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Laser capture microdissection enables cellular and molecular studies of tooth root development

Epithelial–mesenchymal interactions (EMIs) are critical for tooth development. Molecular mechanisms mediating these interactions in root formation is not well understood. Laser capture microdissection (LCM) and subsequent microarray analyses enable large scale in situ molecular and cellular studies...

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Autores principales: Sun, Jian-Xun, Horst, Orapin V, Bumgarner, Roger, Lakely, Bryce, Somerman, Martha J, Zhang, Hai
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3412663/
https://www.ncbi.nlm.nih.gov/pubmed/22422086
http://dx.doi.org/10.1038/ijos.2012.15
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author Sun, Jian-Xun
Horst, Orapin V
Bumgarner, Roger
Lakely, Bryce
Somerman, Martha J
Zhang, Hai
author_facet Sun, Jian-Xun
Horst, Orapin V
Bumgarner, Roger
Lakely, Bryce
Somerman, Martha J
Zhang, Hai
author_sort Sun, Jian-Xun
collection PubMed
description Epithelial–mesenchymal interactions (EMIs) are critical for tooth development. Molecular mechanisms mediating these interactions in root formation is not well understood. Laser capture microdissection (LCM) and subsequent microarray analyses enable large scale in situ molecular and cellular studies of root formation but to date have been hindered by technical challenges of gaining intact histological sections of non-decalcified mineralized teeth or jaws with well-preserved RNA. Here,we describe a new method to overcome this obstacle that permits LCM of dental epithelia,adjacent mesenchyme,odontoblasts and cementoblasts from mouse incisors and molars during root development. Using this method,we obtained RNA samples of high quality and successfully performed microarray analyses. Robust differences in gene expression,as well as genes not previously associated with root formation,were identified. Comparison of gene expression data from microarray with real-time reverse transcriptase polymerase chain reaction (RT-PCR) supported our findings. These genes include known markers of dental epithelia,mesenchyme,cementoblasts and odontoblasts,as well as novel genes such as those in the fibulin family. In conclusion,our new approach in tissue preparation enables LCM collection of intact cells with well-preserved RNA allowing subsequent gene expression analyses using microarray and RT-PCR to define key regulators of tooth root development.
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spelling pubmed-34126632012-09-05 Laser capture microdissection enables cellular and molecular studies of tooth root development Sun, Jian-Xun Horst, Orapin V Bumgarner, Roger Lakely, Bryce Somerman, Martha J Zhang, Hai Int J Oral Sci Original Article Epithelial–mesenchymal interactions (EMIs) are critical for tooth development. Molecular mechanisms mediating these interactions in root formation is not well understood. Laser capture microdissection (LCM) and subsequent microarray analyses enable large scale in situ molecular and cellular studies of root formation but to date have been hindered by technical challenges of gaining intact histological sections of non-decalcified mineralized teeth or jaws with well-preserved RNA. Here,we describe a new method to overcome this obstacle that permits LCM of dental epithelia,adjacent mesenchyme,odontoblasts and cementoblasts from mouse incisors and molars during root development. Using this method,we obtained RNA samples of high quality and successfully performed microarray analyses. Robust differences in gene expression,as well as genes not previously associated with root formation,were identified. Comparison of gene expression data from microarray with real-time reverse transcriptase polymerase chain reaction (RT-PCR) supported our findings. These genes include known markers of dental epithelia,mesenchyme,cementoblasts and odontoblasts,as well as novel genes such as those in the fibulin family. In conclusion,our new approach in tissue preparation enables LCM collection of intact cells with well-preserved RNA allowing subsequent gene expression analyses using microarray and RT-PCR to define key regulators of tooth root development. Nature Publishing Group 2012-04 2012-03-16 /pmc/articles/PMC3412663/ /pubmed/22422086 http://dx.doi.org/10.1038/ijos.2012.15 Text en Copyright © 2012 West China School of Stomatology http://creativecommons.org/licenses/by-nc-nd/3.0/ This work is licensed under the Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Original Article
Sun, Jian-Xun
Horst, Orapin V
Bumgarner, Roger
Lakely, Bryce
Somerman, Martha J
Zhang, Hai
Laser capture microdissection enables cellular and molecular studies of tooth root development
title Laser capture microdissection enables cellular and molecular studies of tooth root development
title_full Laser capture microdissection enables cellular and molecular studies of tooth root development
title_fullStr Laser capture microdissection enables cellular and molecular studies of tooth root development
title_full_unstemmed Laser capture microdissection enables cellular and molecular studies of tooth root development
title_short Laser capture microdissection enables cellular and molecular studies of tooth root development
title_sort laser capture microdissection enables cellular and molecular studies of tooth root development
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3412663/
https://www.ncbi.nlm.nih.gov/pubmed/22422086
http://dx.doi.org/10.1038/ijos.2012.15
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