Three-Dimensional Imaging and Gene Expression Analysis Upon Enzymatic Isolation of the Tongue Epithelium
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...
| প্রধান লেখক: | , , , |
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| বিন্যাস: | প্রবন্ধ |
| ভাষা: | English |
| প্রকাশিত: |
Frontiers Media S.A.
2020-07-01
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| মালা: | Frontiers in Physiology |
| বিষয়গুলি: | |
| অনলাইন ব্যবহার করুন: | https://www.frontiersin.org/article/10.3389/fphys.2020.00825/full |
| _version_ | 1828552888826200064 |
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| author | Christian T. Meisel Pierfrancesco Pagella Cristina Porcheri Thimios A. Mitsiadis |
| author_facet | Christian T. Meisel Pierfrancesco Pagella Cristina Porcheri Thimios A. Mitsiadis |
| author_sort | Christian T. Meisel |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-12T05:09:21Z |
| format | Article |
| id | doaj.art-eea05e24420f415eb9586b380d1df4c5 |
| institution | Directory Open Access Journal |
| issn | 1664-042X |
| language | English |
| last_indexed | 2024-12-12T05:09:21Z |
| publishDate | 2020-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Physiology |
| spelling | doaj.art-eea05e24420f415eb9586b380d1df4c52022-12-22T00:37:01ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2020-07-011110.3389/fphys.2020.00825535804Three-Dimensional Imaging and Gene Expression Analysis Upon Enzymatic Isolation of the Tongue EpitheliumChristian T. MeiselPierfrancesco PagellaCristina PorcheriThimios A. MitsiadisThe 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.https://www.frontiersin.org/article/10.3389/fphys.2020.00825/fulltongue epitheliumepithelial stem cellsthree-dimensional imagingkeratin14cell proliferationSox2 |
| spellingShingle | Christian T. Meisel Pierfrancesco Pagella Cristina Porcheri Thimios A. Mitsiadis Three-Dimensional Imaging and Gene Expression Analysis Upon Enzymatic Isolation of the Tongue Epithelium Frontiers in Physiology tongue epithelium epithelial stem cells three-dimensional imaging keratin14 cell proliferation Sox2 |
| title | Three-Dimensional Imaging and Gene Expression Analysis Upon Enzymatic Isolation of the Tongue Epithelium |
| title_full | Three-Dimensional Imaging and Gene Expression Analysis Upon Enzymatic Isolation of the Tongue Epithelium |
| title_fullStr | Three-Dimensional Imaging and Gene Expression Analysis Upon Enzymatic Isolation of the Tongue Epithelium |
| title_full_unstemmed | Three-Dimensional Imaging and Gene Expression Analysis Upon Enzymatic Isolation of the Tongue Epithelium |
| title_short | Three-Dimensional Imaging and Gene Expression Analysis Upon Enzymatic Isolation of the Tongue Epithelium |
| title_sort | three dimensional imaging and gene expression analysis upon enzymatic isolation of the tongue epithelium |
| topic | tongue epithelium epithelial stem cells three-dimensional imaging keratin14 cell proliferation Sox2 |
| url | https://www.frontiersin.org/article/10.3389/fphys.2020.00825/full |
| work_keys_str_mv | AT christiantmeisel threedimensionalimagingandgeneexpressionanalysisuponenzymaticisolationofthetongueepithelium AT pierfrancescopagella threedimensionalimagingandgeneexpressionanalysisuponenzymaticisolationofthetongueepithelium AT cristinaporcheri threedimensionalimagingandgeneexpressionanalysisuponenzymaticisolationofthetongueepithelium AT thimiosamitsiadis threedimensionalimagingandgeneexpressionanalysisuponenzymaticisolationofthetongueepithelium |