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Molecular and Cellular Biology, August 2006, p. 6185-6196, Vol. 26, No. 16
0270-7306/06/$08.00+0     doi:10.1128/MCB.00018-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

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

Kojiro Yae,¹ Vincent W. Keng,² Masato Koike,³ Kosuke Yusa,¹ Michiyoshi Kouno,¹ Yoshihiro Uno,⁴ Gen Kondoh,⁵ Takahiro Gotow,⁶ Yasuo Uchiyama,³ Kyoji Horie,¹ and Junji Takeda^1,2*

Departments of Social and Environmental Medicine,¹ Cell Biology and Neurosciences,³ Experimental Animal Science, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka,⁴ Center for Advanced Science and Innovation, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871,² Laboratory of Animal Experiments for Regeneration, Institute for Frontier Medical Science, Kyoto University, 53 Syogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507,⁵ Laboratory of Cell Biology, College of Nutrition, Koshien University, Hyogo 665-0006, Japan⁶

Received 5 January 2006/ Returned for modification 9 March 2006/ Accepted 25 May 2006

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.

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* Corresponding author. Mailing address: Department of Social and Environmental Medicine H3, Osaka University, Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan. Phone: 81-6-6879-3262. Fax: 81-6-6879-3266. E-mail: takeda{at}mr-envi.med.osaka-u.ac.jp.

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Molecular and Cellular Biology, August 2006, p. 6185-6196, Vol. 26, No. 16
0270-7306/06/$08.00+0     doi:10.1128/MCB.00018-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

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