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Journal of Histochemistry and Cytochemistry
Volume 52 (2): 157-167, 2004
Copyright ©The Histochemical Society, Inc.

Temporal Changes in PTHrP, Bcl-2, Bax, Caspase, TGF-ß, and FGF-2 Expression Following Growth Plate Irradiation With or Without Radioprotectant

Timothy A. Damron, Sharad Mathur, Jason A. Horton, Judith Strauss, Bryan Margulies, William Grant, Cornelia E. Farnum and Joseph A. Spadaro

Departments of Orthopedics and Pathology, SUNY Upstate Medical University (TAD,SM,JAH,JS,BM,JAS), Syracuse, New York, and Cornell University College of Veterinary Medicine (CEF), Ithaca, New York

Correspondence to: Timothy A. Damron, MD, Musculoskeletal Science Research Center, Room 3117, Institute for Human Performance, 505 Irving Ave., Syracuse, NY 13210. E-mail: damront{at}upstate.edu

This study examined temporal changes in growth plate apoptosis molecules and growth factors in an animal model of radiation injury with and without a radioprotectant. Thirty weanling 5-week Sprague–Dawley rats underwent right knee irradiation with single-fraction 17.5 Gy while the left served as internal control. Six animals each were sacrificed at 0.5, 1, 2, 3, or 4 weeks after irradiation. Half of the animals received pretreatment with amifostine (WR-2721) radioprotectant. Immunohistochemical staining for PTHrP, Bcl-2, Bax, caspase-3, FGF-2, and TGF-ß was performed. PTHrP decreased to a nadir at 1 week after irradiation but rebounded to above control levels at 2 weeks in the reserve and transitional zones. The radioprotectant amifostine blunted the decrease in PTHrP but kept PTHrP expression lower than controls during the rebound phase in untreated irradiated animals. Hypertrophic zone Bax expression was decreased by amifostine in both irradiated and non-irradiated limbs at 1 and 2 weeks. FGF, TGF-ß, Bcl-2, and caspase levels generally decreased at 1 week and returned thereafter toward control levels. These findings underscore the importance of PTHrP in response to growth plate irradiation and show the novel finding of a decrease in Bax expression with amifostine pretreatment.

(J Histochem Cytochem 52:157–167, 2004)

Key Words: growth plate • radiation • rat • growth • chondrocyte • radioprotection • apoptosis • amifostine • PTHrP • Bax


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