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In vivo gene expression profiling of human intestinal epithelial cells: analysis by laser microdissection of formalin fixed tissues

BACKGROUND: The small intestinal epithelium mediates vital functions of nutrient absorption and host defense. The spatial organization of the epithelial cells along the crypt-villus axis segregates them into regions of specialized function. However, the differences in transcriptional programming and...

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Autores principales: George, Michael D, Wehkamp, Jan, Kays, Robert J, Leutenegger, Christian M, Sabir, Sadiah, Grishina, Irina, Dandekar, Satya, Bevins, Charles L
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2394537/
https://www.ncbi.nlm.nih.gov/pubmed/18457593
http://dx.doi.org/10.1186/1471-2164-9-209
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author George, Michael D
Wehkamp, Jan
Kays, Robert J
Leutenegger, Christian M
Sabir, Sadiah
Grishina, Irina
Dandekar, Satya
Bevins, Charles L
author_facet George, Michael D
Wehkamp, Jan
Kays, Robert J
Leutenegger, Christian M
Sabir, Sadiah
Grishina, Irina
Dandekar, Satya
Bevins, Charles L
author_sort George, Michael D
collection PubMed
description BACKGROUND: The small intestinal epithelium mediates vital functions of nutrient absorption and host defense. The spatial organization of the epithelial cells along the crypt-villus axis segregates them into regions of specialized function. However, the differences in transcriptional programming and the molecular machinery that governs the migration, adhesion, and differentiation of intestinal epithelial cell lineages in humans remain under-explored. To increase our understanding of these mechanisms, we have evaluated gene expression patterns of ileal epithelial cells isolated by laser capture microdissection from either the villus epithelial or crypt cell regions of healthy human small intestinal mucosa. Expression profiles in villus and crypt epithelium were determined by DNA microarray, quantitative real-time PCR, and immunohistochemistry based methods. The expression levels of selected epithelial biomarkers were also compared between gastrointestinal tissues. RESULTS: Previously established biomarkers as well as a novel and distinct set of genes believed to be linked to epithelial cell motility, adhesion, and differentiation were found to be enriched in each of the two corresponding cell populations (GEO accession: GSE10629). Additionally, high baseline expression levels of innate antimicrobials, alpha defensin 5 (HD5) and regenerating islet-derived 3 alpha (Reg3A), were detected exclusively within the small bowel crypt, most notably in the ileum in comparison to other sites along the gastrointestinal tract. CONCLUSION: The elucidation of differential gene expression patterns between crypt and villus epithelial cell lineages in human ileal tissue provides novel insights into the molecular machinery that mediates their functions and spatial organization. Moreover, our findings establish an important framework of knowledge for future investigations of human gastrointestinal diseases.
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spelling pubmed-23945372008-05-23 In vivo gene expression profiling of human intestinal epithelial cells: analysis by laser microdissection of formalin fixed tissues George, Michael D Wehkamp, Jan Kays, Robert J Leutenegger, Christian M Sabir, Sadiah Grishina, Irina Dandekar, Satya Bevins, Charles L BMC Genomics Research Article BACKGROUND: The small intestinal epithelium mediates vital functions of nutrient absorption and host defense. The spatial organization of the epithelial cells along the crypt-villus axis segregates them into regions of specialized function. However, the differences in transcriptional programming and the molecular machinery that governs the migration, adhesion, and differentiation of intestinal epithelial cell lineages in humans remain under-explored. To increase our understanding of these mechanisms, we have evaluated gene expression patterns of ileal epithelial cells isolated by laser capture microdissection from either the villus epithelial or crypt cell regions of healthy human small intestinal mucosa. Expression profiles in villus and crypt epithelium were determined by DNA microarray, quantitative real-time PCR, and immunohistochemistry based methods. The expression levels of selected epithelial biomarkers were also compared between gastrointestinal tissues. RESULTS: Previously established biomarkers as well as a novel and distinct set of genes believed to be linked to epithelial cell motility, adhesion, and differentiation were found to be enriched in each of the two corresponding cell populations (GEO accession: GSE10629). Additionally, high baseline expression levels of innate antimicrobials, alpha defensin 5 (HD5) and regenerating islet-derived 3 alpha (Reg3A), were detected exclusively within the small bowel crypt, most notably in the ileum in comparison to other sites along the gastrointestinal tract. CONCLUSION: The elucidation of differential gene expression patterns between crypt and villus epithelial cell lineages in human ileal tissue provides novel insights into the molecular machinery that mediates their functions and spatial organization. Moreover, our findings establish an important framework of knowledge for future investigations of human gastrointestinal diseases. BioMed Central 2008-05-05 /pmc/articles/PMC2394537/ /pubmed/18457593 http://dx.doi.org/10.1186/1471-2164-9-209 Text en Copyright © 2008 George et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
George, Michael D
Wehkamp, Jan
Kays, Robert J
Leutenegger, Christian M
Sabir, Sadiah
Grishina, Irina
Dandekar, Satya
Bevins, Charles L
In vivo gene expression profiling of human intestinal epithelial cells: analysis by laser microdissection of formalin fixed tissues
title In vivo gene expression profiling of human intestinal epithelial cells: analysis by laser microdissection of formalin fixed tissues
title_full In vivo gene expression profiling of human intestinal epithelial cells: analysis by laser microdissection of formalin fixed tissues
title_fullStr In vivo gene expression profiling of human intestinal epithelial cells: analysis by laser microdissection of formalin fixed tissues
title_full_unstemmed In vivo gene expression profiling of human intestinal epithelial cells: analysis by laser microdissection of formalin fixed tissues
title_short In vivo gene expression profiling of human intestinal epithelial cells: analysis by laser microdissection of formalin fixed tissues
title_sort in vivo gene expression profiling of human intestinal epithelial cells: analysis by laser microdissection of formalin fixed tissues
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2394537/
https://www.ncbi.nlm.nih.gov/pubmed/18457593
http://dx.doi.org/10.1186/1471-2164-9-209
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