To understand the mechanisms by which the mouse epidermal growth factor receptor (EGF-R) produces morphological changes during mammalian development, a null allele (Egfr-m1Cwr) at the locus encoding the receptor was created. Surprisingly, three dramatically different phenotypes were observed depending on genetic background. Inner cell masses of Egfr-m1Cwr homozygotes on a CF-1 genetic background died at implantation. When left co-isogenic on a 129/Sv genetic background, Egfr-m1Cwr homozygotes died at mid-gestation from apparent placental failure. In contrast, the same mutant allele resulted in a peri-natal lethality when bred to homozygosity on a CD- l or C57BL/6 background. Reciprocal uterine transfers between strains showed that the resulting phenotypes were specific to the genetic background of the embryo and were not influenced by the genetic background of the host uterus. These findings have major implications regarding previous work on Egfr since virtually all experimental observations have been made within the context of undefined genetic backgrounds. To understand how these genes are able to compensate for the absence of EGF- R, a genetic analysis is proposed to determine the number and map positions of those genes involved. Appropriate statistical methods are described for the unique situation of exclusion mapping for a modifier(s) allowing for survival beyond a distinct time point. The ultimate goal is cloning of the genes responsible for the effect. A second step toward understanding the biological nature of the variable phenotypes is to determine whether the peri-implantation and mid-gestation Egfr-m1Cwr homozygous phenotypes are actually manifestations of a trophoblast defect that is altered by strain-specific modifiers. This will be accomplished by constructing aggregation chimeras to determine whether EGF-R is essential for differentiation of trophoblast and/or inner cell mass. Continuing the developmental assessment of EGF-R function, experiments are also outlined for determining the requirement for EGF-R during growth and maturation of oocytes as well as during the early pre-implantation stages. These time points could not be addressed by the Egfr-m1Cwr null allele because of the presence of an oocyte-derived pool of EGF-R. Finally, an important aspect for understanding developmental regulation of EGF-R activity is the biological relevance of a truncated form of the receptor that is secreted but lacks kinase activity. The hypothesis is that the secreted receptor modulates full-length receptor activity by competing for ligand. To test this possibility, the secreted receptor will be deleted or overexpressed using gene targeting and transgenic technology.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
7R01HD026722-10
Application #
6182154
Study Section
Human Embryology and Development Subcommittee 1 (HED)
Program Officer
Tasca, Richard J
Project Start
1990-07-01
Project End
2002-06-30
Budget Start
2000-07-01
Budget End
2002-06-30
Support Year
10
Fiscal Year
2000
Total Cost
$302,277
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Genetics
Type
Schools of Medicine
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Chen, B; Bronson, R T; Klaman, L D et al. (2000) Mice mutant for Egfr and Shp2 have defective cardiac semilunar valvulogenesis. Nat Genet 24:296-9
Thomas, J W; LaMantia, C; Magnuson, T (1998) X-ray-induced mutations in mouse embryonic stem cells. Proc Natl Acad Sci U S A 95:1114-9
Chakravarti, S; Magnuson, T; Lass, J H et al. (1998) Lumican regulates collagen fibril assembly: skin fragility and corneal opacity in the absence of lumican. J Cell Biol 141:1277-86
Threadgill, D W; Matin, A; Yee, D et al. (1997) SSLPs to map genetic differences between the 129 inbred strains and closed-colony, random-bred CD-1 mice. Mamm Genome 8:441-2
Threadgill, D W; Yee, D; Matin, A et al. (1997) Genealogy of the 129 inbred strains: 129/SvJ is a contaminated inbred strain. Mamm Genome 8:390-3
Rabinowitz, J E; Rutishauser, U; Magnuson, T (1996) Targeted mutation of Ncam to produce a secreted molecule results in a dominant embryonic lethality. Proc Natl Acad Sci U S A 93:6421-4
Hu, H; Tomasiewicz, H; Magnuson, T et al. (1996) The role of polysialic acid in migration of olfactory bulb interneuron precursors in the subventricular zone. Neuron 16:735-43
Tong, B J; Das, S K; Threadgill, D et al. (1996) Differential expression of the full-length and truncated forms of the epidermal growth factor receptor in the preimplantation mouse uterus and blastocyst. Endocrinology 137:1492-6
Rabinowitz, J E; Magnuson, T (1995) Independent gene targeting by coelectroporation of multiple vectors. Anal Biochem 228:180-2
Threadgill, D W; Dlugosz, A A; Hansen, L A et al. (1995) Targeted disruption of mouse EGF receptor: effect of genetic background on mutant phenotype. Science 269:230-4

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