Slepecky N B, Hozza M J, Cefaratti L
Institute for Sensory Research, Syracuse University, New York 13244-1240.
J Electron Microsc Tech. 1990 Jul;15(3):280-92. doi: 10.1002/jemt.1060150307.
Immunofluorescence staining and phalloidin labeling have provided localization of actin in the sensory and supporting cells of the inner ear at the light microscopic level. However, with electron microscopy, neither actin nor actin filaments have been found in the outer hair cell body. This paper describes various techniques utilized to preserve and identify cytoplasmic actin at the ultrastructural level. Post-embedding staining of Lowicryl K4M sections, pre-embedding staining of permeabilized cells of the organ of Corti, pre-embedding staining of vibratome sections, and pre-embedding staining of permeabilized dissociated cells documented the presence of actin, but each of these techniques was best suited to localize actin in specific parts of the cell. Cytoplasmic actin was labeled when isolated cells were lightly fixed and membranes were permeabilized with detergent--conditions under which the cell ultrastructure was compromised. Under conditions of optimal fixation, cytoplasmic filaments embedded in the dense granular matrix of the hair cell cytoplasm were observed.
免疫荧光染色和鬼笔环肽标记已在光学显微镜水平上确定了肌动蛋白在内耳感觉细胞和支持细胞中的定位。然而，在电子显微镜下，在外毛细胞体中未发现肌动蛋白或肌动蛋白丝。本文描述了用于在超微结构水平上保存和鉴定细胞质肌动蛋白的各种技术。Lowicryl K4M切片的包埋后染色、柯蒂氏器通透细胞的包埋前染色、振动切片机切片的包埋前染色以及通透解离细胞的包埋前染色都证明了肌动蛋白的存在，但这些技术中的每一种都最适合于在细胞的特定部位定位肌动蛋白。当分离的细胞轻度固定并用去污剂通透膜时，细胞质肌动蛋白被标记——在这种条件下细胞超微结构受到损害。在最佳固定条件下，观察到嵌入毛细胞细胞质致密颗粒基质中的细胞质细丝。
