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嗅觉上皮的快速荧光活体成像。

Rapid fluorescent vital imaging of olfactory epithelium.

作者信息

Nishijima Hironobu, Zunitch Matthew J, Yoshida Masafumi, Kondo Kenji, Yamasoba Tatsuya, Schwob James E, Holbrook Eric H

机构信息

Department of Developmental, Molecular, and Chemical Biology, Tufts University School of Medicine, Boston, MA 02111, USA.

Department of Otolaryngology Head and Neck Surgery, Harvard Medical School, Massachusetts Eye and Ear, 243 Charles Street, Boston, MA 02114, USA.

出版信息

iScience. 2022 Apr 8;25(5):104222. doi: 10.1016/j.isci.2022.104222. eCollection 2022 May 20.

Abstract

Olfactory epithelium (OE) undergoes degeneration in disorders such as age-related and post-viral olfactory dysfunction. However, methods for real-time detection of OE and assessment of total extent within the nasal cavity are currently unavailable. We identified two fluorescence probes for rapidly detecting and evaluating the entire extent of mice OE with topical application. Taking advantage of the differential expression of the enzymes cytochrome p450 (CYP) and γ-glutamyltranspeptidase (GGT) in OE relative to respiratory epithelium, we utilized the conversion of coumarin (a substrate of various CYP subtypes) and gGlu-HRMG (a substrate of GGT) by these enzymes to form metabolites with fluorescent emissions in the duct cells and sustentacular cells of neuron-containing OE. In depleted and regenerated OE model, the emission of these probes remained absent in respiratory metaplasia but appeared in regenerated OE. These substrates could be used to monitor OE degeneration and follow regenerative response to therapeutic interventions.

摘要

嗅觉上皮(OE)在诸如与年龄相关的和病毒感染后的嗅觉功能障碍等疾病中会发生退化。然而,目前尚无实时检测OE以及评估鼻腔内OE整体范围的方法。我们鉴定出两种荧光探针,通过局部应用可快速检测和评估小鼠OE的整体范围。利用细胞色素P450(CYP)和γ-谷氨酰转肽酶(GGT)在OE相对于呼吸上皮中的差异表达,我们利用这些酶对香豆素(各种CYP亚型的底物)和gGlu-HRMG(GGT的底物)的转化,在含神经元的OE的导管细胞和支持细胞中形成具有荧光发射的代谢产物。在OE耗竭和再生模型中,这些探针的发射在呼吸化生中仍然不存在,但在再生的OE中出现。这些底物可用于监测OE退化并跟踪对治疗干预的再生反应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce17/9046240/15415c4b6a78/fx1.jpg

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