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青蛙嗅觉神经上皮的免疫化学标志物。

Immunochemical markers for the frog olfactory neuroepithelium.

作者信息

Key B, Akeson R A

机构信息

Division of Basic Research, Children's Hospital Research Foundation, Cincinnati, OH 45229.

出版信息

Brain Res Dev Brain Res. 1990 Dec 1;57(1):103-17. doi: 10.1016/0165-3806(90)90190-a.

Abstract

Monoclonal antibodies (mAbs) were generated that react with the major cell types in the olfactory neuroepithelium of the frog, Rana catesbeiana. This pseudostratified epithelium consists of apical supporting cells, a middle layer of olfactory receptor neurons and a heterogeneous population of basal cells consisting of basal cells proper and globose basal cells. Both olfactory receptor neurons and globose basal cells were labelled by mAb 13-OE, which recognized the neural cell adhesion molecule NCAM. The identity of these NCAM positive cells was established by analysing regenerating olfactory epithelium and by a double-antibody labelling immunofluorescence technique. The olfactory nerve was lesioned, which induced the death of olfactory receptor neurons and the subsequent proliferation of basal cells. When the regenerating olfactory epithelium was analysed prior to the reconstitution of mature olfactory neurons, mAb 13-OE reacted specifically with globose basal cells and not the basal cells proper. Simultaneous labelling of normal olfactory epithelium with mAb 13-OE and polyclonal anti-keratin antibodies, the latter of which labels supporting cells and basal cells proper, revealed no double-labelled cells. These results further confirmed that NCAM was expressed by both globose basal cells and receptor neurons but not by other cell types within the epithelium. Additional cell types in the olfactory epithelium reacted with other new mAbs: 4-OE, 5-OE, 7-OE and 9-OE. Supporting cells were stained by mAb 4-OE. Olfactory receptor neurons and the entire population of basal cells were immunoreactive with mAb 7-OE. The cilia and knobs of receptor neurons were strongly immunoreactive with mAb 5-OE whereas mAb 9-OE selectively stained olfactory knobs and not the cilia on these chemosensory cells. These studies are a first step towards experimental approaches designed to elucidate the mechanisms underlying the unique proliferative properties of the olfactory neuroepithelium in frog.

摘要

制备了与牛蛙(Rana catesbeiana)嗅觉神经上皮中的主要细胞类型发生反应的单克隆抗体(mAb)。这种假复层上皮由顶端支持细胞、嗅觉受体神经元的中间层以及由固有基底细胞和球状基底细胞组成的异质基底细胞群体构成。嗅觉受体神经元和球状基底细胞均被单克隆抗体13 - OE标记,该抗体识别神经细胞黏附分子NCAM。通过分析再生的嗅觉上皮以及采用双抗体标记免疫荧光技术确定了这些NCAM阳性细胞的身份。损伤嗅觉神经,这会导致嗅觉受体神经元死亡以及随后基底细胞的增殖。当在成熟嗅觉神经元重建之前分析再生的嗅觉上皮时,单克隆抗体13 - OE特异性地与球状基底细胞反应,而不与固有基底细胞反应。用单克隆抗体13 - OE和多克隆抗角蛋白抗体同时标记正常嗅觉上皮,后者标记支持细胞和固有基底细胞,结果未发现双标记细胞。这些结果进一步证实NCAM由球状基底细胞和受体神经元表达,而上皮内的其他细胞类型不表达。嗅觉上皮中的其他细胞类型与其他新的单克隆抗体发生反应:4 - OE、5 - OE、7 - OE和9 - OE。支持细胞被单克隆抗体4 - OE染色。嗅觉受体神经元和整个基底细胞群体对单克隆抗体7 - OE具有免疫反应性。受体神经元的纤毛和小球对单克隆抗体5 - OE具有强烈的免疫反应性,而单克隆抗体9 - OE选择性地染色嗅觉小球,而不染色这些化学感受细胞上的纤毛。这些研究是旨在阐明蛙嗅觉神经上皮独特增殖特性背后机制的实验方法的第一步。

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