Gene expression profiling of intestinal epithelial cell maturation along the crypt-villus axis.
OBJECTIVE
To define the genetic reprogramming that drives intestinal epithelial cell maturation along the crypt-villus axis, enterocytes were sequentially isolated from the villus tip to the crypts of mouse small intestine.


METHODS
Changes in gene expression were assessed using 27,405-element complementary DNA microarrays (14,685 unique genes) and specific changes validated by Western blotting.


RESULTS
A total of 1113 genes differentially expressed between the crypt and villus were identified. Among these, established markers of absorptive and goblet cell differentiation were up-regulated in villus cells, whereas Paneth cell markers were maximally expressed in crypt cells. The 1113 differentially expressed genes were significantly enriched for genes involved in cell cycle progression, RNA processing, and translation (all predominantly down-regulated during maturation) and genes involved in cytoskeleton assembly and lipid uptake (predominantly up-regulated during maturation). No enrichment for apoptosis-regulating genes was observed. We confirmed that Wnt signaling was maximal in the proliferative compartment and observed a decrease in MYC and an increase in MAD and MAX expression during the maturation program. Consistent with these changes, the 1113 genes were enriched for MYC targets, establishing the importance of this network in intestinal cell maturation.


CONCLUSIONS
This database serves as a resource for understanding the molecular mechanisms of intestinal cell maturation and for dissection of how perturbations in the maturation process can lead to changes in gastrointestinal physiology and pathology, particularly intestinal tumorigenesis.
Mariadason
John M
JM
Montefiore Medical Cente, Albert Einstein Cancer Center, Bronx, New Yorrk 10467, USA. jmariada@aecom.yu.edu
Nicholas
Courtney
C
L'Italien
Kaitlin E
KE
Zhuang
Min
M
Smartt
Helena J M
HJ
Heerdt
Barbara G
BG
Yang
Wancai
W
Corner
Georgia A
GA
Wilson
Andrew J
AJ
Klampfer
Lidija
L
Arango
Diego
D
Augenlicht
Leonard H
LH
D000818:Animals; D017209:Apoptosis / Q000235:genetics; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D050976:Basic-Leucine Zipper Transcription Factors; D015153:Blotting, Western; D002453:Cell Cycle; D016922:Cellular Senescence / Q000235:genetics; D003599:Cytoskeleton / Q000502:physiology; D004268:DNA-Binding Proteins / Q000235:genetics; D020895:Enterocytes / Q000378:metabolism / Q000502:physiology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D016259:Genes, myc; D006652:Histological Techniques / Q000592:standards; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D007421:Intestine, Small / Q000378:metabolism; D050356:Lipid Metabolism; D008055:Lipids / Q000096:biosynthesis; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008871:Microvilli / Q000648:ultrastructure; D020411:Oligonucleotide Array Sequence Analysis; D014176:Protein Biosynthesis; D012313:RNA / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics; D015398:Signal Transduction / Q000502:physiology; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation; D051153:Wnt Proteins
D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D050976:Basic-Leucine Zipper Transcription Factors; D004268:DNA-Binding Proteins; D036341:Intercellular Signaling Peptides and Proteins; D008055:Lipids; C499479:Mad protein, mouse; C067850:Myc associated factor X; D012097:Repressor Proteins; D014157:Transcription Factors; D051153:Wnt Proteins; C068422:Max protein, mouse; D012313:RNA