AJP - Gastrointestinal and Liver Physiology, Vol 272, Issue 3 534-G544, Copyright © 1997 by American Physiological Society

ARTICLES

Increased C/EBP in fetal rat small intestine precedes initiation of differentiation marker mRNA synthesis

R. K. Montgomery, E. H. Rings, J. F. Thompson, C. C. Schuijt, K. M. Aras, V. J. Wielenga, M. J. Kothe, H. A. Buller and R. J. Grand
Division of Pediatric Gastroenterology and Nutrition, Boston Floating Hospital for Children, Tufts University School of Medicine, Massachusetts 02111, USA.

Morphogenesis, initiation of differentiation marker gene expression, and their correlation with CCAT/enhancer binding protein (C/EBP) expression were analyzed in the developing fetal rat small intestine. Expressions of mRNAs for lactase-phlorizin hydrolase (LPH), intestinal alkaline phosphatase (IALP), carbamoyl-phosphate synthetase (CPS), and three isoforms of C/EBP were simultaneously determined by Northern blot analysis from 15 to 19 days of gestation. At 17 days of gestation, prior to villus formation as demonstrated by light and electron microscopy, only CPS and C/EBPalpha, -beta, and -delta expression could clearly be detected. Both LPH and IALP mRNA were definitely detectable in proximal and middle intestine on day 18, as soon as the stratified epithelium of the early intestine had been transformed into a single layer of columnar epithelium lining villi. This distribution was confirmed by in situ hybridization for LPH mRNA. During the period of transformation when the columnar epithelium and villi were forming, no LPH or IALP mRNA was detectable in the immature distal one-third of the fetal intestine. Preceding villus morphogenesis, immunostaining demonstrated nuclear localization of C/EBPalpha protein in intestinal epithelial cells, with continued expression in all enterocytes through 19 days of gestation. Enhanced expression of C/EBPalpha mRNA and protein began 24 h prior to the initiation of the differentiation markers, suggesting that it may play a role in regulation of fetal intestinal differentiation.

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