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辣根过氧化物酶标记神经元的超微结构:两种灵敏技术的比较。

Ultrastructure of HRP-labelled neurons: a comparison of two sensitive techniques.

作者信息

Mawe G M, Bresnahan J C, Beattie M S

出版信息

Brain Res Bull. 1983 Apr;10(4):551-8. doi: 10.1016/0361-9230(83)90154-5.

Abstract

Rat sciatic nerves were transected and placed in a solution of HRP. Following perfusion, L4-6 spinal segments and sensory ganglia were cut into 60 micron sections. Alternate sections were processed with the glucose oxidase (GOD) or the benzidine dihydrochloride (BDHC) procedure. Sections were then either mounted on glass slides for light microscopic (LM) examination or embedded in epoxy resin for sequential light and electron microscopic (EM) analysis. At the LM level, GOD reaction product was granular and confined to the soma and proximal dendrites of motoneurons, while BDHC reacted tissue was characterized by a diffuse labelling of the cytoplasm, extending as far as tertiary dendrites. At the EM level, neurons in tissue processed with the GOD technique contained lysosomal and vesiculotubular elements that were extremely electron dense. Label in BDHC reacted tissue appeared as dense patches of reaction product dispersed throughout the cytoplasm. Possible reasons for the qualitative differences in the reaction product provided by the two techniques as well as suggested applications of the techniques are discussed.

摘要

将大鼠坐骨神经切断并置于辣根过氧化物酶(HRP)溶液中。灌注后,将L4 - 6脊髓节段和感觉神经节切成60微米厚的切片。每隔一张切片用葡萄糖氧化酶(GOD)法或盐酸联苯胺(BDHC)法处理。然后将切片要么安装在载玻片上进行光学显微镜(LM)检查,要么包埋在环氧树脂中进行连续的光学和电子显微镜(EM)分析。在光学显微镜水平,GOD反应产物呈颗粒状,局限于运动神经元的胞体和近端树突,而BDHC反应组织的特征是细胞质弥漫性标记,延伸至三级树突。在电子显微镜水平,用GOD技术处理的组织中的神经元含有溶酶体和泡管状成分,这些成分电子密度极高。BDHC反应组织中的标记物表现为分散在整个细胞质中的致密反应产物斑块。讨论了两种技术提供的反应产物质量差异的可能原因以及这些技术的建议应用。

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