组织化学染色与免疫电镜双标记方法在检测大鼠脑干前包钦格复合体细胞色素氧化酶活性中的应用

发布时间：2018-02-24 06:18

本文关键词： 细胞色素氧化酶组织化学染色免疫电镜神经激肽受体前包钦格复合体大鼠　出处：《细胞与分子免疫学杂志》2017年09期 　论文类型：期刊论文

【摘要】：目的研究脑干前包钦格复合体(pre-B9t C)细胞色素氧化酶(COX)的活性变化。方法采用COX的组织化学染色与pre-B9t C的标记物神经激肽1受体(NK1R)纳米金-银加强免疫组织化学染色双标法,进行pre-B9t C特定神经核团尤其是不同亚细胞结构的COX活性检测,并进行COX活性的半定量分析。结果光镜下NK1R免疫反应性(NK1R-ir)产物主要分布于pre-B9t C神经细胞膜,清晰勾勒神经元轮廓,COX组织化学染色呈棕色,弥漫表达于NK1R-ir神经元胞质及突起内。电镜下NK1R-ir金颗粒主要分布于pre-B9t C神经元胞体和树突膜内表面,胞质内也可见金颗粒标记。pre-B9t C神经元不同亚细胞结构(胞体、轴突终末、树突)线粒体的形状和分布均有不同,在胞体和轴突终末线粒体多为圆形和椭圆形,在树突则以长杆状线粒体多见,胞体线粒体多为管泡状嵴,而板层嵴线粒体以轴突终末和树突多见。轴突终末内线粒体多聚集分布,树突内线粒体多近胞膜散在分布,但在突触部位,线粒体分布密集,且局部膨大接近突触后膜,这与突触的信息传递功能需要大量ATP产生密切相关。COX活性反应产物分布于线粒体嵴上,线粒体嵴电子密度不同表明COX活性不同。轴突终末和树突线粒体COX活性明显高于胞体。结论 pre-B9t C神经元不同亚细胞结构能量代谢不同,其中轴突终末和树突线粒体COX活性明显高于胞体,尤其突触部位线粒体分布密集,且表达高COX活性。将COX组织化学技术和免疫电镜技术相结合,为检测区域性COX活性,尤其是区域内不同亚细胞结构的COX活性提供了新方法。
[Abstract]:Objective to study the changes of cytochrome oxidase (COXX) activity in pre-B9t C (pre-B9t C) brain-stem cells. Methods COX histochemical staining and pre-B9t C labeled neurokinin 1 receptor NK1R nano-gold and silver enhanced immunocytochemical staining were used. The activity of COX in specific nuclei of pre-B9t C, especially in different subcellular structures, and semi-quantitative analysis of COX activity were carried out. Results the products of NK1R immunoreactive NK1R-irwere mainly distributed in the membrane of pre-B9t C nerve cells under light microscope. The histochemical staining of Cox was brown and distributed in the cytoplasm and processes of NK1R-ir neurons. Under electron microscope, NK1R-ir gold granules were mainly distributed in the cell body of pre-B9t C neurons and the inner surface of dendritic membrane. It was also found that the shape and distribution of mitochondria in pre-B9t C neurons (cell body, axon terminal, dendrite) were different in cytoplasm. The mitochondria were round and oval in the cytoplasm and axon terminal mitochondria. In dendrites, the long rod mitochondria are more common, the mitochondria in the cell body are tubular vesicular cristae, while the lamellar mitochondria are axon terminals and dendrites, the mitochondria in the dendritic terminals are mostly clustered, and the mitochondria in the dendrites are dispersed near the cellular membrane. However, at the synaptic site, the mitochondria are densely distributed and the local swelling is close to the postsynaptic membrane, which is closely related to the production of a large number of ATP activity products on the mitochondrial crest, which is closely related to the information transmission function of the synapse. The mitochondrial crest electron density indicated that COX activity was different. The COX activity of axon terminal and dendritic mitochondria was significantly higher than that of cell body. Conclusion the energy metabolism of pre-B9t C neurons is different in different subcellular structures. The COX activity of axon terminal and dendritic mitochondria was significantly higher than that of the cell body, especially the mitochondria in synaptic sites were densely distributed and expressed high COX activity. COX histochemistry and immunoelectron microscopy were combined to detect the regional COX activity. In particular, the COX activity of different subcellular structures in the region provides a new method.
【作者单位】：第四军医大学神经生物学教研室;第四军医大学病理与病理生理学教研室;
【基金】：国家自然科学基金(31171102,31471097)
【分类号】：R338

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