title: Sucrase-isomaltase gene expression along crypt-villus axis of human small intestine is regulated at level of mRNA abundance.
abstract: The mucosal lining of the small intestine is a complex epithelium that is continually renewed by division of a stem cell population located in intestinal crypts, migration of daughter cells along the villus, and, finally, extrusion of senescent cells into the lumen. The majority of cells in both crypt and villus cell compartments are enterocytes that acquire differentiated functions as they migrate out of the crypt. Sucrase-isomaltase (SI) is an enterocyte-specific, brush-border enzyme that has little activity in crypt cells and maximal activity in low and mid villus cells. The mechanism by which enterocytes acquire SI enzymatic activity as they move from crypt to villus is controversial. In this study we examined the distribution of SI mRNA along the crypt-villus axis of human small intestine using isolated epithelial cells and in situ hybridization. A complementary DNA to the 5' portion of the human SI mRNA was amplified and cloned using the polymerase chain reaction. Hybridization analysis of RNA extracted from human intestinal epithelial cells showed that the cloned cDNA recognized a single 6.5-kb mRNA. In situ hybridization of duodenal biopsy specimens was performed using a single-stranded RNA probe derived from this cDNA. This analysis showed that there was little SI mRNA in crypt cells and appearance of mRNA in enterocytes located at the crypt-villus junction. The mRNA levels were maximal in lower and mid villus cells with decreased levels noted in villus tip cells. These results are identical to those previously described in rat intestine and suggest that expression of the SI gene as enterocytes emerge from intestinal crypts is regulated primarily at the level of mRNA accumulation. Study of SI gene regulation may provide a useful model to investigate the mechanisms that regulate enterocyte-specific gene expression and intestinal differentiation.
authors.affiliation: Department of Internal Medicine, University of Michigan Medical School, Ann Arbor.
authors.forename: P G
authors.initials: PG
authors.lastname: Traber
authors.forename: L
authors.initials: L
authors.lastname: Yu
authors.forename: G D
authors.initials: GD
authors.lastname: Wu
authors.forename: T A
authors.initials: TA
authors.lastname: Judge
mesh_terms: D000595:Amino Acid Sequence; D015139:Blotting, Southern; D002458:Cell Fractionation; D015870:Gene Expression / Q000502:physiology*; D006801:Humans; D007421:Intestine, Small / Q000378:metabolism / Q000502:physiology*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000378:metabolism*; D013394:Sucrase-Isomaltase Complex / Q000235:genetics*
chemical_list: D012333:RNA, Messenger; D013394:Sucrase-Isomaltase Complex