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Detection of mRNA for carbonic anhydrase II in human osteoclast‐like cells by in situ hybridization
Author(s) -
Zheng Ming H.,
Fan Ying,
Wysocki Stan,
Wood David J.,
Papadimitriou John M.
Publication year - 1993
Publication title -
journal of bone and mineral research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.882
H-Index - 241
eISSN - 1523-4681
pISSN - 0884-0431
DOI - 10.1002/jbmr.5650080114
Subject(s) - in situ hybridization , microbiology and biotechnology , osteoclast , carbonic anhydrase , bone resorption , giant cell tumor of bone , giant cell , carbonic anhydrase ii , northern blot , digoxigenin , chemistry , cytoplasm , carbonic anhydrase i , gene expression , biology , biochemistry , gene , enzyme , in vitro , endocrinology , genetics
Carbonic anhydrase II (CA II) plays an important role during osteoclastic bone resorption. Biochemical investigations of gene expression of CA II, however, have been hampered by difficulty in obtaining sufficient numbers of purified osteoclasts. In this study, we describe a nonradioactive, digoxigenin‐labeled cDNA in situ hybridization technique capable of determining the pattern of CA II gene expression in human osteoclast‐like cells (OC‐like cells) at the single‐cell level. The results showed that CA II mRNA was located in the cytoplasm of both imprinted and cultured OC‐like cells from a giant cell tumor of bone. On the other hand, no evidence of CA II mRNA was found in either the mononuclear cells (tumor cells) of giant cell tumor of bone or osteosarcoma cells. There is a significant correlation between in situ hybridization and northern blot analysis for CA II mRNA in both the giant cell tumor of bone and the osteosarcoma. Our results also indicated that quantitation of in situ hybridization can be achieved by computed cytophotometry.