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Originally published In Press as doi:10.1074/jbc.C400352200 on August 2, 2004

J. Biol. Chem., Vol. 279, Issue 40, 41263-41266, October 1, 2004
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Defects in Cell Adhesion and the Visceral Endoderm following Ablation of Nonmuscle Myosin Heavy Chain II-A in Mice*{boxs}

Mary Anne Conti{ddagger}§, Sharona Even-Ram||§, Chengyu Liu**, Kenneth M. Yamada||, and Robert S. Adelstein{ddagger}

From the {ddagger}Laboratory of Molecular Cardiology, NHLBI, the ||Craniofacial Developmental Biology and Regeneration Branch, NIDCR, and the **Transgenic Core Facility, NHLBI, National Institutes of Health, Bethesda, Maryland 20892

Previous work has shown that ablation or mutation of nonmuscle myosin heavy chain II-B (NMHC II-B) in mice results in defects in the heart and brain with death occurring between embryonic day 14.5 (E14.5) and birth (Tullio, A. N., Accili, D., Ferrans, V. J., Yu, Z. X., Takeda, K., Grinberg, A., Westphal, H., Preston, Y. A., and Adelstein, R. S. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 12407–12412). Here we show that mice ablated for NMHC II-A fail to develop a normal patterned embryo with a polarized visceral endoderm by E6.5 and die by E7.5. Moreover, A–/A– embryoid bodies grown in suspension culture constantly shed cells. These defects in cell adhesion and tissue organization are explained by loss of E-cadherin and {beta}-catenin localization to cell adhesion sites in both cell culture and in the intact embryos. The defects can be reproduced by introducing siRNA directed against NMHC II-A into wild-type embryonic stem cells. Our results suggest an essential role for a single, specific nonmuscle myosin isoform in maintaining cell-cell adhesions in the early mammalian embryo.

Received for publication, July 26, 2004

* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

{boxs} The on-line version of this article (available at http://www.jbc.org) contains supplemental Experimental Procedures and supplemental Figs. S1–S4.

§ These investigators made major contributions to this work.

To whom correspondence should be addressed: NIH, Bldg. 10, Rm. 8N202, 10 Center Dr., MSC 1762, Bethesda, MD 20892-1762. Tel.: 301-496-1912; Fax: 301-402-1542; E-mail: contim{at}nhlbi.nih.gov.


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