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Co‐detection of PTH/PTHrP receptor and tartrate resistant acid phosphatase in osteoclasts
Author(s) -
Gay Carol V.,
Zheng Betty,
Gilman Virginia R.
Publication year - 2003
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.10579
Subject(s) - immunostaining , tartrate resistant acid phosphatase , acid phosphatase , receptor , chemistry , antibody , endocrinology , microbiology and biotechnology , medicine , confocal microscopy , tartrate , frozen section procedure , immunohistochemistry , alkaline phosphatase , osteoclast , pathology , biochemistry , biology , immunology , enzyme
Serial sections of rat metaphyses were prepared from paraffin embedded tissue blocks and analyzed in sets of three. The central section was stained for tartrate resistant acid phosphatase (TRAP) in order to identify osteoclasts, one adjacent section was immunostained with an affinity purified antibody to a 15 amino acid sequence unique to rat PTH/PTHrP receptor, and the other adjacent section in the set served as an immunostaining control. This allowed each of the 110 osteoclasts examined to be identified by TRAP and to be tested for the presence or absence of PTH/PTHrP receptor. All antibody solutions and rinses contained 1% donkey serum and 0.5% Tween 20 to ensure antibody integrity and good rinsing procedure. Confocal microscopy was used to evaluate fluorescence intensity of the immunostained osteoclasts. Pixel intensities of 58 osteoclasts from young (4 month) rats and 52 osteoclasts from old (15 month) rats were obtained. Pixel intensities were similar ( P = 0.89) for both young and old animals. However, the number of PTH/PTHrP receptor deficient osteoclasts was greater for the older animals (14.29% vs. 7.24%). This provides direct evidence of PTH/PTHrP receptors in osteoclasts. J. Cell. Biochem. 89: 902–908, 2003. © 2003 Wiley‐Liss, Inc.