Abstract

Immunohistochemistry methods are widely used in modern microscopic studies. The advantage of immunoelectron microscopy is the possibility of localization of functional proteins in the ultrastructures of the cell. However, different tissues and antigens react differently to the traditionally used fixatives. Selection of optimal conditions for the preparation of immunologically labeled samples for electron microscopic studies is largely based on empirical experience and depends not only on the type of tissue, but also on the purpose of the study. Nervous tissue combines mechanically weak (myelin) and dense (collagen, cell wall) structures, which poses the controversial task of simultaneously ensuring the fixation of the structure of the tissue and the interaction of antibodies with antigen throughout the entire thickness of the sample. This paper presents the method of immunohistochemical labeling, adapted for the study of the localization of annexin V and agrin in the ultrastructure of different parts of the mouse sciatic nerve. Obtained results revealed the preferential localization of both proteins in the myelin sheaths of nerve fibers, which indicates the possible role of these proteins in the functioning of myelin. The density of anti-agrin precipitate in the skin branch is significantly higher than in the tibial, whereas annexin V is more evenly distributed.

中文翻译：

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Annexin V和Agrin在小鼠完整坐骨神经中的定位

摘要

免疫组织化学方法被广泛用于现代显微镜研究中。免疫电子显微镜的优点是可以将功能蛋白定位在细胞的超微结构中。但是，不同的组织和抗原对传统使用的固定剂的反应不同。为电子显微镜研究准备免疫标记样品的最佳条件的选择主要基于经验，不仅取决于组织的类型，而且取决于研究的目的。神经组织结合了机械上较弱的（髓磷脂）和致密的（胶原蛋白，细胞壁）结构，这引起了争议的任务，即同时确保组织结构的固定以及抗体在整个样品整个厚度上的相互作用。本文提出了一种免疫组织化学标记方法，适用于研究小鼠坐骨神经不同部位的超微结构中膜联蛋白V和凝集素的定位。获得的结果揭示了这两种蛋白在神经纤维的髓鞘中的优先定位，这表明这些蛋白在髓鞘功能中的可能作用。皮肤分支中抗凝集素沉淀的密度显着高于胫骨，而膜联蛋白V分布更均匀。这表明这些蛋白质在髓磷脂功能中的可能作用。皮肤分支中抗凝集素沉淀的密度显着高于胫骨，而膜联蛋白V分布更均匀。这表明这些蛋白质在髓磷脂功能中的可能作用。皮肤分支中抗凝集素沉淀的密度显着高于胫骨，而膜联蛋白V分布更均匀。