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J. Biol. Chem., Vol. 279, Issue 11, 10286-10292, March 12, 2004

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Laminin 2 Is Essential for Odontoblast Differentiation Regulating Dentin Sialoprotein Expression^*

Kenji Yuasa¶, Satoshi Fukumoto¶||, Yoko Kamasaki, Aya Yamada, Emiko Fukumoto**, Kazuhiro Kanaoka, Kan Saito, Hidemitsu Harada, Eri Arikawa-Hirasawa, Yuko Miyagoe-Suzuki¶¶, Shinichi Takeda¶¶, Kuniaki Okamoto, Yuzo Kato, and Taku Fujiwara

From the Division of Pediatric Dentistry and Oral Molecular Pharmacology, Department of Developmental and Reconstructive Medicine, and ^**Division of Oral Health Services Research, Department of Public Health, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki 852-8588, Department of Oral Anatomy and Developmental Biology, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka 565-0871, the Department of Neurology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, 113-8421 Tokyo, and the ^¶¶Department of Molecular Therapy, National Institute of Neuroscience, National Center of Neurology and Psychiatry, 4-1-1 Ogawa-higashi, Kodaira, Tokyo 187-8502, Japan

Laminin 2 is subunit of laminin-2 (211), which is a major component of the muscle basement membrane. Although the laminin 2 chain is expressed in the early stage of dental mesenchyme development and localized in the tooth germ basement membrane, its expression pattern in the late stage of tooth germ development and molecular roles are not clearly understood. We analyzed the role of laminin 2 in tooth development by using targeted mice with a disrupted lama2 gene. Laminin 2 is expressed in dental mesenchymal cells, especially in odontoblasts and during the maturation stage of ameloblasts, but not in the pre-secretory or secretory stages of ameloblasts. Lama2 mutant mice have thin dentin and a widely opened dentinal tube, as compared with wild-type and heterozygote mice, which is similar to the phenotype of dentinogenesis imperfecta. During dentin formation, the expression of dentin sialoprotein, a marker of odontoblast differentiation, was found to be decreased in odontoblasts from mutant mice. Furthermore, in primary cultures of dental mesenchymal cells, dentin matrix protein, and dentin sialophosphoprotein, mRNA expression was increased in laminin-2 coated dishes but not in those coated with other matrices, fibronectin, or type I collagen. Our results suggest that laminin 2 is essential for odontoblast differentiation and regulates the expression of dentin matrix proteins.

Received for publication, September 9, 2003 , and in revised form, December 1, 2003.

^* This work was supported by Grants-in-aid for Scientific Research in a Priority Area 15689025 and 15791255 from the Ministry of Education, Science, Sports and Culture of Japan. 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.

^¶ Both authors contributed equally to this work.

^|| To whom correspondence should be addressed: Division of Pediatric Dentistry, Dept. of Developmental and Reconstructive Medicine, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki 852-8588, Japan. Tel.: 81-95-849-7674; Fax: 81-95-849-7675; E-mail: satoshi{at}dh.nagasaki-u.ac.jp.

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