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嗅觉和鼻呼吸上皮,以及通过快速冷冻冷冻置换和Lowicryl K11M包埋可视化的叶状味蕾。超微结构和初步细胞化学研究。

Olfactory and nasal respiratory epithelia, and foliate taste buds visualized with rapid-freeze freeze-substitution and Lowicryl K11M embedding. Ultrastructural and initial cytochemical studies.

作者信息

Menco B P

机构信息

Department of Neurobiology and Physiology, Northwestern University, Evanston, IL 60208-3520.

出版信息

Scanning Microsc. 1989 Mar;3(1):257-72.

PMID:2740865
Abstract

Rat olfactory and respiratory epithelia and Rhesus monkey taste buds were studied with rapid-freeze, acetone/0.1% uranyl acetate freeze-substitution and low-temperature Lowicryl K11M embedding, usually in the absence of other chemical fixation and cryoprotection procedures. Ultrastructural features of mucus, cytoplasm, including cytoskeletons, and membranes were better retained than with conventional methods. Some major examples: The mucus of the olfactory epithelium consisted of a single layer; that of the respiratory epithelium had an electron-opaque sol layer surrounding cilia and microvilli below a thin laminated electron-lucent gel layer. Taste-bud pores displayed a foam-like opaque secretory product, resembling the contents of secretory granules within Type I taste-bud cells. The electron-opacity of cytoplasmic matrices sometimes obscured features such as radial spokes of respiratory cilia. Membranes had smooth outlines; those of olfactory receptor cell cilia were more electron-opaque than those of olfactory supporting cell microvilli and respiratory cilia. Membranous monolayers of many respiratory cilia across large arrays often partially split apart, all in the same direction. The space between those monolayers contained an electron-lucent substance. Preliminary cytochemistry on olfactory and nasal respiratory epithelial samples with the lectin Concanavalin A (Con A) and antibodies against olfactory marker- and odorant-binding proteins and, in taste only, the sweet-tasting protein thaumatin, were sufficiently successful to warrant further endeavors.

摘要

对大鼠嗅觉和呼吸上皮以及恒河猴味蕾进行了研究,采用快速冷冻、丙酮/0.1%醋酸铀冷冻置换和低温Lowicryl K11M包埋,通常不进行其他化学固定和冷冻保护程序。与传统方法相比,黏液、细胞质(包括细胞骨架)和膜的超微结构特征得到了更好的保留。一些主要例子如下:嗅觉上皮的黏液由单层组成;呼吸上皮的黏液在薄的分层电子透明凝胶层下方有一个围绕纤毛和微绒毛的电子不透明溶胶层。味蕾孔显示出泡沫状不透明分泌产物,类似于I型味蕾细胞内分泌颗粒的内容物。细胞质基质的电子不透明性有时会掩盖诸如呼吸纤毛的径向辐条等特征。膜具有光滑的轮廓;嗅觉受体细胞纤毛的膜比嗅觉支持细胞微绒毛和呼吸纤毛的膜电子密度更高。横跨大阵列的许多呼吸纤毛的膜单层常常在同一方向上部分分开。这些单层之间的空间含有一种电子透明物质。用凝集素伴刀豆球蛋白A(Con A)以及针对嗅觉标记物和气味结合蛋白的抗体对嗅觉和鼻呼吸上皮样本进行的初步细胞化学研究,仅在味觉方面对甜味蛋白奇异果甜蛋白进行的研究取得了足够的成功,值得进一步努力。

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