The effect of tartrate on bone cell acid phosphatase activity: a quantitative cytochemical study.
Tartrate-resistant acid phosphatase activity (TRAPase) is widely used as a cytochemical marker to distinguish osteoclasts from macrophages and other related cell types. The degree of tartrate resistance, however, may depend on which reaction methods, tissues, or species are used. To investigate this further, we have measured the amount of cytochemical reaction product by microdensitometry. We compared osteoclast acid phosphatase (APase) activity in fresh frozen sections of neonatal rat calvaria using two different reaction methods; one is commonly employed for qualitative histochemistry and includes alpha naphthyl phosphate as substrate, simultaneous coupling to the chromagen Fast Garnet, and a 30-minute reaction time (method A). The other may be used to measure enzyme reaction rates in cells in situ and employs conditions suitable for initial velocity kinetics, namely naphthol-ASBI phosphate as substrate, post coupling to Fast Garnet, and a 2-minute reaction time. Although enzyme reaction rates differed greatly between the two methods, significant inhibition of APase activity by tartrate was observed in calvarial osteoclasts (69% and 59% with methods A and B, respectively), osteoblasts, and spleen macrophages. Using method B, mouse calvarial osteoclasts had similar APase activity to that seen in the rat. Tartrate produced little inhibition in these mouse cells, in contrast to the observations made with rat tissue, but murine spleen macrophages were significantly tartrate sensitive (40% inhibition with tartrate). On this basis, conclusions regarding the cell specificity of TRAPase should be treated cautiously.
Webber
D
D
University of Manchester Department of Medicine and Endocrinology, Hope Hospital, Eccles, Salford, UK.
Braidman
I P
IP
Robertson
W R
WR
Anderson
D C
DC
D000135:Acid Phosphatase / Q000378:metabolism*; D000818:Animals; D001842:Bone and Bones / Q000187:drug effects / Q000196:embryology / Q000201:enzymology; D003720:Densitometry; D006651:Histocytochemistry; D008264:Macrophages / Q000201:enzymology; D051379:Mice; D010010:Osteoclasts / Q000187:drug effects / Q000201:enzymology*; D051381:Rats; D013154:Spleen / Q000201:enzymology; D013644:Tartrates / Q000494:pharmacology*
D013644:Tartrates; D000135:Acid Phosphatase; C029768:tartaric acid