Immunocytochemical study of cathepsin l and rat salivary cystatin-3 in rat osteoclasts treated with e-64 in vivo

Ryoji Moroi, Takayoshi Yamaza, Toshihiro Nishiura, Yukio Nishimura, Yoshihiro Terada, Kimio Abe, Masaru Himeno, Teruo Tanaka

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Abstract

The localization of cathepsin L and rat salivary cystatin-3 (RSC-3) in rat osteoclasts (rat femoral and alveolar bones) treated with or without E- 64 (control) was examined immunocytochemically. In osteoclasts pretreated with E-64, immunoreactivity for cathepsin L was very weak extracellularly compared to that in the control osteoclasts. However, it was strong intracellularly. The localization of RSC-3 was unclear in the control osteoclasts, while in E-64 treated osteoclasts, both the clear zone and ruffled border areas showed a very strong immunoreaction. At the electron- microscopic level, in normal osteoclasts, numerous immunoreaction products for cathepsin L were found extracellularly in the bone matrix under the ruffled border, while few intracellular products were observed. In contrast, in the E-64-treated osteoclasts, only a few immunoreaction products were found extracellularly, while intracellularly cathepsin L was found in numerous endosome-lysosomal vacuoles. In the immunoreaction for RSC-3, the cytoplasm of the ruffled border was positive, and the tips of the RSC-3- positive ruffled border appeared to enter deeply into the bone matrix. Intracellularly, the granular reaction products of RSC-3 were found in the vacuoles (probably autophagolysosomes). Thus, in E-64-treated osteoclasts, inhibition of the extracellular release of cathepsin L was demonstrated. In addition, intralysosomal accumulation of RSC-3 and deep penetration of the RSC-3-positive ruffled border into the hone matrix were found. These findings suggest that RSC-3 is associated with the inhibition of cathepsin L in both the lysosomes (in the osteoclasts) and bone matrix.

Original languageEnglish
Pages (from-to)305-315
Number of pages11
JournalArchives of Oral Biology
Volume42
Issue number4
DOIs
Publication statusPublished - Apr 1 1997

Fingerprint

Salivary Cystatins
Cystatin C
Cathepsins
Osteoclasts
Cathepsin L
Bone Matrix
Vacuoles
Endosomes
Thigh
Lysosomes
Cytoplasm

All Science Journal Classification (ASJC) codes

  • Otorhinolaryngology
  • Dentistry(all)
  • Cell Biology

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Immunocytochemical study of cathepsin l and rat salivary cystatin-3 in rat osteoclasts treated with e-64 in vivo. / Moroi, Ryoji; Yamaza, Takayoshi; Nishiura, Toshihiro; Nishimura, Yukio; Terada, Yoshihiro; Abe, Kimio; Himeno, Masaru; Tanaka, Teruo.

In: Archives of Oral Biology, Vol. 42, No. 4, 01.04.1997, p. 305-315.

Research output: Contribution to journalArticle

Moroi, Ryoji ; Yamaza, Takayoshi ; Nishiura, Toshihiro ; Nishimura, Yukio ; Terada, Yoshihiro ; Abe, Kimio ; Himeno, Masaru ; Tanaka, Teruo. / Immunocytochemical study of cathepsin l and rat salivary cystatin-3 in rat osteoclasts treated with e-64 in vivo. In: Archives of Oral Biology. 1997 ; Vol. 42, No. 4. pp. 305-315.
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