TITLE

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

AUTHOR(S)
George, Michael D.; Wehkamp, Jan; Kays, Robert J.; Leutenegger, Christian M.; Sabir, Sadiah; Grishina, Irina; Dandekar, Satya; Bevins, Charles L.
PUB. DATE
January 2008
SOURCE
BMC Genomics;2008, Vol. 9, Special section p1
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
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.
ACCESSION #
38122787

 

Related Articles

  • DNA Profiling of Spermatozoa by Laser Capture Microdissection and Low Volume-PCR. Cai-xia Li; Jun-ping Han; Wen-yan Ren; An-quan Ji; Xiu-lan Xu; Lan Hu // PLoS ONE;2011, Vol. 6 Issue 8, p1 

    Genetic profiling of sperm from complex biological mixtures such as sexual assault casework samples requires isolation of a pure sperm population and the ability to analyze low abundant samples. Current standard procedure for sperm isolation includes preferential lysis of epithelial contaminants...

  • Identifying single-cell molecular programs by stochastic profiling. Janes, Kevin A.; Chun-Chao Wang; Holmberg, Karin J.; Cabral, Kristin; Brugge, Joan S. // Nature Methods;Apr2010, Vol. 7 Issue 4, p311 

    Cells in tissues can be morphologically indistinguishable yet show molecular expression patterns that are remarkably heterogeneous. Here we describe an approach to comprehensively identify co-regulated, heterogeneously expressed genes among cells that otherwise appear identical. The technique,...

  • An exfoliation and enrichment strategy results in improved transcriptional profiles when compared to matched formalin fixed samples. Mojica, Wilfrido D.; Stein, Leighton; Hawthorn, Lesleyann // BMC Clinical Pathology;2007, Vol. 7, p7 

    Background: Identifying the influence formalin fixation has on RNA integrity and recovery from clinical tissue specimens is integral to determining the utility of using archival tissue blocks in future molecular studies. For clinical material, the current gold standard is unfixed tissue that has...

  • The Use of Laser Microdissection in the Identification of Suitable Reference Genes for Normalization of Quantitative Real-Time PCR in Human FFPE Epithelial Ovarian Tissue Samples. Cai, Jing; Li, Tao; Huang, Bangxing; Cheng, Henghui; Ding, Hui; Dong, Weihong; Xiao, Man; Liu, Ling; Wang, Zehua // PLoS ONE;Apr2014, Vol. 9 Issue 4, p1 

    Quantitative real-time PCR (qPCR) is a powerful and reproducible method of gene expression analysis in which expression levels are quantified by normalization against reference genes. Therefore, to investigate the potential biomarkers and therapeutic targets for epithelial ovarian cancer by...

  • Proteomics on Fixed Tissue Specimens - A Review. Reimel, Beth Ann; Sheng Pan; May, Damon H.; Shaffer, Scott A.; Goodlett, David R.; McIntosh, Martin W.; Yerian, Lisa M.; Bronner, Mary P.; Ru Chen; Brentnall, Teresa A. // Current Proteomics;Apr2009, Vol. 6 Issue 1, p63 

    The vast majority of clinical tissue samples are formalin-fixed and paraffin-preserved. This type of preservation has been considered an obstacle to protein extraction from these tissues. However, these are the very tissue samples that have associated patient histories, diagnoses and outcomes -...

  • Sex-specific fluorescent labelling of cells for laser microdissection and DNA profiling. K. Anslinger; B. Bayer; B. Mack; W. Eisenmenger // International Journal of Legal Medicine;Jan2007, Vol. 121 Issue 1, p54 

    Abstract??Sex-specific isolation of cells from mixtures would greatly facilitate forensic casework. Thus, male and female cell mixtures were marked with a fluorescent X/Y-probe CEP X SpectrumOrange/Y SpectrumGreen DNA probe kit for fluorescence in situ hybridization, and single cells were...

  • Trans/paracellular, surface/crypt, and epithelial/subepithelial resistances of mammalian colonic epithelia. Gitter, A.H.; Bendfeldt, K.; Schulzke, J.D.; Fromm, M. // Pflugers Archiv European Journal of Physiology;Feb2000, Vol. 439 Issue 4, p477 

    The epithelial barrier function of the large intestine resides in the trans- and paracellular pathways of the surface epithelium and crypts. Conventional transmural resistance and permeability measurements, however, yield only the resistance of the whole tissue and not that of its individual...

  • Decreased galectin-3 expression during the progression of cervical neoplasia. Jeong-Won Lee; Sang Song; Jung-Joo Choi; Chel Choi; Tae-Joong Kim; Jhingook Kim; Je-Ho Lee; Byoung-Gie Kim; Duk-Soo Bae // Journal of Cancer Research & Clinical Oncology;Apr2006, Vol. 132 Issue 4, p241 

    Purpose: Galectin-3 is expressed widely in epithelial and immune cells and the level of expression varies in many cancer cells relative to the normal tissues from which they arise. We investigated whether the expression of galectin-3 is associated with the progression of cervical neoplasia....

  • 8: Verification of genomic identity of microchimeric cells. Sedlmayr, Peter; Kroneis, Thomas // Chimerism;Apr-Jun2013, Vol. 4 Issue 2, p44 

    Evidence for the presence of microchimerism in a tissue can be provided by PCR analysis of polymorphic alleles such as gene sequences coding for MHC molecules, whereas unequivocal identification of single microchimeric cells in tissue sections and biopsies is more challenging. Previously...

Share

Read the Article

Courtesy of THE LIBRARY OF VIRGINIA

Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics