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MBC in Press, published online ahead of print September 12, 2007
Mol. Biol. Cell 10.1091/mbc.E07-04-0296

A more recent version of this article appeared on November 1, 2007
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Submitted on April 2, 2007
Revised on August 21, 2007
Accepted on August 30, 2007

Amphiphysin 1 Is Important for Actin Polymerization during Phagocytosis

Hiroshi Yamada,* Emiko Ohashi,* Tadashi Abe,* Norihiro Kusumi,{dagger} Shun-AI Li,* Yumi Yoshida,* Masami Watanabe,{dagger} Kazuhito Tomizawa,{ddagger} Yuji Kashiwakura,{sect} Hiromi Kumon,{dagger} Hideki Matsui,{ddagger} and Kohji Takei*

Departments of *Neuroscience, {dagger}Urology, and {ddagger}Cell Physiology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama 700-8558, Japan; {sect}Innovation Center Okayama for Nanobio-Targeted Therapy, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University, 2-5-1 Shikata-cho, Okayama, 700-8558, Japan

Monitoring Editor: David Drubin

Amphiphysin 1 is involved in clathrin-mediated endocytosis. In this study, we demonstrate that amphiphysin 1 is essential for cellular phagocytosis and that it is critical for actin polymerization. Phagocytosis in Sertoli cells was induced by stimulating phosphatidylserine (PS) receptors. This stimulation led to the formation of actin-rich structures, including ruffles, phagocytic cups and phagosomes, all of which showed an accumulation of amphiphysin 1. Knocking out amphiphysin 1 by RNAi in the cells resulted in the reduction of ruffle formation, actin polymerization and phagocytosis. Phagocytosis was also drastically decreased in amph 1 (-/-) Sertoli cells. In addition, phosphatidylinositol 4,5-bisphosphate-induced actin polymerization was decreased in the knockout testis cytosol. The addition of recombinant amphiphysin 1 to the cytosol restored the polymerization process. Ruffle formation in siRNA-treated cells was recovered by the expression of constitutively active Rac1, suggesting that amphiphysin 1 functions upstream of the protein. These findings support that amphiphysin 1 is important in the regulation of actin dynamics and is required for phagocytosis.

Address correspondence to: Kohji Takei (kohji{at}md.okayama-u.ac.jp)




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A. Llobet, M. Wu, and L. Lagnado
The mouth of a dense-core vesicle opens and closes in a concerted action regulated by calcium and amphiphysin
J. Cell Biol., September 22, 2008; 182(5): 1017 - 1028.
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