Genetic engineering of carbohydrate biosynthetic pathways in transgenic mice demonstrates cell cycle-associated regulation of glycoconjugate production in small intestinal epithelial cells

Lynn Bry, Per G. Falk, Jeffrey I. Gordon

Research output: Contribution to journalArticlepeer-review

29 Scopus citations

Abstract

Proliferation, migration-associated differentiation, and cell death occur continuously and in a spatially well-organized fashion along the crypt- villus axis of the mouse small intestine, making it an attractive system for studying how these processes are regulated and interrelated. A pathway for producing glycoconjugates was engineered in adult FVB/N transgenic mice by expressing a human α1,3/4-fucosyltransferase (α1,3/4-FT; EC 2.4.1.65) along the length of this crypt-villus axis. The α1,3/4-FT can use lacto-N- tetraose or lacto-neo-S-tetraose core chains to generate Lewis (Le) blood group antigens Lea or Le(x), respectively, and H type 1 or H type 2 core chains to produce Leb and Le(y). Single-and multilabel immunohistochemical studies revealed that expression of the α1,3/4-FT results in production of Le(a) and Leb antigens in both undifferentiated proliferating crypt cells and in differentiated postmitotic villus-associated epithelial cells. In contrast, Le(x) antigens were restricted to crypt cells. Villus enterocytes can be induced to reenter the cell cycle by expression of simian virus 40 tumor antigen under the control of a promoter that only functions in differentiated members of this lineage. Bitransgenic animals, generated from a cross of FVB/N α1,3/4-FT with FVB/N simian virus 40 tumor antigen mice, expand the range of Le(x) expression to include villus-associated enterocytes that have reentered the cell cycle. Thus, the fucosylations unveil a proliferation-dependent switch in oligosaccharide production, as defined by a monoclonal antibody specific for the Le(x) epitope. These findings show that genetic engineering of oligosaccharide biosynthetic pathways can be used to define markers for entry into, or progression through, the cell cycle and to identify changes in endogenous carbohydrate metabolism that occur when proliferative status is altered in a manner that is not deleterious to the system under study.

Original languageEnglish
Pages (from-to)1161-1166
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume93
Issue number3
DOIs
StatePublished - Feb 6 1996

Keywords

  • differentiation
  • fucosyltransferases
  • glycoconjugate metabolism

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