TY - JOUR
T1 - In vivo consequences of deleting EGF repeats 8-12 including the ligand binding domain of mouse Notch1
AU - Ge, Changhui
AU - Liu, Tongyi
AU - Hou, Xinghua
AU - Stanley, Pamela
N1 - Funding Information:
The authors gratefully acknowledge the excellent technical assistance of Wen Dong. We also thank Dr. Shaolin Shi (Mt. Sinai School of Medicine, New York) for reagents, advice and comments on the manuscript, and we thank all those who supplied mice, cell lines and probes as noted in the text. This work was supported by NIH grant RO1 NCI 95022 to PS and partial support was provided by the Albert Einstein Cancer Center grant NCI PO1 13330.
PY - 2008
Y1 - 2008
N2 - Background. Notch signaling is highly conserved in the metazoa and is critical for many cell fate decisions. Notch activation occurs following ligand binding to Notch extracellular domain. In vitro binding assays have identified epidermal growth factor (EGF) repeats 11 and 12 as the ligand binding domain of Drosophila Notch. Here we show that an internal deletion in mouse Notch1 of EGF repeats 8-12, including the putative ligand binding domain (lbd), is an inactivating mutation in vivo. We also show that maternal and zygotic Notch1lbd/lbd mutant embryos develop through gastrulation to mid-gestation. Results. Notch1lbd/lbd embryos died at mid-gestation with a phenotype indistinguishable from Notch1 null mutants. In embryonic stem (ES) cells, Notch1lbd was expressed on the cell surface at levels equivalent to wild type Notch1, but Delta1 binding was reduced to the same level as in Notch1 null cells. In an ES cell co-culture assay, Notch signaling induced by Jagged1 or Delta1 was reduced to a similar level in Notch1 lbdand Notch1 null cells. However, the Notch1lbd/lbd allele was expressed similarly to wild type Notch1 in Notch1lbd/lbd ES cells and embryos at E8.75, indicating that Notch1 signaling is not essential for the Notch1 gene to be expressed. In addition, maternal and zygotic Notch1 mutant blastocysts developed through gastrulation. Conclusion. Mouse Notch1 lacking the ligand binding domain is expressed at the cell surface but does not signal in response to the canonical Notch ligands Delta1 and Jagged1. Homozygous Notch1lbd/lbd mutant embryos die at ∼E10 similar to Notch1 null embryos. While Notch1 is expressed in oocytes and blastocysts, Notch1 signaling via canonical ligands is dispensable during oogenesis, blastogenesis, implantation and gastrulation.
AB - Background. Notch signaling is highly conserved in the metazoa and is critical for many cell fate decisions. Notch activation occurs following ligand binding to Notch extracellular domain. In vitro binding assays have identified epidermal growth factor (EGF) repeats 11 and 12 as the ligand binding domain of Drosophila Notch. Here we show that an internal deletion in mouse Notch1 of EGF repeats 8-12, including the putative ligand binding domain (lbd), is an inactivating mutation in vivo. We also show that maternal and zygotic Notch1lbd/lbd mutant embryos develop through gastrulation to mid-gestation. Results. Notch1lbd/lbd embryos died at mid-gestation with a phenotype indistinguishable from Notch1 null mutants. In embryonic stem (ES) cells, Notch1lbd was expressed on the cell surface at levels equivalent to wild type Notch1, but Delta1 binding was reduced to the same level as in Notch1 null cells. In an ES cell co-culture assay, Notch signaling induced by Jagged1 or Delta1 was reduced to a similar level in Notch1 lbdand Notch1 null cells. However, the Notch1lbd/lbd allele was expressed similarly to wild type Notch1 in Notch1lbd/lbd ES cells and embryos at E8.75, indicating that Notch1 signaling is not essential for the Notch1 gene to be expressed. In addition, maternal and zygotic Notch1 mutant blastocysts developed through gastrulation. Conclusion. Mouse Notch1 lacking the ligand binding domain is expressed at the cell surface but does not signal in response to the canonical Notch ligands Delta1 and Jagged1. Homozygous Notch1lbd/lbd mutant embryos die at ∼E10 similar to Notch1 null embryos. While Notch1 is expressed in oocytes and blastocysts, Notch1 signaling via canonical ligands is dispensable during oogenesis, blastogenesis, implantation and gastrulation.
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U2 - 10.1186/1471-213X-8-48
DO - 10.1186/1471-213X-8-48
M3 - Article
C2 - 18445292
AN - SCOPUS:44049090669
SN - 1471-213X
VL - 8
JO - BMC Developmental Biology
JF - BMC Developmental Biology
M1 - 48
ER -