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Measuring Calpain Activity in Fixed and Living Cells by Flow Cytometry
Published on: July 8, 2010
Expression of typical calpains in mouse molar
T Matsunaga1, G Yamamoto, T Tachikawa
1Department of Oral Pathology and Diagnosis, Showa University School of Dentistry, Shinagawa Ward, Tokyo 142-8555, Japan. matsunaga-@dent.showa-u.ac.jp
Archives of Oral Biology
|August 18, 2009
Summary
This study reveals that calpain1, calpain2, and calpastatin play key roles during mouse tooth development. Their expression patterns suggest involvement in molar mineralization processes.
Area of Science:
- Biochemistry
- Developmental Biology
- Molecular Biology
Background:
- Calpains (calpain1/calpain2) are calcium-dependent cysteine proteases essential for cellular functions.
- Calpastatin is the endogenous inhibitor regulating calpain activity.
- Calpains are present during embryonic and foetal development.
Purpose of the Study:
- To investigate the temporal expression and localization of calpain1, calpain2, and calpastatin during mouse odontogenesis.
- To understand the role of these proteins in tooth development.
Main Methods:
- Utilized first molar tissues from embryonic day (E) 14 to postnatal day (PN) 7 in ICR mice.
- Employed laser microdissection and semi-quantitative real-time PCR to analyze gene expression.
- Conducted immunohistochemistry to determine protein localization.
Main Results:
- Calpain1 and calpain2 mRNA levels increased in enamel epithelia from E18 to PN1.
- Calpastatin mRNA expression rose in ameloblasts from PN1 to PN7.
- Proteins were localized in ameloblasts, extracellular enamel matrix, dentin, and odontoblasts, with temporal variations.
Conclusions:
- The study identified the temporal expression and localization of calpain1, calpain2, and calpastatin during mouse tooth development.
- Results suggest calcium-dependent proteases are crucial for molar development.
- Extracellular calpain and calpastatin may be involved in molar mineralization.
