Expression of typical calpains in mouse molar.
OBJECTIVE
Calpain is a calcium ion-dependent cysteine protease, consisting of two primary isoforms (calpain1/calpain2) which mediate crucial cellular functions. The activity of the calpains is tightly regulated by the endogenous inhibitor calpastatin. Calpains have been detected in several studies during the embryonic and foetal stages. The aim of this study is to investigate the temporal transition of typical calpains and their inhibitor calpastatin during odontogenesis.


METHODS
We used the first molar of foetal ICR mice from embryonic day (E) 14 to postnatal day (PN) 7. Using laser microdissection and semi-quantitative real-time PCR, we investigated calpain1, calpain2 and calpastatin expressions in each enamel epithelium, inner enamel epithelium, stellate reticulum and outer enamel epithelium.


RESULTS
We found calpain1 and calpain2 mRNA increased in the all enamel epithelia between E18 and PN1. In addition calpastatin mRNA expression increased in the ameloblasts from PN1 to PN7. The immunohistochemistry results demonstrated that calpain1/calpain2 was present in the distal side of ameloblasts from PN1 to PN7, and calpastatin was present in the extracellular enamel matrix from E16 to PN1. Furthermore calpain1/calpain2 was present in the dentin, and calpastatin was detected in dentin producing odontoblasts and predentin at PN7.


CONCLUSIONS
In this study the temporal transition of calpain1, calpain2 and calpastatin mRNA and the immunolocalization are identified during tooth development. Our results indicate that the calcium-dependent proteases may play an important role in mouse molar development and extracellular calpain and calpastatin may be involved in molar mineralization.
Matsunaga
T
T
Department of Oral Pathology and Diagnosis, Showa University School of Dentistry, Shinagawa Ward, Tokyo 142-8555, Japan. matsunaga-@dent.showa-u.ac.jp
Yamamoto
G
G
Tachikawa
T
T
D000565:Ameloblasts / Q000378:metabolism*; D000566:Amelogenesis / Q000235:genetics / Q000502:physiology*; D000818:Animals; D002135:Calcium-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D002154:Calpain / Q000096:biosynthesis* / Q000235:genetics; D003743:Dental Enamel / Q000201:enzymology*; D004658:Enamel Organ / Q000196:embryology / Q000201:enzymology*; D004847:Epithelial Cells / Q000378:metabolism; D015870:Gene Expression; D007150:Immunohistochemistry; D051379:Mice; D008813:Mice, Inbred ICR; D008963:Molar / Q000201:enzymology
D002135:Calcium-Binding Proteins; C033668:calpastatin; D002154:Calpain