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| Status |
Public on Feb 28, 2008 |
| Title |
Human ileum epithelium LCMD Project |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
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, many of the mechanisms governing intestinal epithelial cell migration and the coordination of interactions with adjacent cells and the extracellular matrix are not fully understood. We have evaluated in vivo gene expression patterns of ileal epithelial cells in healthy human subjects, isolated by laser capture microdissection from either the villus epithelial or crypt cell regions of the small intestinal mucosa. Expression profiles in villus epithelium and Paneth cell lineages were determined by quantitative real-time PCR, DNA microarray, and immunohistochemistry based methods. Relative expression levels of selected epithelial biomarkers were compared between the ileum, jejunum, duodenum, colon, stomach, and esophagus. 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. 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, most notably in the ileum, in comparison to other sites along the gastrointestinal tract. Our findings provide new and important insights regarding the molecular machinery employed by small intestinal epithelial cells to mediate their function and spatial organization in vivo.
Keywords: analysis of epithelial cells from crypt or upper villus regions
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| Overall design |
Surgical specimens of human intestinal mucosa were obtained from 4 individuals undergoing surgery for colon cancer, bowel obstruction, or other non-inflammatory conditions. The samples were fixed overnight in 4% (w/v) paraformaldehyde, dehydrated in a graded alcohol series and paraffin-embedded. At least 500 epithelial cells from the crypt or upper villus regions were then captured by LCMD from unstained 6µm thick sections using a PALM MicroLaser System Total RNA was extracted for microarray-based gene expression analysis.
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| Contributor(s) |
George MD, Bevins CL |
| Citation(s) |
18457593 |
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| Submission date |
Feb 25, 2008 |
| Last update date |
Mar 12, 2014 |
| Contact name |
Michael George |
| E-mail(s) |
[email protected]
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| Organization name |
University of California, Davis
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| Department |
Medical Microbiology and Immunology
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| Street address |
451 Health Sciences Drive
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| City |
Davis |
| State/province |
CA |
| ZIP/Postal code |
95616 |
| Country |
USA |
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| Platforms (1) |
| GPL1352 |
[U133_X3P] Affymetrix Human X3P Array |
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| Samples (7)
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| Relations |
| BioProject |
PRJNA107697 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE10629_RAW.tar |
26.8 Mb |
(http)(custom) |
TAR (of CEL, CHP) |
| Processed data included within Sample table |
| Processed data provided as supplementary file |
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