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Three-Dimensional Imaging and Gene Expression Analysis Upon Enzymatic Isolation of the Tongue Epithelium


Meisel, Christian T; Pagella, Pierfrancesco; Porcheri, Cristina; Mitsiadis, Thimios A (2020). Three-Dimensional Imaging and Gene Expression Analysis Upon Enzymatic Isolation of the Tongue Epithelium. Frontiers in Physiology:11:825.

Abstract

The tongue is a complex organ involved in a variety of functions such as mastication, speech, and taste sensory function. Enzymatic digestion techniques have been developed to allow the dissociation of the epithelium from the connective tissue of the tongue. However, it is not clear if the integrity and three-dimensional architecture of the isolated epithelium is preserved, and, furthermore if this tissue separation technique excludes its contamination from the mesenchymal tissue. Here, we first describe in detail the methodology of tongue epithelium isolation, and thereafter we analyzed the multicellular compartmentalization of the epithelium by three-dimensional fluorescent imaging and quantitative real-time PCR. Molecular characterization at both protein and transcript levels confirmed the exclusive expression of epithelial markers in the isolated epithelial compartment of the tongue. Confocal imaging analysis revealed that the integrity of the epithelium was not affected, even in the basal layer, where areas of active cell proliferations were detected. Therefore, the preservation of both the architecture and the molecular signature of the tongue epithelium upon enzymatic tissue separation enable further cellular, molecular and imaging studies on the physiology, pathology, and regeneration of the tongue.

Abstract

The tongue is a complex organ involved in a variety of functions such as mastication, speech, and taste sensory function. Enzymatic digestion techniques have been developed to allow the dissociation of the epithelium from the connective tissue of the tongue. However, it is not clear if the integrity and three-dimensional architecture of the isolated epithelium is preserved, and, furthermore if this tissue separation technique excludes its contamination from the mesenchymal tissue. Here, we first describe in detail the methodology of tongue epithelium isolation, and thereafter we analyzed the multicellular compartmentalization of the epithelium by three-dimensional fluorescent imaging and quantitative real-time PCR. Molecular characterization at both protein and transcript levels confirmed the exclusive expression of epithelial markers in the isolated epithelial compartment of the tongue. Confocal imaging analysis revealed that the integrity of the epithelium was not affected, even in the basal layer, where areas of active cell proliferations were detected. Therefore, the preservation of both the architecture and the molecular signature of the tongue epithelium upon enzymatic tissue separation enable further cellular, molecular and imaging studies on the physiology, pathology, and regeneration of the tongue.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > Center for Dental Medicine > Institute of Oral Biology
Dewey Decimal Classification:610 Medicine & health
Scopus Subject Areas:Life Sciences > Physiology
Health Sciences > Physiology (medical)
Uncontrolled Keywords:Physiology (medical), Physiology
Language:English
Date:21 July 2020
Deposited On:07 Oct 2020 09:39
Last Modified:23 Apr 2024 01:43
Publisher:Frontiers Research Foundation
ISSN:1664-042X
OA Status:Gold
Free access at:PubMed ID. An embargo period may apply.
Publisher DOI:https://doi.org/10.3389/fphys.2020.00825
PubMed ID:32848819
  • Content: Published Version
  • Licence: Creative Commons: Attribution 4.0 International (CC BY 4.0)