Complex N-glycans in mgat1 null preimplantation embryos arise from maternal mgat1 RNA

Ella Ioffe, Yun Liu, Pamela Stanley

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Mice with a null mutation in the Mgat1 gene lack N-acetylglucosaminyltransferase I (GlcNAc-TI; EC 2.4.1.101), and die at mid-gestation. This result suggested that development of Mgat1(-/-) blastocysts and their implantation could occur in the absence of complex and hybrid N-glycans. However, inner cell mass of all blastocysts from several Mgat1(+/-) heterozygous crosses bind the lectin E-PHA, indicating that Mgat1 null mutant blastocysts are able to synthesize complex N-glycans (Campbell et al., (1995) Glycobiology, 5, 535-543). In order to directly test this hypothesis, Mgat1(-/-) blastocysts were positively identified by polymerase chain reaction (PCR) of genomic DNA. Reverse transcriptase PCR (RT-PCR) of RNA isolated from the same blastocysts, and restriction analysis of the PCR products, revealed that Mgat1 null blastocysts contained Mgat1 RNA derived from the wild-type Mgat1 gene. Consistent with this, all 3.5 day blastocysts from five heterozygous crosses bound the lectin L-PHA, a lectin previously shown not to bind to E8.5 or E9.5 Mgat1(-/-) embryos that lack complex N-glycans (Ioffe and Stanley (1994) Proc. Natl. Acad. Sci., USA, 91, 728-732). Blastocysts of 4.5 days postcoitum (dpc) obtained by culturing 3.5 dpc blastocysts also bound L-PHA. However, mutant embryos that did not bind L-PHA were present among progeny from E5.5 onward. Therefore, the effects of the Mgat1 null mutation are not operative until sometime between implantation and E5.5, due to the continued presence of maternally derived Mgat1 mRNA in preimplantation embryos.

Original languageEnglish (US)
Pages (from-to)913-919
Number of pages7
JournalGlycobiology
Volume7
Issue number7
DOIs
StatePublished - Oct 1997

Fingerprint

Polymerase chain reaction
Blastocyst
Lectins
Polysaccharides
Mothers
RNA
Genes
RNA-Directed DNA Polymerase
Reaction products
Messenger RNA
Embryonic Structures
Blastocyst Inner Cell Mass
DNA
Glycomics
Polymerase Chain Reaction
Mutation
plants leukoagglutinins
Reverse Transcriptase Polymerase Chain Reaction
alpha-1,3-mannosyl-glycoprotein beta-1,2-N-acetylglucosaminyltransferase I
Pregnancy

Keywords

  • Blastocyst
  • Embryonic development
  • N-linked glycans
  • PCR

ASJC Scopus subject areas

  • Biochemistry

Cite this

Complex N-glycans in mgat1 null preimplantation embryos arise from maternal mgat1 RNA. / Ioffe, Ella; Liu, Yun; Stanley, Pamela.

In: Glycobiology, Vol. 7, No. 7, 10.1997, p. 913-919.

Research output: Contribution to journalArticle

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