Cytochemical localization of tartrate‐resistant acid phosphatase, alkaline phosphatase, and nonspecific esterase in perivascular cells of cartilage canals in the developing mouse epiphysis

Abstract

Cytochemical localization of tartrate-resistant acid phosphatase (TRAP), tartrate-sensitive acid phosphatases (TSAP), alkaline phosphatase, and nonspecific esterase was used to characterize perivascular cells within cartilage canals. In the distal femoral epiphyses of 5- to 7-day-old mice, three stages of canal development can be distinguished, and at each developmental stage different perivascular cells were present with morphological characteristics of degradative cells. Vacuolated cells resembling macrophages, fibroblastic cells, and chondroclasts were present adjacent to the matrix in superficial, intermediate, and deep canals, respectively. In order to characterize these perivascular cells cytochemically, nonspecific esterase and TSAP staining was used to identify macrophages, alkaline phosphatase staining was used to identify fibroblastic cells, and TRAP staining was used to identify chondroclasts. There were no cells present in the canals at any developmental stage that were positive for TSAP or strongly positive for nonspecific esterase, placing doubt on the identity of the vacuolated cells as macrophages. Alkaline phosphatase-positive perivascular cells were present in the intermediate and deep canals adjacent to matrix containing alkaline phosphatase-positive chondrocytes. These alkaline phosphatase-positive cells were found in the same location within canals as the fibroblastic cells. Tartrate-resistant acid phosphatase was localized in chondroclasts at the tips of deep canals but was not confined exclusively to chondroclasts. Except for the very early stage of canal development prior to chondrocyte hypertrophy, TRAP-positive cells were present at the tips of superficial and intermediate canals as well as at the tips of the deep canals.(ABSTRACT TRUNCATED AT 250 WORDS)

Growth cartilage calcification and formation of bone trabeculae are late and dissociated events in the endochondral ossification of Rana catesbeiana

“…×900 of endochondral ossification in bullfrogs may represent an economic adaptation for bone growth. The resorption of calcified cartilage and bone by osteoclasts and the resorption of uncalcified cartilage by mononucleated cells are similar to those in birds and mammals (Schenk et al 1967;Cole and Wezeman 1987). AlkPase activity is apparently constitutive in differentiating chondrocytes in bullfrog growth cartilage, with the reaction being detected from the proliferating zone to the hypertrophic zone.…”

“…Alkaline phosphatase (AlkPase) activity was detected by using 0.1% α-naphthyl phosphate, 0.1% Fast red and 50 mM MgCl 2 in 0.1 M TRIS-malate buffer, pH 10. For the identification of osteoclasts, the same compounds were dissolved in 0.1 M acetate buffer, pH 5.0, plus 50 mM sodium tartrate for tartrate-resistant acid phosphatase (TRAP) activity (Minkin 1982;Cole and Wezeman 1987). Controls were prepared by using the same solutions without α-naphthyl phosphate.…”

Ectopic mineralization of articular cartilage in the bullfrog Rana catesbeiana and its possible involvement in bone closure

“…Two-micrometer-thick sections were assayed for alkaline phosphatase (AlkPase) activity using 0.1% α-naphthyl phosphate, 0.1% fast red and 50 mM MgCl 2 in 0.1 M TRIS-malate buffer, pH 10.0. Controls were run in the same solutions without α-naphthyl phosphate (Bancroft 1982;Cole and Wezeman 1987). Sections were counterstained with methyl green and mounted in 80% glycerol.…”

Immunolocalization of receptor activator of nuclear factor‐κB ligand (RANKL) and osteoprotegerin (OPG) in Meckel's cartilage compared with developing endochondral bones in mice

“…During the process of resorption in the anterior midportion, corresponding to the middle portion in the present study, two distinct types of TRAP‐positive cells, chondroclasts and macrophages, can be classified based on their size, number of nuclei and macrophage‐specific antibody (Harada & Ishizeki, 1998). These cells have also been observed during the disintegration of developing mouse epiphyseal cartilage (Cole & Wezeman, 1985, 1987). According to those reports, multinuclear chondroclasts appear to be involved in the removal of cartilaginous matrix and cellular debris.…”