纤维。2. 3D-DOCTOR软件重建出整个脊髓和皮质脊髓束,可以从任意角度自由观察其结构并进行表面积、体积等应用解剖学数据的测量。结论:1.运用BDA-Streptavidin-FITC顺行示踪法可以对正常家犬皮质脊髓束进行追踪定位。2.家犬皮质脊髓束自运动皮质发出,经锥体交叉至对侧脊髓外侧索,伴随灰质后角走行,下降达骶段。3.运用3D-DOCTOR软件能够重建出整个家犬脊髓,并可以从任意角度自由观察其内部结构。第三部分犬大脑皮层和皮质脊髓束电生理信号的记录和初步分析目的:通过采集犬大脑运动皮层和脊髓皮质脊髓束电生理信号,分析描述电信号的特征,为犬脊髓损伤与修复研究提供有价值的神经电生理资料。方法:在犬大脑运动皮层和脊髓插入单电极,使用神经信号采集处理系统(Cerebus System)记录犬大脑皮层和脊髓电生理信号。利用神经信号分析软件off-line Sorter、Neuroexplorer对已存储的信号文件进行波形特点的描述,包括波长、波幅、放电频率等。结果:1.在犬大脑皮层和脊髓均稳定记录到长时间连续的皮层和脊髓传导束自发放电信号。2.大脑皮层运动神经元和脊髓内传导束放电波形呈双向放电波形,波幅均为百μv级。3.利用“Neuroexplorer”神经信号分析软件对记录的文件作进一步分析,发现皮层神经元和脊髓内传导束的放电呈连续性发放。结论:1.运用铂-玻璃微电极可以对犬大脑运动皮质和脊髓内传导束进行神经电信号的采集。2.从获取的电信号特征及电极尖端插入脊髓内的位置判断脊髓内传导束的神经电信号来源于大脑运动皮质,提示为皮质脊髓束。更多还原

【Abstract】 Part I The Anatomical Observations and Measurements of Canine Spinal CordObjective: To observe the morphologic features and measure the applied anatomy data.Methods: Cross sections of canine spinal cord at each segment were prepared for histological evaluation using Nissl stain. The morphologic features of the spinal cord and its segments were observed. The transverse and sagittal diameters, cross-sectional area, gray and white area at each section were measured. The compression ratio, the ratio of the gray matter area to the cross-sectional area, and the ratio of the gray matter area to the white matter were calculated.Results: The canine spinal cord lies in the spinal tube, whose length is inconsistent with the spinal tube. The spinal cord are divided into approximately 36 segments, including 8 cervical (C), 13 thoracic (T), 7 lumbar (L), 3 sacral (S), and 5 coccygeal segments (Co). The anatomic relationship between spinal cord and vertebral body were as follows: C1-7 was at the same level of correlated vertebrae; C8 was at the level between C7-8 vertebrae; T1-6 level was one vertebra higher than the correlated vertebrae; T7-8 level was half vertebra higher than the correlated vertebrae; T9-L3 was at the same level of correlated vertebrae; L4-7 was at the level of L5-6 vertebrae; S1-Co5 was at the level of L6-7 vertebrae. In canine’s spinal cord, there are two enlargements: cervical enlargement (C5-T1) and lumbar enlargement (L4-7). The variation tendency in spinal cord segments such as the transverse, sagittal diameters, grey and white matter areas were similar. There were more variations in transverse diameters than that of sagittal diameters at each segment. The largest transverse diameter segment of the spinal cord was C1, while the largest sagittal diameters segment was C6. Cross-sectional morphologic features, such as the ratio of grey matter area to white matter area, the ratio of the gray matter area to the cross-sectional area, the compression ratio, changed obviously at the different spinal cord levels, especially at the level of cervical and lumbar enlargements.Conclusions: Through measuring and calculating the characteristic parameters of spinal cord, such as the transverse and sagittal diameters, cross-sectional area, gray and white areas, the compression ratio, the ratio of the gray matter area to the cross-sectional area, and the ratio of the gray matter area to the white matter of each spinal cord segment, we provide the morphologic basis for the research on regeneration after spinal cord injury of canine.Part II Study on BDA Anterograde tracing and the Three Dimensional Reconstruction of Corticospinal Tract in Spinal Cord of CanineObjective: To provide the localization of corticospinal tract (CST) in spinal cord of canine with three-dimensional (3D) reconstruction technique. Methods: CST on the serial cross sections of the canine’s spinal cord was located with BDA immune fluorescent anterograde tracing technique. The two-dimensional images were taken, registered, segmented and changed into gray-scale images under the Photoshop 7.0 circumstance. 3D-DOCTOR4.0 software was used to make 3D reconstruction of spinal cord, gray matter and bilateral CSTs.Results: 1. The pyramidal neuron and its axons were labeled with green fluorescence, and the green stripe-shaped pyramidal tract was observed. At the level of pyramid of medulla oblongata, green fluorescence labeled regions were distinct from the peripheral structures obviously. The signals labeled with green fluorescence crossed to the opposite side from central canal ventralis in decussation of pyramid, entered into the lateral funiculus, and then passed in the posterior part of lateral funiculus, accompanying with the posterior horn in cervical, thoracic, lumbar and sacral segments. At the level of T4 segment, the green labeled tracts were found in the posterior horn zone, and became smaller gradually. At the level of sacral segments, there were only little positive labeled tracts left. 2. The spinal cord and the CSTs can be reconstructed , observed and measured with 3D-DOCTOR software.Conclusions: 1. Through BDA-Streptavidin- FITC anterograde tracing technique, the canine CST can be located. 2. The canine CST originates from motor cortex, passing into the opposite lateral funiculus in spinal cord through pyramid decussation, and then accompanied with the posterior horn of gray matter downwards to sacral segment. 3. The spinal cord and the CSTs of canine can be reconstructed, observed and measured using 3D-DOCTOR software. Part III Preliminary Analysis on Electrophysiological Signals of Canine Cerebral Cortex and Corticospinal TractsObjective: To provide neuroelectrophysiological information for the research of spinal cord injury rehabilitation of canine. To record and analyze the physiological signals of canine cerebral cortex and CSTs.Methods: The single microelectrode was plugged into the cerebral cortex and spinal cord of canine. The neuro-electrical signals were recorded from motor cortex and CSTs with Cerebus System. The characteristics of the recorded signals including wave length, amplitude, and discharge frequency etc, were analyzed with Off-line Sorter and Neuroexplorer softwares.Results: 1. Continuously and steadily spontaneous electrophysiological signals were recorded from the motor cortex and spinal cord of canine. 2. The waveform of the signals was independent bidirectional and the wave amplitudes were at a grade of hundred microvoltage. 3. The spontaneous electrophysiological signals were paroxysmal in motor cortex and spinal cord of canine as revealed by Neuroexplorer.Conclusions: 1. Platinum-glass microelectrode can be used to record the neuroelectrophysiological signals from the motor cortex and spinal cord of canine. 2. The characteristics of the recorded signals and the regions single microelectrode plugged into in spinal cord of canine indicate that CST detected with the electro-signals in spinal cord originates from motor cortex.更多还原