Developmental abnormalities of glycosylphosphatidylinositol-anchor- deficient embryos revealed by Cre/IoxP system

Masami Nozaki, Kazuhito Ohishi, Naoko Yamada, Taroh Kinoshita, Andras Nagy, Junji Takeda

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One mode used to link membrane proteins to a cell membrane is by means of a special glycolipid anchor termed glycosylphosphatidylinositol (GPI). Pig-a, an X-linked gene, is involved in the first step of GPI-anchor biosynthesis. Disruption of this gene causes cessation of GPI biosynthesis on the endoplasmic reticulum, thereby leading to the absence of GPI-anchored proteins on the cell surface. We have previously reported that mice with high chimerism was never obtained from Pig-a disrupted ES cells, suggesting that GPI-anchored protein(s) may have important roles for mouse development such that the absence of GPI-anchored proteins causes a lethal effect to mice. In this study, this lethal effect has been investigated by using a conditional approach to 'knockout' the Pig-a gene. For this, mice harboring a Pig-a gene flanked by two IoxP sites (Pig-a(flox)) were mated with hCMV-Cre transgenic mice, which express Cre recombinase before implantation: The allele disruptions were identified by PCR analysis of embryo yolk sac DNA. Embryos harboring a complete disruption of Pig-a gene ceased to develop beyond the ninth day of gestation. Female embryos in which one Pig-a allele was disrupted by Cre such that only half of the cells in the embryo proper did not express GPI-anchored proteins due to random X inactivation developed until 19 days post coitum (dpc), but showed abnormal phenotypes such as insufficient closure of neural tube and cleft palate. These data further highlight the importance of GPI-anchored proteins during mouse embryonic development.

Original languageEnglish
Pages (from-to)293-299
Number of pages7
JournalLaboratory Investigation
Issue number3
Publication statusPublished - 1999
Externally publishedYes

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