These Epha2tm1Jrui knock-out mice show impaired ephrin-A1-induced vascular assembly and defective migration, decreased angiogenesis, and failure to activate the Rac1 signaling G protein in response to ephrin-A1.
IMR Colony, The Jackson Laboratory
Mice homozygous for this targeted mutation are viable, fertile, and display no overt developmental or behavioral abnormalities. In mutant mice of a mixed BALB/c, C57BL/6, and 129S6 background, murine pulmonary microvascular endothelial cells (MPMECs) isolated from homozygotes express no endogenous protein. MPMECs from Epha2-deficient mice show impaired ephrin-A1-induced vascular assembly and defective migration both in vitro and in vivo. In addition, MPMECs from homozygous Epha2 mice exhibit decreased angiogenesis and fail to activate Rac1 in response to ephrin-A1 in vivo. Mutant mice on a BALB/c background that were transplanted with metastatic mammary adenocarcinoma cells showed impaired tumor progression, angiogenesis and metastasis to the lung, compared with wildtype littermate controls. These mutant mice may be useful in studies of postnatal angiogenesis (endothelial cell migration, assembly into new tubules, and cytoskeletal regulation), or as a tool to study cancer, especially breast cancer research.
A targeting vector containing the neomycin cassette was used to disrupt exon 5 (at nucleotide 1372, corresponding to amino acid residue 426 in the extracellular domain) of the endogenous Epha2 gene. This construct was microinjected into 129S6/SvEvTac-derived CCE embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. Resulting chimeric males were bred to C57BL/6 females for one generation and backcrossed to BALB/c once before being interbred to homozygosity. Mutant null mice were maintained by sibling matings for more than 10 years prior to arrival at The Jackson Laboratory. The mice were then backcrossed on to the BALB/cByJ (Stock No. 001026) background for 5 generations.
|Allele Name||targeted mutation 1, Joseph C Ruiz|
|Allele Type||Targeted (Null/Knockout)|
|Allele Synonym(s)||Epha2-; Epha2tm1Jinc|
|Gene Symbol and Name||Epha2, Eph receptor A2|
|Gene Synonym(s)||ARCC2; AW545284; CTPA; CTPP1; CTRCT6; ECK; Eck; Eck; Myk2; Sek-2; Sek2; epithelial cell kinase; expressed sequence AW545284|
|Strain of Origin||129S/SvEv-Gpi1 |
|Molecular Note||A neo cassette was inserted at nucleotide 1372, within the sequence encoding the extracellular domain. Although the resultant transcript putatively encodes a nonfunctional protein truncated at amino acid 426, no protein was detected by Western blot analysis of murine pulmonary microvascular endothelial cells from mutant mice.|
|Mutations Made By|| |
Joseph Ruiz, Indiana University School of Medicine
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