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Glycobiology vol 5 no 5 pp. 535-543, 1995
© 1995

research-article

Complex asparagine-linked oligosaccharides in Mgat1-null embryos

Robert M. Campbell, Martina Metzler1, Maria Granovsky2, James W. Dennis2 and Jamey D. Marth3

Howard Hughes Medical Institute and Division of Cellular and Molecular Medicine, 9500 Gilman Drive 0625, University of California San Diego La Jolla, CA 92093, USA
1The Biomedical Research Centre, 2222 Health Sciences Mall Vancouver, BC, V6T 1Z3, Canada
2Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University Avenue Toronto, Ontario, M5G 1X5, Canada

3To whom correspondence should be addressed

Received on March 30, 1995; revised on May 8, 1989; accepted on May 8, 1995

To investigate the developmental role of complex N-linked oligosaccharides, we previously inactivated the mouse Mgat1 gene which encodes UDP-N-acetylglucosamine: {alpha}-3-D-mannoside ß-1,2-N-acetylglucosaminyltransferase I (GlcNAc-TI). Mgat1-null embryos developed morphogenic abnormalities by embryonic day (E) 9.5 and failed to survive beyond E10.5. Prior to E8.5, mutant and wild-type embryos were phenotypically indistinguishable, raising the unexpected possibility that earlier embryonic development may not require complex N-glycans. We have now used in situ RNA hybridization to assess the temporal and spatial pattern of Mgat1 expression in normal embryos, and lectin histochemistry to determine whether Mgat1 -null embryos lack complex N-glycans at pre-E9.5 developmental stages. In situ RNA analysis indicated that Mgat1 transcripts normally increase dramatically between E7.0 and E9.5, 1–2 days prior to the death of mutant embryos. However, apparently normal levels of complex N-glycans were observed in E3.5 pre-implantation Mgat1-null embryos prior to declining to undetectable levels by E7.5. Complex N-glycans were not observed in E7.5-E9.5 Mgat1-null embryos with the notable exception of vesicular structures within cells of the visceral extra-embryonic endoderm, perhaps reflecting the ability of these cells to take up and transport maternally derived glycoproteins. Mgat1-null embryos appear to complete pre-implantation development in the presence of maternally derived complex N-glycaas, and may die at later stages, post E7.5, when a requirement for embyronically derived complex N-glycans arises.

complex N-linked oligosaccharides embryogenesis gene targeting


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