Generation of cortactin floxed mice and cellular analysis of motility in fibroblasts.
Cortactin is an F-actin binding protein that has been suggested to play key roles in various cellular functions. Here, we generated mice carrying floxed alleles of the cortactin (Cttn) gene (Cttn(flox/flox) mice). Expression of Cre recombinase in mouse embryonic fibroblasts (MEFs) isolated from Cttn(flox/flox) embryos depleted cortactin within days, without disturbing F-actin distribution and localization of multiple actin-binding proteins. Cre-mediated deletion of Cttn also did not affect cell migration. To obtain mice with a Cttn null allele, we next crossed Cttn(flox/flox) mice with transgenic mice that express Cre recombinase ubiquitously. Western blot and immunocytochemical analysis confirmed complete elimination of cortactin expression in MEFs carrying homozygously Cttn null alleles. However, we found no marked alteration of F-actin organization and cell migration in Cttn null-MEFs. Thus, our results indicate that depletion of cortactin in MEFs does not profoundly influence actin-dependent cell motility.
Tanaka
Shinji
S
Kunii
Masataka
M
Harada
Akihiro
A
Okabe
Shigeo
S
D000199:Actins / Q000378:metabolism; D000818:Animals; D015139:Blotting, Southern; D002465:Cell Movement / Q000502:physiology*; D051356:Cortactin / Q000235:genetics* / Q000378:metabolism; D003433:Crosses, Genetic; D005347:Fibroblasts / Q000502:physiology*; D018014:Gene Transfer Techniques; D015151:Immunoblotting; D007150:Immunohistochemistry; D019426:Integrases; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic
D000199:Actins; D051356:Cortactin; C045073:Cre recombinase; D019426:Integrases