An in situ hybridization study of perlecan, DMP1, and MEPE in developing condylar cartilage of the fetal mouse mandible and limb bud cartilage

  • K. Fujikawa Tokyo Medical and Dental University, Japan.
  • T. Yokohama-Tamaki Tokyo Medical and Dental University, Japan.
  • T. Morita Tokyo Medical and Dental Universit, Japan.
  • O. Baba Ohu University - Fukushima, Japan.
  • C. Qin Texas A & M University, United States.
  • S. Shibata | sshibata.mfa@tmd.ac.jp Tokyo Medical and Dental University, Japan.

Abstract

The main purpose of this in situ hybridization study was to investigate mRNA expression of three bone/cartilage matrix components (perlecan, DMP1, and MEPE) in developing primary (tibial) and secondary (condylar) cartilage. Perlecan mRNA expression was first detected in newly formed chondrocytes in tibial cartilage at E13.0, but this expression decreased in hypertrophic chondrocytes at E14.0. In contrast, at E15.0, perlecan mRNA was first detected in the newly formed chondrocytes of condylar cartilage; these chondrocytes had characteristics of hypertrophic chondrocytes, which confirmed the previous observation that progenitor cells of developing secondary cartilage rapidly differentiate into hypertrophic chondrocytes. DMP1 mRNA was detected in many chondrocytes within the lower hypertrophic cell zone in tibial cartilage at E14.0. In contrast, DMP1 mRNA expression was only transiently detected in a few chondrocytes of condylar cartilage at E15.0. Thus, DMP1 may be less important in the developing condylar cartilage than in the tibial cartilage. Another purpose of this study was to test the hypothesis that MEPE may be a useful marker molecule for cartilage. MEPE mRNA was not detected in any chondrocytes in either tibial or condylar cartilage; however, MEPE immunoreactivity was detected throughout the cartilage matrix. Western immunoblot analysis demonstrated that MEPE antibody recognized two bands, one of 67 kDa and another of 59 kDa, in cartilage-derived samples. Thus MEPE protein may gradually accumulate in the cartilage, even though mRNA expression levels were below the limits of detection of in situ hybridization. Ultimately, we could not designate MEPE as a marker molecule for cartilage, and would modify our original hypothesis.

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Author Biographies

K. Fujikawa, Tokyo Medical and Dental University

Department of Maxillofacial Anatomy
Division of Maxillofacial and Neck Reconstruction
Graduate School of Medical and Dental Sciences

T. Yokohama-Tamaki, Tokyo Medical and Dental University

Department of Maxillofacial Anatomy
Division of Maxillofacial and Neck Reconstruction
Graduate School of Medical and Dental Sciences

T. Morita, Tokyo Medical and Dental Universit

Department of Maxillofacial Anatomy
Division of Maxillofacial and Neck Reconstruction
Graduate School of Medical and Dental Sciences

O. Baba, Ohu University - Fukushima
Section of Biology, Department of Oral Function and Molecular Biology, School of Dentistry
C. Qin, Texas A & M University
Department of Biomedical Sciences, Baylor Collage of Dentistry, System Health Science Center
S. Shibata, Tokyo Medical and Dental University

Department of Maxillofacial Anatomy
Division of Maxillofacial and Neck Reconstruction
Graduate School of Medical and Dental Sciences

Published
2015-09-25
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Section
Original Papers
Supporting Agencies
Ministry of Education, Culture, Sports, Science, and Technology of Japan
Keywords:
Mandibular condylar cartilage, perlecan, DMP1, MEPE, in situ hybridization.
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How to Cite
Fujikawa, K., Yokohama-Tamaki, T., Morita, T., Baba, O., Qin, C., & Shibata, S. (2015). An in situ hybridization study of perlecan, DMP1, and MEPE in developing condylar cartilage of the fetal mouse mandible and limb bud cartilage. European Journal of Histochemistry, 59(3). https://doi.org/10.4081/ejh.2015.2553