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Originally published In Press as doi:10.1074/jbc.C100588200 on December 21, 2001

J. Biol. Chem., Vol. 277, Issue 7, 4601-4604, February 15, 2002
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ACCELERATED PUBLICATION
Synaptotagmin IX Regulates Ca2+-dependent Secretion in PC12 Cells*

Mitsunori FukudaDagger §, Judith A. Kowalchyk, Xiaodong Zhang, Thomas F. J. Martin, and Katsuhiko MikoshibaDagger ||

From the Dagger  Laboratory for Developmental Neurobiology, Brain Science Institute, RIKEN (The Institute of Physical and Chemical Research), 2-1 Hirosawa, Wako, Saitama 351-0198, Japan, the  Department of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706, and the || Division of Molecular Neurobiology, Department of Basic Medical Science, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan

Synaptotagmin (Syt) I-deficient phaeochromocytoma (PC12) cell lines show normal Ca2+-dependent norepinephrine (NE) release (Shoji-Kasai, Y., Yoshida, A., Sato, K., Hoshino, T., Ogura, A., Kondo, S., Fujimoto, Y., Kuwahara, R., Kato, R., and Takahashi, M. (1992) Science 256, 1821-1823). To identify an alternative Ca2+ sensor, we searched for other Syt isoforms in Syt I-deficient PC12 cells and identified Syt IX, an isoform closely related to Syt I, as an abundantly expressed dense-core vesicle protein. Here we show that Syt IX is required for the Ca2+-dependent release of NE from PC12 cells. Antibodies directed against the C2A domain of either Syt IX or Syt I inhibited Ca2+-dependent NE release in permeable PC12 cells indicating that both Syt proteins function in dense-core vesicle exocytosis. Our results support the idea that Syt family proteins that co-reside on secretory vesicles may function cooperatively and redundantly as potential Ca2+ sensors for exocytosis.

* This work was supported in part by grants from the Science and Technology Agency to Japan (to K. M.), Grant 13780624 from the Ministry of Education, Science, and Culture of Japan (to M. F.), and National Institutes of Health Grant DK25861 (to T. F. J. M.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ To whom correspondence should be addressed. Tel.: 81-48-467-9745; Fax: 81-48-467-9744; E-mail: mnfukuda@brain.riken.go.jp.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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