Sleeping Beauty Transposon-Based Phenotypic Analysis of Mice: Lack of Arpc3 Results in Defective Trophoblast Outgrowth

Abstract

The Sleeping Beauty (SB) transposon system has generated many transposon-insertional mutant mouse lines, some of which have resulted in embryonic lethality when bred to homozygosity. Here we report one such insertion mapped to the mouse actin-related protein complex subunit 3 gene (Arpc3). Arpc3 is a component of the Arp2/3 complex, which plays a major role in actin nucleation with Y-shaped branching from the mother actin filament in response to migration signaling. Arpc3 transposon-inserted mutants developed only to the blastocyst stage. In vitro blastocyst culture of Arpc3 mutants exhibited severe spreading impairment of trophoblasts. This phenotype was also observed in compound heterozygotes generated using conventional gene-targeted and transposon-inserted alleles. Arpc3-deficient mutants were shown to lack actin-rich structures in the spreading trophoblast. Electron microscopic analysis demonstrated the lack of mesh-like structures at the cell periphery, suggesting a role of Arpc3 in Y-shaped branching formation. These data indicate the importance of Arpc3 in the Arp2/3 complex for trophoblast outgrowth and suggest that Arpc3 may be indispensable for implantation.

We thank M. Nozaki for providing the Troma-1 antibody. We are grateful to E. Oiki, R. Ikeda, E. S. Saito, T. Hayakawa, R. Hashimoto, C. Kokubu, H. Fukui, K. Kuratani, K. Yokota, K. Yoshino, M. Yamamoto, M. Okabe, M. Ikawa, and H. Hamada for helpful advice and excellent technical support.

This work was supported by grants from the New Energy and Industrial Technology Development Organization of Japan; the Uehara Memorial Foundation; the Preventure Program, Japan Science and Technology Agency; and RIKEN, The Institute of Physical and Chemical Research. It was also supported by a grant-in-aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.