The objective is the identification of the factors and cellular interactions which induce and modulate maturation and differentiation of the intestinal epithelium. The studies outlined in this proposal will focus on intestinal cell components defined by previously produced monoclonal antibodies which, because of their cell and/or tissue distribution in adult intestine and at different stages of development are potentially involved in those processes. New monoclonal antibodies will be prepared and characterized to further define the biological properties of the intestinal crypt cells and to detect individual cell types early in the process of commitment or differentiation. The composition, structure and biosynthesis of the newly identified antigens will be investigated by a combination of studies conducted in vivo and in culture, using intestinal epithelial cell lines and fetal rat intestinal organ cultures. The antigens recognized by the different monoclonal antibodies will be purified by affinity chromatography and characterized by slab gel electrophoresis, immunoblotting, peptide mapping, amino acid sequence determination, cell-free translation of purified intestinal mRNA, and cDNA cloning. The distribution of the various antigens in the intestinal mucosa at different stages of development and differentiation, and in cultured cells, will be determined by immunofluorescence staining and by immunoelectron-microscopy. Antibodies recognizing distinct crypt cell types will be used for a comparative study of the ultrastructure, relative position in the crypts, proliferative activity and development in fetal and newborn rats of the cells they identify. The studies should provide new insights into the process of intestinal cell differentiation, and also contribute to our knowledge of the intestinal mucosa in disease states such as sprue, inflammatory bowel diseases, interaction with luminal components, and intestinal adaptation.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK032656-07
Application #
3231035
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1982-12-01
Project End
1991-11-30
Budget Start
1988-12-01
Budget End
1989-11-30
Support Year
7
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Cornell University
Department
Type
Earth Sciences/Resources
DUNS #
City
Ithaca
State
NY
Country
United States
Zip Code
14850
Paul, E C; Quaroni, A (1993) Identification of a 102 kDa protein (cytocentrin) immunologically related to keratin 19, which is a cytoplasmically derived component of the mitotic spindle pole. J Cell Sci 106 ( Pt 3):967-81
Quaroni, A; Paul, E C; Nichols, B L (1993) Intracellular degradation and reduced cell-surface expression of sucrase-isomaltase in heat-shocked Caco-2 cells. Biochem J 292 ( Pt 3):725-34
Calnek, D; Quaroni, A (1993) Differential localization by in situ hybridization of distinct keratin mRNA species during intestinal epithelial cell development and differentiation. Differentiation 53:95-104
Quaroni, A; Nichols, B L; Quaroni, E et al. (1992) Expression and different polarity of aminopeptidase N in normal human colonic mucosa and colonic tumors. Int J Cancer 51:404-11
Rings, E H; Buller, H A; de Boer, P A et al. (1992) Messenger RNA sorting in enterocytes. Co-localization with encoded proteins. FEBS Lett 300:183-7
Calnek, D; Quaroni, A (1992) Changes in keratin expression during fetal and postnatal development of intestinal epithelial cells. Biochem J 285 ( Pt 3):939-46
Cross, H S; Quaroni, A (1991) Inhibition of sucrose-isomaltase expression by EGF in the human colon adenocarcinoma cells Caco-2. Am J Physiol 261:C1173-83
Beaulieu, J F; Quaroni, A (1991) Clonal analysis of sucrase-isomaltase expression in the human colon adenocarcinoma Caco-2 cells. Biochem J 280 ( Pt 3):599-608
Quaroni, A; Calnek, D; Quaroni, E et al. (1991) Keratin expression in rat intestinal crypt and villus cells. Analysis with a panel of monoclonal antibodies. J Biol Chem 266:11923-31
Daniele, B; Quaroni, A (1991) Effects of epidermal growth factor on diamine oxidase expression and cell growth in Caco-2 cells. Am J Physiol 261:G669-76

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