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上鼻甲作为功能性人类嗅觉受体神经元的来源。

The superior turbinate as a source of functional human olfactory receptor neurons.

作者信息

Lane Andrew P, Gomez George, Dankulich Tatyana, Wang Hongyan, Bolger William E, Rawson Nancy E

机构信息

Department of Otolaryngology-Head and Neck Surgery, Johns Hopkins University School of Medicine, Baltimore, Maryland, U.S.A.

出版信息

Laryngoscope. 2002 Jul;112(7 Pt 1):1183-9. doi: 10.1097/00005537-200207000-00007.

Abstract

OBJECTIVES

The function of human olfactory receptor neurons (ORNs) remains incompletely understood, in part because of the difficulty of obtaining viable olfactory tissue for study. During endoscopic sphenoidotomy, a portion of the superior turbinate is often removed to achieve wide and safe access to the sphenoid sinus. The purpose of this study was to determine whether functional olfactory mucosa could be obtained from such superior turbinate tissue.

STUDY DESIGN/METHODS: Superior turbinate tissue was resected from 4 patients undergoing transnasal endoscopic approaches to the sphenoid sinus. The gross appearance of the turbinate mucosa was normal at the time of surgery. The specimens were placed directly into cold cell culture media and transferred to the laboratory. A portion of the mucosa was fixed and embedded for histology and immunohistochemistry. The remaining tissue was enzymatically dissociated, and the resulting cell suspension was either prepared for immediate calcium imaging or placed into cell culture. Cultured ORNs underwent calcium imaging after several weeks to assess their ability to respond to odorants.

RESULTS

Histologic analysis of superior turbinate tissue revealed the presence of patchy olfactory neuroepithelium staining positive for olfactory marker protein. Acutely dissociated ORNs were capable of generating calcium responses to odorant mixtures. ORNs could be maintained in mixed culture and retained their ability to respond to odorants.

CONCLUSIONS

Superior turbinate tissue removed during endoscopic sphenoidotomy can provide a valuable source of human olfactory neuroepithelium for functional or histologic study. Superior turbinate tissue yields stem cells and immature neurons capable of differentiating into ORNs that retain many of their functional characteristics even after growth in culture.

摘要

目的

人类嗅觉受体神经元(ORN)的功能仍未完全了解,部分原因是难以获取用于研究的存活嗅觉组织。在内镜蝶窦切开术中,通常会切除部分上鼻甲以实现对蝶窦的广泛且安全的入路。本研究的目的是确定能否从上鼻甲组织中获取功能性嗅觉黏膜。

研究设计/方法:从4例接受经鼻内镜蝶窦手术的患者中切除上鼻甲组织。手术时鼻甲黏膜的外观正常。标本直接放入冷细胞培养基中并转移至实验室。将部分黏膜固定并包埋用于组织学和免疫组织化学检查。其余组织进行酶解,所得细胞悬液要么准备立即进行钙成像,要么放入细胞培养中。培养数周后的ORN进行钙成像以评估其对气味剂的反应能力。

结果

上鼻甲组织的组织学分析显示存在散在的嗅觉神经上皮,对嗅觉标记蛋白染色呈阳性。急性解离的ORN能够对气味剂混合物产生钙反应。ORN可以在混合培养中维持,并保留其对气味剂的反应能力。

结论

在内镜蝶窦切开术中切除的上鼻甲组织可为功能性或组织学研究提供宝贵的人类嗅觉神经上皮来源。上鼻甲组织产生干细胞和未成熟神经元,这些细胞能够分化为ORN,即使在培养生长后仍保留其许多功能特征。

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