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Colocalization of APC and DLG at the tips of cellular protrusions in cultured epithelial cells and its dependency on cytoskeletons

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Abstract

Adenomatous polyposis coli gene product (APC) is a tumor suppressor linked to familial adenomatous polyposis and is thought to be involved in cellular polarization and migration in moving epithelial cells. APC interacts with the mammalian homolog of Discs large (DLG). DLG is a member of the membrane-associated guanylate kinase superfamily and is thought to function as a scaffolding protein that coordinates the assembly of a lateral plasma membrane-localized protein complex in epithelial cells. We confirmed the suitability of several anti-APC antibodies for immunocytochemical analysis. Using these antibodies, we showed that APC clusters were colocalized with DLG protein at cellular protrusions of subconfluent MDCK cells. A portion of the clusters was found at the tips of microtubules extending into the cellular protrusions. In addition, actin stress fibers converged near the clusters. When microtubules were disrupted by nocodazole, the colocalization of APC and DLG was lost due to the disappearance of APC clusters. However, the coclusters remained after depolymerization of actin filaments with latrunculin A. This is the first report showing colocalization of APC and DLG in non-polarized epithelial cells. This colocalization suggests that DLG functions not only at the lateral cell–cell contact sites of polarized epithelial cells but also at the protrusions of non-polarized epithelial cells through the interaction with APC protein.

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Acknowledgements

We thank Dr. Tetsu Akiyama from the University of Tokyo for providing anti-DLG antibody (N-ter) and Ms Shiori Fujita for providing technical support. We also thank Dr. Ryuji Nomura, Dr. Yoshimi Hasegawa, and Dr. Hiroshi Kogo for their helpful comments and discussion. This study was supported in part by the OM Award from the Zoological Society of Japan, a Grant-in-Aid for Young Scientists from the Ministry of Education, Culture, Sports, Science and Technology of Japan, a Grant-in-Aid from Fujita Health University, and the Science Research Promotion Fund from the Promotion and Mutual Aid Corporation for Private Schools of Japan.

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  1. Department of Anatomy I, Fujita Health University School of Medicine, Toyoake, 470-1192, Aichi, Japan

    Akiko Iizuka-Kogo, Atsushi Shimomura & Takao Senda

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  1. Akiko Iizuka-Kogo
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  2. Atsushi Shimomura
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  3. Takao Senda
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Correspondence to Takao Senda.

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Iizuka-Kogo, A., Shimomura, A. & Senda, T. Colocalization of APC and DLG at the tips of cellular protrusions in cultured epithelial cells and its dependency on cytoskeletons. Histochem Cell Biol 123, 67–73 (2005). https://doi.org/10.1007/s00418-004-0729-2

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  • DOI: https://doi.org/10.1007/s00418-004-0729-2

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